Objective:To explore the cardioprotective effect of hesperidin against arsenic trioxide-induced cardiac toxicity in rats.Methods:Cardiac toxicity was induced by oral administration of 4 mg/kg arsenic trioxide for 30 d...Objective:To explore the cardioprotective effect of hesperidin against arsenic trioxide-induced cardiac toxicity in rats.Methods:Cardiac toxicity was induced by oral administration of 4 mg/kg arsenic trioxide for 30 days.Hematological,biochemical,electrocardiography,echocardiography,and histopathological examinations were performed.Results:Hesperidin decreased the neutrophil-to-lymphocyte ratio,calcium,creatine kinase-myoglobin binding,lactate dehydrogenase,IL-6,and lipid peroxidation,as well as increased sodium and potassium concentration and superoxide dismutase and catalase activity in arsenic trioxide-intoxicated rats.Moreover,it reduced peak systolic velocity and end-diastolic velocity while increasing heart rate.Arsenic trioxide-induced histopathological damage to cardiac tissue was prominently alleviated by hesperidin treatment.Conclusions:Hesperidin attenuates arsenic trioxide-induced cardiac toxicity in rats.Therefore,it can be further explored as a cardioprotective agent.展开更多
OBJECTIVE To study the anti-tumor effect of arsenic trioxide on the HepG2 human hepatocellular carcinoma cell line, and to explore its mechanism of action. METHODS The MTT assay was used to determine the inhibitory ef...OBJECTIVE To study the anti-tumor effect of arsenic trioxide on the HepG2 human hepatocellular carcinoma cell line, and to explore its mechanism of action. METHODS The MTT assay was used to determine the inhibitory effect of As2O3 on HepG2 cells at various As2O3 concentrations. The expression of p-JNK, caspase-3 and PARP was detected by Western blots. RESULTS As2O3 markedly inhibited the growth of the HepG2 cells and induced apoptosis. The results of Western blot analysis showed that the As2O3-induced apoptosis was accompanied by caspase-3 and PARP activation. p-JNK was detected at 10 min following As2O3 treatment, and preceded to peak at 20 min, and decreased by 30 min. The total protein content did not obviously change. The activation of JNK occurred prior to cell apoptosis. SP600125, a JNK inhibitor, suppressed the As2O3-induced activation of caspase-3 and PARP cleavage. CONCLUSION As2O3 inhibits the proliferation of human HepG2 hepatocellular carcinoma cells by inducing apoptosis in vitro. As2O3-induced apoptosis is accessed through the caspase-3 pathway. The JNK signal-transduction pathway and caspase-3 are involved upstream in the As2O3 induced HepG2 apoptotic response.展开更多
AIM To study the effect of a varyingconcentrations of arsenic trioxide on humanhepatoma cell line BEL-?402 cultured in vitro andits mechanism of action.METHODS The BEL-7402 cells were treatedwith arsenic trioxide（at ...AIM To study the effect of a varyingconcentrations of arsenic trioxide on humanhepatoma cell line BEL-?402 cultured in vitro andits mechanism of action.METHODS The BEL-7402 cells were treatedwith arsenic trioxide（at the concentrations of0.5,1,2 μmol/L,respectively）for 4 successivedays.The cell growth and proliferation wereobserved by cell counting and cell-growth curve.Morphologic changes were studied withelectronmicroscopy.Flow cytometry was usedto assay celI-DNA distribution and the proteinexpression of Bcl-2 and Bax detected byimmunocytochemical method.RESULTS The cell growth was significantlyinhibited by varying concentrations of arsenictrioxide as revealed by cell counting and cell-growth curve,which was dose- and time-dependent.Arsenic trioxide treatment at 0.5,1and 2 μmol/L resulted in a sub-G1 cell peak,theapoptosis rate of the control group was 9.31%and that of 0.5 μmol/L arsenic trioxide 15.53%,no significant difference was seen between thetwo.The apoptosis rates of 1,2 μmol/L arsenictrioxide were 19.10% and 21.87% respectively,which were much higher（both P【0.05）.Decrease of G<sub>0</sub>/G<sub>1</sub> phase cells and increase of Sphase cells were observed by flow cytometry,suggesting the inhibition effect of 0.5,1,2 μmol/L arsenic trioxide on BEL-7402 cell lay in the G<sub>0</sub>/G<sub>1</sub> phase.Morphologic changes such asintact cell membrane,nucleic condensation,apoptotic body formation were seen undertransmission electronmicrescopy,whereas the0.5 mol/L arsenic trioxide-treated BEL-7402cells showed decrease of nucleocytoplasmicratio,round nucleus,well-differentiatedorganelles in the cytoplasm.The processes andmicrovilli on the cell surface of the experimentalgroups under scanning electron microscopy weresignificantly decreased.High expressions ofBcl-2 and Bax were detected in 1 and 2 μmol/Larsenic trioxide-treated cells,these were 46%,87.33% and 83.08%,95.83% respectively,among which that of Bax was more significant.Arsenic trioxide treatment at 0.5 μmol/Lresulted in a higher expression level of Bcl-2 andlower expression level of Bax,which were8.81% and 3.83% respectively,as comparedwith that of the control group（15.33%）（P<sub>1</sub>【0.01,P<sub>2</sub>【0.01）.CONCLUSION Arsenic trioxide not onlyinhibited proliferation but also induced apoptosisof human hepatoma cell line BEL-7402.Theinduced-apoptosis effect of 1,2 μmol/L arsenictrioxide was related to the expression level ofBcl-2 and Bax.展开更多
AIM： To study the anti-hepatoma efficiency of arsenic trioxide （As2O3） in the treatment of experimental rat hepatocellular carcinoma （HCC） induced by 2-acetamidofluorene （2-FAA） and to elucidate the possible me...AIM： To study the anti-hepatoma efficiency of arsenic trioxide （As2O3） in the treatment of experimental rat hepatocellular carcinoma （HCC） induced by 2-acetamidofluorene （2-FAA） and to elucidate the possible mechanisms. METHODS： SD rats （2 mo old） had been fed with 2-FAA for 8 wk to induce HCC, and then they were treated with As2O3 or matrine. On d 29, the rats were killed and the liver was weighed and liver tumors were counted. The histological changes of liver tissue were observed under microscope, and the cellular dynamic parameters were studied by flow cytometry. Immunohistochemistry （two-step method） was used to observe the expression of vascular endothelial growth factor （VEGF） and micro-vessel density （MVD） on consecutive sections. The pathological parameters were also analyzed, the levels of serum aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, total bilirubin （TBi）, and direct bilirubin （DBi）. RESULTS： The number of liver tumors decreased significantly in groups treated with As2O3, especially in medium-dose （1 mg/kg） group （t = 2.80, P〈0.01）. As2O3 caused HCC cell death via apoptosis; necrosis was seen and apoptosis was common when the dose was 1 mg/kg. Proliferation index decreased sharply in medium-dose （1 mg/kg） group （7.87±4.11 vs24.46±6.49, t= 2087, P〈0.01）, but not in 0.2 mg/kg group. However, S-phase fraction decreased dramatically in both groups, it reached the bottom level only when the dose was i mg/kg compared with control （0.40±0.13 vs3.01±0.51, t= 2.97, P〈0.01）, and it was obviously accompanied with accumulation of cells in G0/G1 （G0/G1 restriction）. The expressions of VEGF and MVD in medium-dose （1 mg/kg） group were significantly lower than normal saline group （0.63±0.74 vs2.44±0.88, P〈0.05; 15.75±3.99 vs47.44±13.41, t= 2.80, P〈0.01）. Compared with normal saline group, mediumand low-dose groups As203 and matrine lowered the levels of ALT in serum （61.46±9.46, 63.75±20.40, 61.18±13.00 vs 108.98±29.86, t= 2.14, P〈0.05）, but had no effect onthe level of serum AST, TBi, and DBi. CONCLUSION： As203 had inhibitory effect on growth of experimental HCC in rats induced by 2-FAA, but had no obvious effect on normal hepatic cells. The mechanisms may involve decrease of cell division, accumulation of cells in G0/G1 phase, apoptosis of tumor cells, and inhibitory effect on angiogenesis through blocking VEGF.展开更多
