[Objective] This study aimed to test the sensitivity of Eperythrozoon from mink to various drugs in vitro. [Method] The red blood cells isolated from Eperythro- zoon positive mink was cultured in complete medium （70%...[Objective] This study aimed to test the sensitivity of Eperythrozoon from mink to various drugs in vitro. [Method] The red blood cells isolated from Eperythro- zoon positive mink was cultured in complete medium （70% RPMI-1640 medium and 30% calf serum）, supplemented with bernier, oxytetracycline, trichlorfon, tylosin, imi- docarb, florfenicol, Fuhongjuesha or primaquine phosphate at the working concentra- tions of 12, 24, 36, 48 and 96 μL/ml, incubated at 37.3 ℃, 5% CO2. [Result] Fuhongjuesha was most efficient for killing Eperythrozoon, followed by trichiorfon and primaquine phosphate, but trichlorfon is toxic. Bernier, imidocarb and florfenicol are efficient. [Conclusion] The study provides a scientific reference for clinical treatment of eperythrozoonosis.展开更多
Objective: To study the expression of NFκB family protein in breast cancer cell lines and the relationship between NFκB family protein and the drug resistance.Methods: Expression of P65, IκB-α in 14 breast cancer ...Objective: To study the expression of NFκB family protein in breast cancer cell lines and the relationship between NFκB family protein and the drug resistance.Methods: Expression of P65, IκB-α in 14 breast cancer cell lines and P50 in 11 breast cancer cell lines was detected by Western blot. The sensitivity of the cells to ADM was determined by MTT.Results: 1κB-α located mainly in the cytoplasm. P65 and P50 were in both of cytoplasm and nucleus. The expression level of P50 was higher than that of P65, especially in nucleus. MTT assay showed that IC50 was three-fold higher in the cell lines which expressed P50 in nucleus than those P50 negative in nucleus, but no difference was found in the expression of P65.Conclusion: The expression of P50 in nucleus may predict the chemotherapy resistance in breast cancer, so it can be used as an indicator to predict the prognosis. The expression of P50 is more often in breast cancer, and it may play a more important role in the chemotherapy resistance than other NFκB family members. Key words breast cancer - NFκB - chemotherapy resistance展开更多
AIM： To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy. METHODS： Three gastric cancer cell lines with distinct levels of differen...AIM： To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy. METHODS： Three gastric cancer cell lines with distinct levels of differentiation were treated with various doses of rAd-p53 alone, oxaliplatin （OXA） alone, or a combination of both. Cell growth was assessed with an 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-diphenytetrazoli- umromide assay and the expression levels of p53, Bax and Bcl-2 were determined by immunohistochemistry. The presence of apoptosis and the expression of caspase-3 were determined using flow cytometry. RESULTS： Treatment with rAd-p53 or OXA alone inhibited gastric cancer cell growth in a time- and dose- dependent manner; moreover, significant synergistic effects were observed when these treatments were combined. Immunohistochemical analysis demonstrated that treatment with rAd-p53 alone, OXA alone or combined treatment led to decreased Bcl-2 expression and increased Bax expression in gastric cancer cells.Furthermore, flow cytometry showed that rAd-p53 alone, OXA alone or combination treatment induced apoptosis of gastric cancer cells, which was accompa- nied by increased expression of caspase-3. CONCLUSION： rAd-p53 enhances the sensitivity of gastric cancer cells to chemotherapy by promoting apoptosis. Thus, our results suggest that p53 gene therapy combined with chemotherapy represents a novel avenue for gastric cancer treatment.展开更多
Objective:To address how genistein sensitizes the chemotherapy-resistant ovarian carcinoma cells and promotes apoptosis in the respect of cell cycle and the regulation of survivin expression in the process.Methods:Ova...Objective:To address how genistein sensitizes the chemotherapy-resistant ovarian carcinoma cells and promotes apoptosis in the respect of cell cycle and the regulation of survivin expression in the process.Methods:Ovarian SKOV-3 carcinoma cell line was treated with genistein or cisplatin either alone or in combination.Cell viability was showed by MTT method.Cell cycle and apoptosis were detected by flow cytometry.Survivin mRNA and protein were revealed by RT-PCR and immunocytochemistry,respectively.Results:Genistein could reduce the cell viability in a dose-dependent manner,while cisplatin did so at a much higher level.In contrast,if the two agents were treated in combination,half growth inhibition(IC50) value for cisplatin was reduced remarkably and the effect was synergistic as analyzed by isobologram.In particular,the reduced cell viability was exhibited by a switch in cell cycle progression,as the cells were arrested in G2/M phase and the G0/G1 phase-fraction was significantly decreased.The reduced cell viability appeared to involve apoptosis,based on our results from flow cytometry and Hoechst 33258 staining.In the meanwhile,genistein performed the inhibitory effect on cisplatin-induced survivin expression.Conclusion:Genistein can sensitize ovarian carcinoma cells to cisplatin therapy with the inhibition of survivin expression as the potential mechanism.展开更多