AIM： To investigate the effect of all-trans-retinoic acid （ATRA） on arsenic trioxide （As2O3）-induced apoptosis of human hepatoma, breast cancer, and lung cancer cells in an attempt to find a better combination th...AIM： To investigate the effect of all-trans-retinoic acid （ATRA） on arsenic trioxide （As2O3）-induced apoptosis of human hepatoma, breast cancer, and lung cancer cells in an attempt to find a better combination therapy for solid tumors. METHODS： Human hepatoma cell lines HepG2, Hep3B, human breast cancer cell line MCF-7, and human lung adenocarcinoma cell line AGZY-83-a were treated with As203 together with ATRA. Cell survival fraction was determined by MTT assay, cell viability and apoptosis were measured by annexin V-fluorescein isothiocyanate （FITC） and PI staining, and intracellular glutathione （GSH） and glutathione-S-transferase （GST） activities were determined using commercial kits. RESULTS： Cytotoxicity of ATRA was low. ATRA （0.1, 1, and 10 μmol/L） could synergistically potentiate As2O3 to exert a dose-dependent inhibition of growth and to induce apoptosis in each of the cell lines. HepG2 and Hep3B with low intracellular GSH or GST activities were remarkably sensitive to As2O3 or As2O3＋ATRA, while AGZY-83-a with higher GSH or GST activities was less sensitive to As2O3 or As2O3＋ATRA. Treatment with 2 μmol/L As2O3 for 72 h significantly decreased intracellular GSH and GST levels in each of the cell lines, and 1 μmol/L ATRA alone reduced minimal intracellular GSH and GST levels. ATRA potentiated the effect of As2O3 on intracellular GSH levels, but intracellular GST levels were not significantly affected by the combination of As2O3 and ATRA for 72 h as compared to As2O3 alone.CONCLUSION： ATRA can strongly potentiate As2O3- induced growth-inhibition and apoptosis in each of the cell lines, and two drugs can produce a significant synergic effect. The sensitivity to As2O3 or As2O3＋ATRA is inversely proportional to intracellular GSH or GST levels in each of the cell lines. The GSH redox system may be the possible mechanism by which ATRA synergistically potentiates As203 to exert a dose-dependent inhibition of growth and to induce apoptosis. 2005 The WJG Press and Elsevier Inc. All rights reserved.展开更多
BACKGROUND:It has been pointed out that only low-dose arsenic trioxide(ATO)presents therapeutic benefits outweighing the toxic side effects.Low-dose ATO can effectively alleviate acute promyelocytic leukemia(APL). How...BACKGROUND:It has been pointed out that only low-dose arsenic trioxide(ATO)presents therapeutic benefits outweighing the toxic side effects.Low-dose ATO can effectively alleviate acute promyelocytic leukemia(APL). However,it is quite challenging in treating solid tumors. The purpose of this study was to investigate the effect of ATO at low concentrations on the metastatic potential of mouse hepatoma H22 cells and the anti-metastatic mechanism of ATO. METHODS:The metastatic potential of H22 cells was evaluated by adhesion,migration and invasion assays after exposure to a low dose of ATO in vitro.The mouse lung metastatic model induced by injection of H22 cells via the tail vein was adopted for the evaluation of metastatic potential. Different proteins in the lysate of H22 cells exposed to ATO at different concentrations were investigated by surface- enhanced laser desorption and ionization time-of-flight mass spectrometry(SELDI-TOF-MS).Finally,Western blotting analyses were made to detect the expression pattern of MMP-2 and nm23-M1 proteins. RESULTS:Significant cell death started at ATO concentrations above 2μmol/L.The growth and adhesion potential of H22 cells was inhibited in a time-and dose- dependent manner,and the migration and invasion potential of H22 cells was inhibited in a dose-dependent manner while ATO concentration was below 2μmol/L. Mice injected with ATO at a dose of 0.5 mg/kg had fewer lung metastases.However,mice injected with ATO at a dose of 2 mg/kg or 4 mg/kg had a high mortality rate and more liver injuries.A total of 15 different protein peaks were identified between the lysate of H22 cells treated with ATO and controls.Two proteins that peaked atm/z 5302 and 17207 coincided with MMP-2(fragment) and nm23-M1,respectively.Western blotting analyses demonstrated that MMP-2 and MMP-2 fragments were down-regulated and nm23-M1 was up-regulated in H22 cells treated with 2μmol/L ATO for 48 hours. CONCLUSIONS:ATO at a low dose inhibits the metastatic potential of mouse hepatoma H22 cells in vitro and in vivo, and involves down-regulation of MMP-2 and up-regulation of nm23-M1.展开更多
Objective To reduce the toxicity and side effects of arsenic trioxide(ATO)and provide a new approach for the treatment of primary liver cancer,a folic acid-modified calcium arsenite liposomal“target-controlled”drug ...Objective To reduce the toxicity and side effects of arsenic trioxide(ATO)and provide a new approach for the treatment of primary liver cancer,a folic acid-modified calcium arsenite liposomal“target-controlled”drug delivery system(FA-LP-CaAs)was fabricated using the reverse microemulsion method.Methods A Malvern particle size analyzer and a transmission electron microscope were employed to determine the particle size,distribution,zeta potential and morphology of FA-LP-CaAs.Further,inductively coupled plasma emission spectrometry was employed to determine the drug loading capacity,entrapment efficiency,and in vitro release behavior of FA-LP-CaAs.To determine its toxicity in human hepatoma cells(HepG2)and human normal hepatocytes(LO2)and its effect on HepG2 cell cycle and apoptosis,the MTT method was used.Laser confocal and flow cytometry were also employed to determine the uptake of FA-LP-CaAs by cells.After establishing a mouse liver cancer model,the in vivo distribution of the drug included in the formulation was investigated using in vivo fluorescence.To evaluate the liver cancer targeting and anti-tumor effects of FALP-CaAs in vivo,the distribution of ATO in tissues and changes in tumor volume and body weight after liposomal administration were investigated using hematoxylin-eosin(HE)-stained tumor sections.Results The particle size,zeta potential and PDI of FA-LP-CaAs were(122.67±2.18)nm,(12.81±0.75)mV and 0.22±0.01,respectively,while its drug loading capacity was 18.49%±1.14%.In vitro experimental results revealed that FA-LP-CaAs had a strong killing effect on HepG2 cells.Further,the cell uptake capacity of this formulation was found to improve.Based on in vivo assessments,FA-LP-CaAs could significantly increase the distribution of ATO in tumor sites and inhibit tumor growth.Conclusions Herein,an FA-LP-CaAs formulation was successfully fabricated.This liposomal drug delivery system had a round appearance,uniform particle size,good polydispersity coefficient,evident“core-shell”structure,high drug loading capacity and pH response,tumor targeted drug delivery and sustained drug release.These findings support further research and the application of ATO as an anti-liver cancer prodrug and provide a new method for the treatment of liver cancer.展开更多
OBJECTIVE To explore the inhibitory effect of arsenic trioxide on growth and telomerase activity of BEL-7402 and SMMC-7721 hepatocarcinoma cells, and to measure their GSH level. METHODS Cell culture and trypan blue ex...OBJECTIVE To explore the inhibitory effect of arsenic trioxide on growth and telomerase activity of BEL-7402 and SMMC-7721 hepatocarcinoma cells, and to measure their GSH level. METHODS Cell culture and trypan blue exclusion were used to examine the inhibitory effect of arsenic trioxide on BEL-7402 and SMMC-7721 hepatocarcinoma lines. A GSH kit and telomerase kit were used to mearsure the GSH content in cells and telomerase activity. RESULTS The growth of BEL-7402 cells was significantly inhibited at a level of 0.50μmol/L of arsenic trioxide by 24 h. The inhibitory effect increased with time and concetration of arsenic trioxide. The telomerase activity of BEL-7402 cells was also significantly inhibited at a level of 0.50μmol/L of arsenic trioxide by 24 h, after which the inhibitory effect increased with time. On the other hand, at 24 h of incubation a level of 2.00 μmol/L of arsenic trioxide was required to significantly inhibit growth of SMMC-7721 cells, and only after 48 h with 2.00μmol/L of arsenic trioxide did telomerase activity significantly decline. The GSH content of the BEL-7402 and SMMC-7721 cells was 18.7±1.4 and 50.8±5.2 nmol/mg protein respectively, a significant difference. CONCLUSION Different concentrations of arsenic trioxide are required to inhibit growth and telomerase activity of SMMC-7721 and BEL-7402 cells. Perhaps BEL-7402 cells are more sensitive to arsenic trioxide because of their low level of GSH content, which results in a low capacity for oxidation-reduction and poorer detoxification mechanisms in BEL-7402 cells.展开更多
Objective: The study aimed to investigate the correlation between concentration of inhaled arsenic trioxide and dynamic changes in hematotoxicity in rats. Method: Wistar rats were randomly divided into four study grou...Objective: The study aimed to investigate the correlation between concentration of inhaled arsenic trioxide and dynamic changes in hematotoxicity in rats. Method: Wistar rats were randomly divided into four study groups that were treated with saline(control) or arsenic trioxide at a low(0.1 mg/mL), medium(1 mg/mL), or high(10 mg/mL) dose by intratracheal instillation. Blood samples were collected for analysis at 6, 12, 24, 48, and 72 h after exposure. Results: Compared with the control group, the red blood cell distribution width decreased significantly in each exposure group and then gradually recovered. The red blood cell count only increased in the high-dose group, and the hemoglobin levels did not differ significantly between the groups. Significant decreases in the platelet count and mean platelet volume were observed in each arsenic-exposed group. White blood cell countsalso decreased significantly and then gradually recovered; this was associated with an increased percentage of lymphocytes and a decreased percentage of neutrophils. The blood cell parameter changes were related to the arsenic trioxide concentration and the observation time. Conclusion: Intratracheal instillation of arsenic trioxide affected hematopoietic differentiation in rats, leading to blood cell changes that were related to observation time and concentration.展开更多
Objective： To explore the effects of arsenic trioxide （ATO） on the apoptosis of glucocorticoid （GC）-resistant T-acute lymphoblastic leukemia （ALL） CEM-C1 cells and its possible mechanisms. Methods： Different ...Objective： To explore the effects of arsenic trioxide （ATO） on the apoptosis of glucocorticoid （GC）-resistant T-acute lymphoblastic leukemia （ALL） CEM-C1 cells and its possible mechanisms. Methods： Different concentrations of ATO （0.25 μmol/L-5 μmol/L） were used to induce the apoptosis of CEM-C1 cells. The inhibition rate of cell proliferation and apoptosis were detected by MTT test, Annexin V-FITC/PI flow cytometry and optical microscopy, respectively. RT-PCR was applied to semi-quantitatively analyze the mRNA expression of pro-apoptotic proteins （Bad and PDCD4） and anti-apoptotic proteins （XIAP and MCL-1） induced by different concentrations of ATO at different time points. Results： ATO could inhibit proliferation and induce apoptosis of CEM-C1 cells at a concentration and time dependent manner. Low-dose ATO mildly inhibited the proliferation of CEM-C1 cells while higher concentrations （1 μmol/L and 5 μmol/L） had strong anti-tumor effect with the inhibiting rates of 40.07±7.98% and 88.67±2.88%, respectively. Annexin V-FITC/PI flow cytometry showed that the apoptotic rates of CEM-C1 ceils were significantly increased after 48 hours treatment of different concentrations of ATO. RT-PCR demonstrated up-regulated mRNA expression of pro-apoptotic protein Bad and PDCD4 but down-regulated mRNA expression of anti-apoptotic protein XIAP when CEM-C1 cells were treated with different concentrations of ATO at different time points. The MCL-1 mRNA expression was down-regulated only after the treatment of 5 μmol/L ATO. Conclusion： ATO can inhibit cell proliferation and induce cell apoptosis in GC-resistant CEM-C1 cells. The molecular mechanisms might involve the increased mRNA expression of pro-apoptotic protein Bad and PDCD-4, and rapid down-regulation of XIAP mRNA expression.展开更多
Aim This study was to evaluate the effect of arsenic trioxide （As2O3） on the transgenic TNF-α promoter activity in cultured vascular smooth muscle cells （VSMCs） and THP-1 monocytes. Methods Human TNF-α promoter ...Aim This study was to evaluate the effect of arsenic trioxide （As2O3） on the transgenic TNF-α promoter activity in cultured vascular smooth muscle cells （VSMCs） and THP-1 monocytes. Methods Human TNF-α promoter was constructed by reporter gene system and was transiently transfected into VSMCs and THP-1 in vitro. The promoter activity was tested by luciferase activity with or without LPS and Ang Ⅱ stimulation, before and after different dosage of As2O3 treatment. Results 1. TNF-α promoter effectively expressed in VSMCs and THP-1 compared with CMV promoter （58.3% and 80.9%, respectively）. Both LPS and Ang Ⅱ significantly up-regulated TNF-α promoter activity （P〈0.05）. 2. As2O3 significantly inhibited, both intact and LPS/Ang Ⅱ stimulated promoter activity, in a dose dependent manner （P〈0.05）, and in both cell type. Conclusion These results manifested that, the inhibition effect of As2O3 on the activity of human TNF-α promoter indicated its potential inhibition on pro-inflammatory cytokine genes expression at transcriptional level and its potential anti-inflammatory property in the cardiovascular system.展开更多
Roles of Marigold extracts (ME) on arsenic trioxide (ATO)-induced oxidative damage to pancreatic β-cells need to be further elucidated. In this study, NIT-1 cells were treated with different concentrations of and/or ...Roles of Marigold extracts (ME) on arsenic trioxide (ATO)-induced oxidative damage to pancreatic β-cells need to be further elucidated. In this study, NIT-1 cells were treated with different concentrations of and/or ATO, following by the cell viability was detected by CCK8 assay. Then, intracellular reactive oxygen species (ROS) levels, lipid peroxide (MDA) contents and superoxide dismutase (SOD) activity were measured with a fluorescence probe method and colorimetric assay, respectively. The apoptosis rate and morphology was detected and observed with hoechst 33,258 staining assay. The mRNA levels and protein expressions of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were measured by real-time fluorescence quantitative polymerase chain reaction and protein immunoblotting assay, respectively. Our results indicated that Co-treatment with ME and ATO exacerbated the cell viability decreasing reduced by ATO, while the addition of ME after ATO treatment effectively promote the recovery of ATO reduced survival rates. The ATO group increased apoptosis (P P β-cells by modulating the activation of the Nrf2 signaling pathway.展开更多
AIM To study the effects of arsenic trioxide andHCPT on different degrees of differentiated gastriccancer cells（SGC-7901,MKN-45,MKN-28）withrespect to both cytotoxicity and induction ofapoptosis in vitro.METHODS The ...AIM To study the effects of arsenic trioxide andHCPT on different degrees of differentiated gastriccancer cells（SGC-7901,MKN-45,MKN-28）withrespect to both cytotoxicity and induction ofapoptosis in vitro.METHODS The cytotoxicity of As<sub>2</sub>O<sub>3</sub> and HCPTon gastric cancer cells was determined by MTTassay.Morphologic changes of apoptosis ofgastric cancer cells were observed by lightmicroscopy and transmission electron microscopy.Apoptosis and cell cycle changes of gastric cancercells induced by HCPT and As<sub>2</sub>O<sub>3</sub> were investigatedby TUNEL method and flow cytometry.RESULTS As<sub>2</sub>O<sub>3</sub> and HCPT had remarkablecytotoxic effects on different degrees ofdifferentiated gastric cancer cells.The IC<sub>50</sub>ofAs<sub>2</sub>O<sub>3</sub> on well differentiated gastric cancer cellMKN-28,moderately differentiated gastric cancercell SGC-7901,and poorly differentiated gastriccancer cell MKN-28 were 8.91 μmol/L,10.57μmol/L,and 11.65 μmol/L,respectively.The IC<sub>50</sub>of HCPT on MKN-28,SGC-7901,and MKN-45 were9.35 mg/L,10.21 mg/L,and 12.63 mg/Lrespectively after 48 h treatment.After 12 h ofexposure to both drugs,gastric cancer cellsexhibited morphologic features of apoptosis,including cell shrinkage,nuclear condensation, and formation of apoptotic bodies.A typicalsubdiploid peak before G<sub>0</sub>/G<sub>1</sub> phase was observedby flow cytometry.The apoptotic rates of SGC-7901,MKN-45,and MKN-28 were 13.84%,22.52%,and 9.68%,respectively after 48 hexposure to 10 μmol/L As<sub>2</sub>O<sub>3</sub>.The apoptotic ratesof SGC-7901,MKN-45,and MKN-28 were 21.88%,12.35%,and 30.26%,respectively after 48 hexposure to 10 mg/L HCPT.The apoptotic indicewere 7%-15% as assessed by TUNEL method.The effect of As<sub>2</sub>O<sub>3</sub> on SGC-7901 showedremarkable cell cycle specificity,which inducedcell death in G<sub>1</sub> phase,and blocked G<sub>2</sub>/M phase.HCPT also showed a remarkable cell cyclespecificity,by inducing cell death and apoptosis inG<sub>1</sub> phase and arrest of proliferation at S phase.CONCLUSION As<sub>2</sub>O<sub>3</sub> and HCPT exhibitsignificant cytotoxicity on gastric cancer cells byinduction of apoptosis.As<sub>2</sub>O<sub>3</sub> and HCPT mighthave a promising prospect in the treatment ofgastric cancer,which needs to be further studied.展开更多
Since arsenic trioxide was first approved as the front line therapy for acute promyelocytic leukemia 25 years ago,its anti-cancer properties for various malignancies have been under intense investigation.However,the c...Since arsenic trioxide was first approved as the front line therapy for acute promyelocytic leukemia 25 years ago,its anti-cancer properties for various malignancies have been under intense investigation.However,the clinical successes of arsenic trioxide in treating hematological cancers have not been translated to solid cancers.This is due to arsenic＇s rapid clearance by the body＇s immune system before reaching the tumor site.Several attempts have henceforth been made to increase its bioavailability toward solid cancers without increasing its dosage albeit without much success.This review summarizes the past and current utilization of arsenic trioxide in the medical field with primary focus on the implementation of nanotechnology for arsenic trioxide delivery to solid cancer cells.Different approaches that have been employed to increase arsenic＇s efficacy,specificity and bioavailability to solid cancer cells were evaluated and compared.The potential of combining different approaches or tailoring delivery vehicles to target specific types of solid cancers according to individual cancer characteristics and arsenic chemistry is proposed and discussed.展开更多
AIM: To investigate the effect of arsenic trioxide on human gastric cancer cell line MKN45 with respect to both cytotoxicity and induction of apoptosis in vitro. METHODS: MKN45 cells were treated with arsenic trioxide...AIM: To investigate the effect of arsenic trioxide on human gastric cancer cell line MKN45 with respect to both cytotoxicity and induction of apoptosis in vitro. METHODS: MKN45 cells were treated with arsenic trioxide (As2O3) at the concentration of 1, 5, and 10 μmol/L, respectively, for three successive days. Cell growth and proliferation were observed by cell counting and trypan blue exclusion. Cytotoxicity of As2O3 was determined by MTT assay. Morphologic changes were studied with light microscopy. Flow cytometry was used to assay cell DNA distribution and apoptotic cells were confirmed with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and DNA electrophoresis. RESULTS: The growth of MKN45 cells was significantly inhibited by As2O3 which was confirmed by colony-forming assay. After 7 d of culture with various concentrations of As2O3, colony-forming capacity of MKN45 cells decreased with As2O3 increment in comparison with that of control group. The inhibitory rate of colony-formation was 38.5%, 99.1%, and 99.5% when the concentration of As2O3 was 1, 5, and 10 μmol/L in culture medium, respectively. The cell number of a single colony in drug treatment groups was less than that of control group. The cell-killing rate of As2O3 to MKN45 cells was both dose- and time-dependent with an IC50 of (11.05±0.25) μmol/L After incubation in 10 μmol/L As2O3 for 24 h, the cell-killing rate was 27.1%, and it was close to 50% after 48 h. The results showed that As2O3 induced time- and dose-dependent apoptosis in MKN45 cells, blocked at G2/M phase. The apoptotic peak (sub-G1 phase) appeared and cell apoptotic rate in MKN45 cells was 18.3-32.5% after treatment by 10 umol/L As2O3 for 48 h. The percentage of G2/M cell of the experimental groups was 2.0-5.0 times than that of the control group. Gel electrophoresis of DNA from cells treated with each concentration of As2O3 for 48 h revealed a 'ladder' pattern, indicating preferential DNA degradation at the internucleosomal, linker DNA sections. TUNEL also demonstrated strand breaks in DNA of MKN45 cells treated with As2O3, while control cells showed negative labeling. CONCLUSION: As2O3 can induce apoptosis of human gastric carcinoma cells MKN45, which is the basis of its effectiveness. It shows great potential in the treatment of gastric carcinoma.展开更多
AIM: To investigate the changes in apoptosis in gastrointestinal cancer cells from patients with gastrointestinal cancers treated with arsenic trioxide (As<sub>2</sub>O<sub>3</sub>); and to stu...AIM: To investigate the changes in apoptosis in gastrointestinal cancer cells from patients with gastrointestinal cancers treated with arsenic trioxide (As<sub>2</sub>O<sub>3</sub>); and to study the possible molecular mechanisms of such changes by detecting the expression levels of p53 and Bcl-2.展开更多
AIM: To investigate the inhibitory effect of As2O3 on angiogenesis of tumor and expression of vascular endothelial growth factor (VEGF) in tumor cells in vivo and in vitro. METHODS: The solid tumor model was formed in...AIM: To investigate the inhibitory effect of As2O3 on angiogenesis of tumor and expression of vascular endothelial growth factor (VEGF) in tumor cells in vivo and in vitro. METHODS: The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were randomly divided into three groups. As2O3 was injected into the arsenic-treated groups (2.5 mg/kg and 5 mg/kg) and the same volume of saline solution was injected into the control group. Microvessel density (MVD) and expression of VEGF were detected with immunofluorescence laser confocal technology. Further expression of VEGF protein and VEGF mRNA was measured with Western bloting and fluorescence quantitative RT- PCR in SGC-7901 cells treated with As2O3. RESULTS: In nude mice, after treatment with 5 mg/kg and 2.5 mg/kg As2O3 respectively, about 50% and 30% tumor growth inhibition were observed correspondingly (P < 0.05, P < 0.05). Decrease in MVD appeared in As2O3-treated tumors compared with control group (P < 0.001, P < 0.001). MVD in tumors was significantly lower in 5 mg/kg group than in 2.5 mg/kg group (P < 0.01). The fluorescence intensity levels of VEGF in tumor cells were significantly lowered in the arsenic-treated groups (P < 0.01, P < 0.01). The fluorescence intensity level of VEGF in 5 mg/kg group was lower than that in 2.5 mg/ kg group (P < 0.01). In vitro, the expression of VEGF protein decreased in dose- and time-dependent manner after the treatment with As2O3, but in VEGF mRNA no significant difference was found between the control group and the treated groups. CONCLUSION: As2O3 can inhibit solid tumor growth by inhibiting the formation of new blood vessels. One of the mechanisms is that As2O3 can inhibit VEGF protein expression.展开更多
INTRODUCTION Cell apoptosis,which involves the biologic regulation of the numbers and vital activity of cells,is an important metaboloc process in both normal cells and tumor cells.
AIM： To investigate the effect of arsenic trioxide （As2O3） on expression of vascular endothelial growth factor receptor-1 （VEGFR-1, Flt-1） and VEGFR-2 （KDR） in human gastric tumor cells and proliferation of vas...AIM： To investigate the effect of arsenic trioxide （As2O3） on expression of vascular endothelial growth factor receptor-1 （VEGFR-1, Flt-1） and VEGFR-2 （KDR） in human gastric tumor cells and proliferation of vascular endothelial cells.METHODS： The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were treated with As2O3. Microvessel density （MVD） and expression of Flt-1 and KDR were detected by immunofluorescence laser confocal microscopy. SGC-7901 cells were treated respectively by exogenous recombinant human VEGF165 or VEGF165 ＋ As2O3. Cell viability was measured by MTT assay. Cell viability of ECV304 cells was measured by MTT assay, and cell cycle and apoptosis were analyzed using flow cytometry.RESULTS： The tumor growth inhibition was 30.33% and 50.85%, respectively, in mice treated with As2O3 2.5 and 5 mg/kg. MVD was significantly lower in arsenic-treated mice than in the control group. The fluorescence intensity levels of Flt-1 and KDR were significantly less in the arsenic-treated mice than in the control group. VEGF165 may accelerate growth of SGC7901 cells, but As2O3 may disturb the stimulating effect of VEGF165. ECV304 cell growth was suppressed by 76.51%, 71.09% and 61.49% after 48 h treatment with As2O3 at 0.5, 2.5 and 5 μmol/L, respectively. Early apoptosis in the As2O3- treated mice was 2.88-5.1 times higher than that in the controls, and late apoptosis was 1.17-1.67 times higher than that in the controls.CONCLUSION： Our results showed that As2O3 delays tumor growth, inhibits MVD, down-regulates Flt-1 and KDR expression, and disturbs the stimulating effect of VEGF165 on the growth of SGC7901 cells. These results suggest that As2O3 might delay growth of gastric tumors through inhibiting the paracrine and autocrine pathways of VEGF/VEGFRs.展开更多
Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate,to detect the level fluctuation of reactive oxygen species (ROS) during the cel...Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate,to detect the level fluctuation of reactive oxygen species (ROS) during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry,with TdT mediated dUTP nick end labeling (Tunel) and propidium iodide (PI),indicated that As2O3 (2 μM) could induce apoptosis in NB4 cells prevailingly from G2/M phase,and this efficacy was enhanced upon co-administration of 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) (2.5 μM) which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determin the selective induction of G2/M apoptosis and the cells' susceptibility to apoptosis by As2O3.展开更多
文摘Objective:To explore the cardioprotective effect of hesperidin against arsenic trioxide-induced cardiac toxicity in rats.Methods:Cardiac toxicity was induced by oral administration of 4 mg/kg arsenic trioxide for 30 days.Hematological,biochemical,electrocardiography,echocardiography,and histopathological examinations were performed.Results:Hesperidin decreased the neutrophil-to-lymphocyte ratio,calcium,creatine kinase-myoglobin binding,lactate dehydrogenase,IL-6,and lipid peroxidation,as well as increased sodium and potassium concentration and superoxide dismutase and catalase activity in arsenic trioxide-intoxicated rats.Moreover,it reduced peak systolic velocity and end-diastolic velocity while increasing heart rate.Arsenic trioxide-induced histopathological damage to cardiac tissue was prominently alleviated by hesperidin treatment.Conclusions:Hesperidin attenuates arsenic trioxide-induced cardiac toxicity in rats.Therefore,it can be further explored as a cardioprotective agent.
基金supported by a grant from theNational Natural Science Foundation of China(No.30572114).
文摘OBJECTIVE To study the anti-tumor effect of arsenic trioxide on the HepG2 human hepatocellular carcinoma cell line, and to explore its mechanism of action. METHODS The MTT assay was used to determine the inhibitory effect of As2O3 on HepG2 cells at various As2O3 concentrations. The expression of p-JNK, caspase-3 and PARP was detected by Western blots. RESULTS As2O3 markedly inhibited the growth of the HepG2 cells and induced apoptosis. The results of Western blot analysis showed that the As2O3-induced apoptosis was accompanied by caspase-3 and PARP activation. p-JNK was detected at 10 min following As2O3 treatment, and preceded to peak at 20 min, and decreased by 30 min. The total protein content did not obviously change. The activation of JNK occurred prior to cell apoptosis. SP600125, a JNK inhibitor, suppressed the As2O3-induced activation of caspase-3 and PARP cleavage. CONCLUSION As2O3 inhibits the proliferation of human HepG2 hepatocellular carcinoma cells by inducing apoptosis in vitro. As2O3-induced apoptosis is accessed through the caspase-3 pathway. The JNK signal-transduction pathway and caspase-3 are involved upstream in the As2O3 induced HepG2 apoptotic response.
基金Heilongjiang Natural Science Foundation (G98L19-1)guided by Ministry of Health,China.98-2-269
文摘AIM To study the effect of a varyingconcentrations of arsenic trioxide on humanhepatoma cell line BEL-?402 cultured in vitro andits mechanism of action.METHODS The BEL-7402 cells were treatedwith arsenic trioxide（at the concentrations of0.5,1,2 μmol/L,respectively）for 4 successivedays.The cell growth and proliferation wereobserved by cell counting and cell-growth curve.Morphologic changes were studied withelectronmicroscopy.Flow cytometry was usedto assay celI-DNA distribution and the proteinexpression of Bcl-2 and Bax detected byimmunocytochemical method.RESULTS The cell growth was significantlyinhibited by varying concentrations of arsenictrioxide as revealed by cell counting and cell-growth curve,which was dose- and time-dependent.Arsenic trioxide treatment at 0.5,1and 2 μmol/L resulted in a sub-G1 cell peak,theapoptosis rate of the control group was 9.31%and that of 0.5 μmol/L arsenic trioxide 15.53%,no significant difference was seen between thetwo.The apoptosis rates of 1,2 μmol/L arsenictrioxide were 19.10% and 21.87% respectively,which were much higher（both P【0.05）.Decrease of G<sub>0</sub>/G<sub>1</sub> phase cells and increase of Sphase cells were observed by flow cytometry,suggesting the inhibition effect of 0.5,1,2 μmol/L arsenic trioxide on BEL-7402 cell lay in the G<sub>0</sub>/G<sub>1</sub> phase.Morphologic changes such asintact cell membrane,nucleic condensation,apoptotic body formation were seen undertransmission electronmicrescopy,whereas the0.5 mol/L arsenic trioxide-treated BEL-7402cells showed decrease of nucleocytoplasmicratio,round nucleus,well-differentiatedorganelles in the cytoplasm.The processes andmicrovilli on the cell surface of the experimentalgroups under scanning electron microscopy weresignificantly decreased.High expressions ofBcl-2 and Bax were detected in 1 and 2 μmol/Larsenic trioxide-treated cells,these were 46%,87.33% and 83.08%,95.83% respectively,among which that of Bax was more significant.Arsenic trioxide treatment at 0.5 μmol/Lresulted in a higher expression level of Bcl-2 andlower expression level of Bax,which were8.81% and 3.83% respectively,as comparedwith that of the control group（15.33%）（P<sub>1</sub>【0.01,P<sub>2</sub>【0.01）.CONCLUSION Arsenic trioxide not onlyinhibited proliferation but also induced apoptosisof human hepatoma cell line BEL-7402.Theinduced-apoptosis effect of 1,2 μmol/L arsenictrioxide was related to the expression level ofBcl-2 and Bax.
文摘AIM： To study the anti-hepatoma efficiency of arsenic trioxide （As2O3） in the treatment of experimental rat hepatocellular carcinoma （HCC） induced by 2-acetamidofluorene （2-FAA） and to elucidate the possible mechanisms. METHODS： SD rats （2 mo old） had been fed with 2-FAA for 8 wk to induce HCC, and then they were treated with As2O3 or matrine. On d 29, the rats were killed and the liver was weighed and liver tumors were counted. The histological changes of liver tissue were observed under microscope, and the cellular dynamic parameters were studied by flow cytometry. Immunohistochemistry （two-step method） was used to observe the expression of vascular endothelial growth factor （VEGF） and micro-vessel density （MVD） on consecutive sections. The pathological parameters were also analyzed, the levels of serum aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, total bilirubin （TBi）, and direct bilirubin （DBi）. RESULTS： The number of liver tumors decreased significantly in groups treated with As2O3, especially in medium-dose （1 mg/kg） group （t = 2.80, P〈0.01）. As2O3 caused HCC cell death via apoptosis; necrosis was seen and apoptosis was common when the dose was 1 mg/kg. Proliferation index decreased sharply in medium-dose （1 mg/kg） group （7.87±4.11 vs24.46±6.49, t= 2087, P〈0.01）, but not in 0.2 mg/kg group. However, S-phase fraction decreased dramatically in both groups, it reached the bottom level only when the dose was i mg/kg compared with control （0.40±0.13 vs3.01±0.51, t= 2.97, P〈0.01）, and it was obviously accompanied with accumulation of cells in G0/G1 （G0/G1 restriction）. The expressions of VEGF and MVD in medium-dose （1 mg/kg） group were significantly lower than normal saline group （0.63±0.74 vs2.44±0.88, P〈0.05; 15.75±3.99 vs47.44±13.41, t= 2.80, P〈0.01）. Compared with normal saline group, mediumand low-dose groups As203 and matrine lowered the levels of ALT in serum （61.46±9.46, 63.75±20.40, 61.18±13.00 vs 108.98±29.86, t= 2.14, P〈0.05）, but had no effect onthe level of serum AST, TBi, and DBi. CONCLUSION： As203 had inhibitory effect on growth of experimental HCC in rats induced by 2-FAA, but had no obvious effect on normal hepatic cells. The mechanisms may involve decrease of cell division, accumulation of cells in G0/G1 phase, apoptosis of tumor cells, and inhibitory effect on angiogenesis through blocking VEGF.
基金Supported by the Key Scientific and Technological Projects of Heilongjiang Province during the 10~(th) Five-Year Plan Period, No. GB05C401-10
文摘AIM： To investigate the effect of all-trans-retinoic acid （ATRA） on arsenic trioxide （As2O3）-induced apoptosis of human hepatoma, breast cancer, and lung cancer cells in an attempt to find a better combination therapy for solid tumors. METHODS： Human hepatoma cell lines HepG2, Hep3B, human breast cancer cell line MCF-7, and human lung adenocarcinoma cell line AGZY-83-a were treated with As203 together with ATRA. Cell survival fraction was determined by MTT assay, cell viability and apoptosis were measured by annexin V-fluorescein isothiocyanate （FITC） and PI staining, and intracellular glutathione （GSH） and glutathione-S-transferase （GST） activities were determined using commercial kits. RESULTS： Cytotoxicity of ATRA was low. ATRA （0.1, 1, and 10 μmol/L） could synergistically potentiate As2O3 to exert a dose-dependent inhibition of growth and to induce apoptosis in each of the cell lines. HepG2 and Hep3B with low intracellular GSH or GST activities were remarkably sensitive to As2O3 or As2O3＋ATRA, while AGZY-83-a with higher GSH or GST activities was less sensitive to As2O3 or As2O3＋ATRA. Treatment with 2 μmol/L As2O3 for 72 h significantly decreased intracellular GSH and GST levels in each of the cell lines, and 1 μmol/L ATRA alone reduced minimal intracellular GSH and GST levels. ATRA potentiated the effect of As2O3 on intracellular GSH levels, but intracellular GST levels were not significantly affected by the combination of As2O3 and ATRA for 72 h as compared to As2O3 alone.CONCLUSION： ATRA can strongly potentiate As2O3- induced growth-inhibition and apoptosis in each of the cell lines, and two drugs can produce a significant synergic effect. The sensitivity to As2O3 or As2O3＋ATRA is inversely proportional to intracellular GSH or GST levels in each of the cell lines. The GSH redox system may be the possible mechanism by which ATRA synergistically potentiates As203 to exert a dose-dependent inhibition of growth and to induce apoptosis. 2005 The WJG Press and Elsevier Inc. All rights reserved.
文摘BACKGROUND:It has been pointed out that only low-dose arsenic trioxide(ATO)presents therapeutic benefits outweighing the toxic side effects.Low-dose ATO can effectively alleviate acute promyelocytic leukemia(APL). However,it is quite challenging in treating solid tumors. The purpose of this study was to investigate the effect of ATO at low concentrations on the metastatic potential of mouse hepatoma H22 cells and the anti-metastatic mechanism of ATO. METHODS:The metastatic potential of H22 cells was evaluated by adhesion,migration and invasion assays after exposure to a low dose of ATO in vitro.The mouse lung metastatic model induced by injection of H22 cells via the tail vein was adopted for the evaluation of metastatic potential. Different proteins in the lysate of H22 cells exposed to ATO at different concentrations were investigated by surface- enhanced laser desorption and ionization time-of-flight mass spectrometry(SELDI-TOF-MS).Finally,Western blotting analyses were made to detect the expression pattern of MMP-2 and nm23-M1 proteins. RESULTS:Significant cell death started at ATO concentrations above 2μmol/L.The growth and adhesion potential of H22 cells was inhibited in a time-and dose- dependent manner,and the migration and invasion potential of H22 cells was inhibited in a dose-dependent manner while ATO concentration was below 2μmol/L. Mice injected with ATO at a dose of 0.5 mg/kg had fewer lung metastases.However,mice injected with ATO at a dose of 2 mg/kg or 4 mg/kg had a high mortality rate and more liver injuries.A total of 15 different protein peaks were identified between the lysate of H22 cells treated with ATO and controls.Two proteins that peaked atm/z 5302 and 17207 coincided with MMP-2(fragment) and nm23-M1,respectively.Western blotting analyses demonstrated that MMP-2 and MMP-2 fragments were down-regulated and nm23-M1 was up-regulated in H22 cells treated with 2μmol/L ATO for 48 hours. CONCLUSIONS:ATO at a low dose inhibits the metastatic potential of mouse hepatoma H22 cells in vitro and in vivo, and involves down-regulation of MMP-2 and up-regulation of nm23-M1.
基金funding support from the National Natural Science Foundation of China (No. 81873014)。
文摘Objective To reduce the toxicity and side effects of arsenic trioxide(ATO)and provide a new approach for the treatment of primary liver cancer,a folic acid-modified calcium arsenite liposomal“target-controlled”drug delivery system(FA-LP-CaAs)was fabricated using the reverse microemulsion method.Methods A Malvern particle size analyzer and a transmission electron microscope were employed to determine the particle size,distribution,zeta potential and morphology of FA-LP-CaAs.Further,inductively coupled plasma emission spectrometry was employed to determine the drug loading capacity,entrapment efficiency,and in vitro release behavior of FA-LP-CaAs.To determine its toxicity in human hepatoma cells(HepG2)and human normal hepatocytes(LO2)and its effect on HepG2 cell cycle and apoptosis,the MTT method was used.Laser confocal and flow cytometry were also employed to determine the uptake of FA-LP-CaAs by cells.After establishing a mouse liver cancer model,the in vivo distribution of the drug included in the formulation was investigated using in vivo fluorescence.To evaluate the liver cancer targeting and anti-tumor effects of FALP-CaAs in vivo,the distribution of ATO in tissues and changes in tumor volume and body weight after liposomal administration were investigated using hematoxylin-eosin(HE)-stained tumor sections.Results The particle size,zeta potential and PDI of FA-LP-CaAs were(122.67±2.18)nm,(12.81±0.75)mV and 0.22±0.01,respectively,while its drug loading capacity was 18.49%±1.14%.In vitro experimental results revealed that FA-LP-CaAs had a strong killing effect on HepG2 cells.Further,the cell uptake capacity of this formulation was found to improve.Based on in vivo assessments,FA-LP-CaAs could significantly increase the distribution of ATO in tumor sites and inhibit tumor growth.Conclusions Herein,an FA-LP-CaAs formulation was successfully fabricated.This liposomal drug delivery system had a round appearance,uniform particle size,good polydispersity coefficient,evident“core-shell”structure,high drug loading capacity and pH response,tumor targeted drug delivery and sustained drug release.These findings support further research and the application of ATO as an anti-liver cancer prodrug and provide a new method for the treatment of liver cancer.
基金This work was supported by the grantform the Natural Foundation of Guang-dong Province (No. 021195) and Guang-zhou City (No. 2001-Z-037-01).
文摘OBJECTIVE To explore the inhibitory effect of arsenic trioxide on growth and telomerase activity of BEL-7402 and SMMC-7721 hepatocarcinoma cells, and to measure their GSH level. METHODS Cell culture and trypan blue exclusion were used to examine the inhibitory effect of arsenic trioxide on BEL-7402 and SMMC-7721 hepatocarcinoma lines. A GSH kit and telomerase kit were used to mearsure the GSH content in cells and telomerase activity. RESULTS The growth of BEL-7402 cells was significantly inhibited at a level of 0.50μmol/L of arsenic trioxide by 24 h. The inhibitory effect increased with time and concetration of arsenic trioxide. The telomerase activity of BEL-7402 cells was also significantly inhibited at a level of 0.50μmol/L of arsenic trioxide by 24 h, after which the inhibitory effect increased with time. On the other hand, at 24 h of incubation a level of 2.00 μmol/L of arsenic trioxide was required to significantly inhibit growth of SMMC-7721 cells, and only after 48 h with 2.00μmol/L of arsenic trioxide did telomerase activity significantly decline. The GSH content of the BEL-7402 and SMMC-7721 cells was 18.7±1.4 and 50.8±5.2 nmol/mg protein respectively, a significant difference. CONCLUSION Different concentrations of arsenic trioxide are required to inhibit growth and telomerase activity of SMMC-7721 and BEL-7402 cells. Perhaps BEL-7402 cells are more sensitive to arsenic trioxide because of their low level of GSH content, which results in a low capacity for oxidation-reduction and poorer detoxification mechanisms in BEL-7402 cells.
基金supported by a grant from the Army Logistics Research Plan of China (AEP14C001)National Natural Science Foundation of China (81472478)Public Science and Technology Research Funds Projects of Quality Inspection (201510024)
文摘Objective: The study aimed to investigate the correlation between concentration of inhaled arsenic trioxide and dynamic changes in hematotoxicity in rats. Method: Wistar rats were randomly divided into four study groups that were treated with saline(control) or arsenic trioxide at a low(0.1 mg/mL), medium(1 mg/mL), or high(10 mg/mL) dose by intratracheal instillation. Blood samples were collected for analysis at 6, 12, 24, 48, and 72 h after exposure. Results: Compared with the control group, the red blood cell distribution width decreased significantly in each exposure group and then gradually recovered. The red blood cell count only increased in the high-dose group, and the hemoglobin levels did not differ significantly between the groups. Significant decreases in the platelet count and mean platelet volume were observed in each arsenic-exposed group. White blood cell countsalso decreased significantly and then gradually recovered; this was associated with an increased percentage of lymphocytes and a decreased percentage of neutrophils. The blood cell parameter changes were related to the arsenic trioxide concentration and the observation time. Conclusion: Intratracheal instillation of arsenic trioxide affected hematopoietic differentiation in rats, leading to blood cell changes that were related to observation time and concentration.
基金supported by a grant from the Science and Technology Committee of Sichuan Province (No2008JY0029-1)
文摘Objective： To explore the effects of arsenic trioxide （ATO） on the apoptosis of glucocorticoid （GC）-resistant T-acute lymphoblastic leukemia （ALL） CEM-C1 cells and its possible mechanisms. Methods： Different concentrations of ATO （0.25 μmol/L-5 μmol/L） were used to induce the apoptosis of CEM-C1 cells. The inhibition rate of cell proliferation and apoptosis were detected by MTT test, Annexin V-FITC/PI flow cytometry and optical microscopy, respectively. RT-PCR was applied to semi-quantitatively analyze the mRNA expression of pro-apoptotic proteins （Bad and PDCD4） and anti-apoptotic proteins （XIAP and MCL-1） induced by different concentrations of ATO at different time points. Results： ATO could inhibit proliferation and induce apoptosis of CEM-C1 cells at a concentration and time dependent manner. Low-dose ATO mildly inhibited the proliferation of CEM-C1 cells while higher concentrations （1 μmol/L and 5 μmol/L） had strong anti-tumor effect with the inhibiting rates of 40.07±7.98% and 88.67±2.88%, respectively. Annexin V-FITC/PI flow cytometry showed that the apoptotic rates of CEM-C1 ceils were significantly increased after 48 hours treatment of different concentrations of ATO. RT-PCR demonstrated up-regulated mRNA expression of pro-apoptotic protein Bad and PDCD4 but down-regulated mRNA expression of anti-apoptotic protein XIAP when CEM-C1 cells were treated with different concentrations of ATO at different time points. The MCL-1 mRNA expression was down-regulated only after the treatment of 5 μmol/L ATO. Conclusion： ATO can inhibit cell proliferation and induce cell apoptosis in GC-resistant CEM-C1 cells. The molecular mechanisms might involve the increased mRNA expression of pro-apoptotic protein Bad and PDCD-4, and rapid down-regulation of XIAP mRNA expression.
基金National Natural Science Foundation of China(No.30170368)
文摘Aim This study was to evaluate the effect of arsenic trioxide （As2O3） on the transgenic TNF-α promoter activity in cultured vascular smooth muscle cells （VSMCs） and THP-1 monocytes. Methods Human TNF-α promoter was constructed by reporter gene system and was transiently transfected into VSMCs and THP-1 in vitro. The promoter activity was tested by luciferase activity with or without LPS and Ang Ⅱ stimulation, before and after different dosage of As2O3 treatment. Results 1. TNF-α promoter effectively expressed in VSMCs and THP-1 compared with CMV promoter （58.3% and 80.9%, respectively）. Both LPS and Ang Ⅱ significantly up-regulated TNF-α promoter activity （P〈0.05）. 2. As2O3 significantly inhibited, both intact and LPS/Ang Ⅱ stimulated promoter activity, in a dose dependent manner （P〈0.05）, and in both cell type. Conclusion These results manifested that, the inhibition effect of As2O3 on the activity of human TNF-α promoter indicated its potential inhibition on pro-inflammatory cytokine genes expression at transcriptional level and its potential anti-inflammatory property in the cardiovascular system.
文摘Roles of Marigold extracts (ME) on arsenic trioxide (ATO)-induced oxidative damage to pancreatic β-cells need to be further elucidated. In this study, NIT-1 cells were treated with different concentrations of and/or ATO, following by the cell viability was detected by CCK8 assay. Then, intracellular reactive oxygen species (ROS) levels, lipid peroxide (MDA) contents and superoxide dismutase (SOD) activity were measured with a fluorescence probe method and colorimetric assay, respectively. The apoptosis rate and morphology was detected and observed with hoechst 33,258 staining assay. The mRNA levels and protein expressions of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were measured by real-time fluorescence quantitative polymerase chain reaction and protein immunoblotting assay, respectively. Our results indicated that Co-treatment with ME and ATO exacerbated the cell viability decreasing reduced by ATO, while the addition of ME after ATO treatment effectively promote the recovery of ATO reduced survival rates. The ATO group increased apoptosis (P P β-cells by modulating the activation of the Nrf2 signaling pathway.
基金the Natural Science Foundation of Committee of Science and Technology of Shanghai Municipality(№964119035)
文摘AIM To study the effects of arsenic trioxide andHCPT on different degrees of differentiated gastriccancer cells（SGC-7901,MKN-45,MKN-28）withrespect to both cytotoxicity and induction ofapoptosis in vitro.METHODS The cytotoxicity of As<sub>2</sub>O<sub>3</sub> and HCPTon gastric cancer cells was determined by MTTassay.Morphologic changes of apoptosis ofgastric cancer cells were observed by lightmicroscopy and transmission electron microscopy.Apoptosis and cell cycle changes of gastric cancercells induced by HCPT and As<sub>2</sub>O<sub>3</sub> were investigatedby TUNEL method and flow cytometry.RESULTS As<sub>2</sub>O<sub>3</sub> and HCPT had remarkablecytotoxic effects on different degrees ofdifferentiated gastric cancer cells.The IC<sub>50</sub>ofAs<sub>2</sub>O<sub>3</sub> on well differentiated gastric cancer cellMKN-28,moderately differentiated gastric cancercell SGC-7901,and poorly differentiated gastriccancer cell MKN-28 were 8.91 μmol/L,10.57μmol/L,and 11.65 μmol/L,respectively.The IC<sub>50</sub>of HCPT on MKN-28,SGC-7901,and MKN-45 were9.35 mg/L,10.21 mg/L,and 12.63 mg/Lrespectively after 48 h treatment.After 12 h ofexposure to both drugs,gastric cancer cellsexhibited morphologic features of apoptosis,including cell shrinkage,nuclear condensation, and formation of apoptotic bodies.A typicalsubdiploid peak before G<sub>0</sub>/G<sub>1</sub> phase was observedby flow cytometry.The apoptotic rates of SGC-7901,MKN-45,and MKN-28 were 13.84%,22.52%,and 9.68%,respectively after 48 hexposure to 10 μmol/L As<sub>2</sub>O<sub>3</sub>.The apoptotic ratesof SGC-7901,MKN-45,and MKN-28 were 21.88%,12.35%,and 30.26%,respectively after 48 hexposure to 10 mg/L HCPT.The apoptotic indicewere 7%-15% as assessed by TUNEL method.The effect of As<sub>2</sub>O<sub>3</sub> on SGC-7901 showedremarkable cell cycle specificity,which inducedcell death in G<sub>1</sub> phase,and blocked G<sub>2</sub>/M phase.HCPT also showed a remarkable cell cyclespecificity,by inducing cell death and apoptosis inG<sub>1</sub> phase and arrest of proliferation at S phase.CONCLUSION As<sub>2</sub>O<sub>3</sub> and HCPT exhibitsignificant cytotoxicity on gastric cancer cells byinduction of apoptosis.As<sub>2</sub>O<sub>3</sub> and HCPT mighthave a promising prospect in the treatment ofgastric cancer,which needs to be further studied.
文摘Since arsenic trioxide was first approved as the front line therapy for acute promyelocytic leukemia 25 years ago,its anti-cancer properties for various malignancies have been under intense investigation.However,the clinical successes of arsenic trioxide in treating hematological cancers have not been translated to solid cancers.This is due to arsenic＇s rapid clearance by the body＇s immune system before reaching the tumor site.Several attempts have henceforth been made to increase its bioavailability toward solid cancers without increasing its dosage albeit without much success.This review summarizes the past and current utilization of arsenic trioxide in the medical field with primary focus on the implementation of nanotechnology for arsenic trioxide delivery to solid cancer cells.Different approaches that have been employed to increase arsenic＇s efficacy,specificity and bioavailability to solid cancer cells were evaluated and compared.The potential of combining different approaches or tailoring delivery vehicles to target specific types of solid cancers according to individual cancer characteristics and arsenic chemistry is proposed and discussed.
基金Supported by the Zhejiang Province Science and Technology Fund for excellent returnee 2001-275
文摘AIM: To investigate the effect of arsenic trioxide on human gastric cancer cell line MKN45 with respect to both cytotoxicity and induction of apoptosis in vitro. METHODS: MKN45 cells were treated with arsenic trioxide (As2O3) at the concentration of 1, 5, and 10 μmol/L, respectively, for three successive days. Cell growth and proliferation were observed by cell counting and trypan blue exclusion. Cytotoxicity of As2O3 was determined by MTT assay. Morphologic changes were studied with light microscopy. Flow cytometry was used to assay cell DNA distribution and apoptotic cells were confirmed with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and DNA electrophoresis. RESULTS: The growth of MKN45 cells was significantly inhibited by As2O3 which was confirmed by colony-forming assay. After 7 d of culture with various concentrations of As2O3, colony-forming capacity of MKN45 cells decreased with As2O3 increment in comparison with that of control group. The inhibitory rate of colony-formation was 38.5%, 99.1%, and 99.5% when the concentration of As2O3 was 1, 5, and 10 μmol/L in culture medium, respectively. The cell number of a single colony in drug treatment groups was less than that of control group. The cell-killing rate of As2O3 to MKN45 cells was both dose- and time-dependent with an IC50 of (11.05±0.25) μmol/L After incubation in 10 μmol/L As2O3 for 24 h, the cell-killing rate was 27.1%, and it was close to 50% after 48 h. The results showed that As2O3 induced time- and dose-dependent apoptosis in MKN45 cells, blocked at G2/M phase. The apoptotic peak (sub-G1 phase) appeared and cell apoptotic rate in MKN45 cells was 18.3-32.5% after treatment by 10 umol/L As2O3 for 48 h. The percentage of G2/M cell of the experimental groups was 2.0-5.0 times than that of the control group. Gel electrophoresis of DNA from cells treated with each concentration of As2O3 for 48 h revealed a 'ladder' pattern, indicating preferential DNA degradation at the internucleosomal, linker DNA sections. TUNEL also demonstrated strand breaks in DNA of MKN45 cells treated with As2O3, while control cells showed negative labeling. CONCLUSION: As2O3 can induce apoptosis of human gastric carcinoma cells MKN45, which is the basis of its effectiveness. It shows great potential in the treatment of gastric carcinoma.
基金Supported by The Heilongjiang Provincial Natural Science Foundation of China,No.D2006-51
文摘AIM: To investigate the changes in apoptosis in gastrointestinal cancer cells from patients with gastrointestinal cancers treated with arsenic trioxide (As<sub>2</sub>O<sub>3</sub>); and to study the possible molecular mechanisms of such changes by detecting the expression levels of p53 and Bcl-2.
基金Supported by the Science Fund of the Second Affiliated Hospital of Medical College, No. 2003-YL-35
文摘AIM: To investigate the inhibitory effect of As2O3 on angiogenesis of tumor and expression of vascular endothelial growth factor (VEGF) in tumor cells in vivo and in vitro. METHODS: The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were randomly divided into three groups. As2O3 was injected into the arsenic-treated groups (2.5 mg/kg and 5 mg/kg) and the same volume of saline solution was injected into the control group. Microvessel density (MVD) and expression of VEGF were detected with immunofluorescence laser confocal technology. Further expression of VEGF protein and VEGF mRNA was measured with Western bloting and fluorescence quantitative RT- PCR in SGC-7901 cells treated with As2O3. RESULTS: In nude mice, after treatment with 5 mg/kg and 2.5 mg/kg As2O3 respectively, about 50% and 30% tumor growth inhibition were observed correspondingly (P < 0.05, P < 0.05). Decrease in MVD appeared in As2O3-treated tumors compared with control group (P < 0.001, P < 0.001). MVD in tumors was significantly lower in 5 mg/kg group than in 2.5 mg/kg group (P < 0.01). The fluorescence intensity levels of VEGF in tumor cells were significantly lowered in the arsenic-treated groups (P < 0.01, P < 0.01). The fluorescence intensity level of VEGF in 5 mg/kg group was lower than that in 2.5 mg/ kg group (P < 0.01). In vitro, the expression of VEGF protein decreased in dose- and time-dependent manner after the treatment with As2O3, but in VEGF mRNA no significant difference was found between the control group and the treated groups. CONCLUSION: As2O3 can inhibit solid tumor growth by inhibiting the formation of new blood vessels. One of the mechanisms is that As2O3 can inhibit VEGF protein expression.
文摘INTRODUCTION Cell apoptosis,which involves the biologic regulation of the numbers and vital activity of cells,is an important metaboloc process in both normal cells and tumor cells.
基金Supported by The Science Fund of the Second Affiliated Hospital of the Medical College,No.2003-YL-35
文摘AIM： To investigate the effect of arsenic trioxide （As2O3） on expression of vascular endothelial growth factor receptor-1 （VEGFR-1, Flt-1） and VEGFR-2 （KDR） in human gastric tumor cells and proliferation of vascular endothelial cells.METHODS： The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were treated with As2O3. Microvessel density （MVD） and expression of Flt-1 and KDR were detected by immunofluorescence laser confocal microscopy. SGC-7901 cells were treated respectively by exogenous recombinant human VEGF165 or VEGF165 ＋ As2O3. Cell viability was measured by MTT assay. Cell viability of ECV304 cells was measured by MTT assay, and cell cycle and apoptosis were analyzed using flow cytometry.RESULTS： The tumor growth inhibition was 30.33% and 50.85%, respectively, in mice treated with As2O3 2.5 and 5 mg/kg. MVD was significantly lower in arsenic-treated mice than in the control group. The fluorescence intensity levels of Flt-1 and KDR were significantly less in the arsenic-treated mice than in the control group. VEGF165 may accelerate growth of SGC7901 cells, but As2O3 may disturb the stimulating effect of VEGF165. ECV304 cell growth was suppressed by 76.51%, 71.09% and 61.49% after 48 h treatment with As2O3 at 0.5, 2.5 and 5 μmol/L, respectively. Early apoptosis in the As2O3- treated mice was 2.88-5.1 times higher than that in the controls, and late apoptosis was 1.17-1.67 times higher than that in the controls.CONCLUSION： Our results showed that As2O3 delays tumor growth, inhibits MVD, down-regulates Flt-1 and KDR expression, and disturbs the stimulating effect of VEGF165 on the growth of SGC7901 cells. These results suggest that As2O3 might delay growth of gastric tumors through inhibiting the paracrine and autocrine pathways of VEGF/VEGFRs.
基金supported by research grants from National Natural Science Foundation of China(No.30170475)
文摘Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate,to detect the level fluctuation of reactive oxygen species (ROS) during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry,with TdT mediated dUTP nick end labeling (Tunel) and propidium iodide (PI),indicated that As2O3 (2 μM) could induce apoptosis in NB4 cells prevailingly from G2/M phase,and this efficacy was enhanced upon co-administration of 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) (2.5 μM) which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determin the selective induction of G2/M apoptosis and the cells' susceptibility to apoptosis by As2O3.