Objective: To investigate the association of the transportation characteristics of nolatrexed in tumor cells with its drug sensitivity. Methods: The sensitivity of 3 tumor cell lines, C6, SRS82 and LoVo, to nolatrexed...Objective: To investigate the association of the transportation characteristics of nolatrexed in tumor cells with its drug sensitivity. Methods: The sensitivity of 3 tumor cell lines, C6, SRS82 and LoVo, to nolatrexed were determined by growth inhibition study. After exposure to 20 μmol/L nolatrexed at different time intervals ranging from 0 to 30 min, or to nolatrexed at different concentrations ranging from 0 to 40 μmol/L for 10 min, the intracellular drug concentration was measured using high-performance liquid chro-matography. Results: C6 was the most sensitive cell line among the three, with sensitivity 6. 8-fold and 13. 8-fold those of SRS-82 and LoVo cells respectively. Transportation of nolatrexed in the 3 cell lines were qualitatively similar, which rapidly achieved steady-state within 5 min, and linear relationship between the intracellular and extracellular drug concentration was observed. The intracellular steady-state level achieved in C6 was significantly higher than those in the other two cell lines, the latter having comparable levels. Conclusion: Nolatrexed enters the cell very quickly and different transport capacities are involved in the generation of varied sensitivity to nolatrexed in tumor cells.展开更多
Objective:The aim of this study was to investigate the sensitivity of chemotherapeutic agents 5-FU,cisplatin(DDP) or TAXOL on colon cancer cell line SW480 with different methods,to find out the best examine time perio...Objective:The aim of this study was to investigate the sensitivity of chemotherapeutic agents 5-FU,cisplatin(DDP) or TAXOL on colon cancer cell line SW480 with different methods,to find out the best examine time period for further study of chemotherapeutic sensitivities.Methods:The SW480 cell was treated with 5-FU(200μg/mL),DDP(150μg/mL) or TAXOL(50μg/mL) respectively for 4h,8h or 12h.MTT assay was used to examine the cell survival rate,Annexin V/PI assay was used to analysis the apoptosis rate,Western Blot assay was applied to examine the expression of apoptotic protein.Results:(1) Results of MTT assay showed that the survival rate of SW480 cells at 4h,8h or 12h was:5-FU(200μg/mL)96.0%±8.2%,85.4%±7.8%,74.4% ±10.2%(P<0.05);DDP(150μg/mL) 99.0%±6.4%,88.7%±4.7%(P<0.05),46.9%±2.6%(P<0.01);TAXOL(50μg/mL) 51.5%±4.2%(P<0.01),31.9%±3.1%,17.6%±2.3%,or blank group 97.2%±5.8%,98.7%±7.2%,97.5%±7.5% respectively.(2) The apoptosis rate of cancer cells at 4h,8h or 12h was:control group:3.4%±0.2%,6.2%±0.4%,7.0%±0.5%;5-FU(200μg/mL) 4.0%±0.3%,4.8%±0.4%,7.7%±0.7%;DDP(150μg/mL) 8.5%±0.9%,18.6%±1.6%(P<0.05),67.0%±6.2%(P<0.01);or TAXOL(50μg/mL) 32.0%±5.2%(P<0.01),76.6%±8.5%,94.0% ±8.2%,respectively.(3) Western Blot assay showed that the expression of apoptosis associated protein.PARP,X-linked inhibitor of apoptasis(XIAP),Caspase-9 and Bcl-xL were changed.Conclusion:The sensitivity of chemotherapy could be assessed by MTT assay,Annexin V/PI assay and Western Blot.The best examine time of the three chemotherapuc agents was 5-FU(200μg/mL):>12h,DDP(150μg/mL):8-12h,or TAXOL(50μg/mL):<4h.展开更多
OBJECTIVE To explore the chemotherapeutic susceptibility of SP cells in human pulmonary adenocarcinoma cell line A549 and the possible mechanism of multidrug resistance.METHODS SP and non-SP （NSP） cells in the cell ...OBJECTIVE To explore the chemotherapeutic susceptibility of SP cells in human pulmonary adenocarcinoma cell line A549 and the possible mechanism of multidrug resistance.METHODS SP and non-SP （NSP） cells in the cell line A549 were isolated by fluorescence activated cell sorter. The susceptibility of SP and NSP cells to DDP, 5-FU, VP16, NVB and GEM was detected by a drug susceptibility test, and IC50s were calculated 24 h a er the chemotherapy; and then a er a 2-hour IC50 treatment with 5 chemotherapeutic drugs on the 2 subsets of NSP cells, the intracellular drug levels were determined and analyzed using high performance liquid chromatograph.RESULTS There was no statistical signifi cance in comparison of the di. erences in IC50s and in intracellular drug levels a er DDP treatment between the 2 subsets （P 〉 0.05）, （P 〉 0.05）. However,all IC50s of the other 4 drugs were significantly higher in the SP cells than in the NSP cells （P 〈 0.01）. A er the chemotherapy, the intracellular drug levels of the other 4 drugs were significantly lower in SP cells than in NSP cells （P 〈 0.01）.CONCLUSION Compared to NSP cells, SP cells in the cell line A549 have stronger resistance to the chemotherapeutics. The multidrug resistance of SP cells closely correlates with the function of SP cells discharging chemotherapeutic agents.展开更多
K-ras is a member of ras gene family which is involved in cell survival,proliferation and differentiation.When a mutation occurs in ras gene,the activation of Ras proteins may be prolonged to induce oncogenesis.Howeve...K-ras is a member of ras gene family which is involved in cell survival,proliferation and differentiation.When a mutation occurs in ras gene,the activation of Ras proteins may be prolonged to induce oncogenesis.However,the relationship between K-ras mutation and clinical outcomes in pancreatic cancer patients treated with chemotherapy agents is still under debate.In this study,we constructed five pAcGFP1-C3 plasmids for different types of K-ras gene(WT,G12V,G12R,G12D,and G13D)and stably transfected human pancreatic cancer Bxpc-3 cells with these genes.The wild type and mutant clones showed a comparable growth and expression of K-Ras-GFP fusion protein.The expression of some K-ras mutations resulted in a reduced sensitivity to gefitinib,5-FU,docetaxel and gemcitabine,while showed no effects on erlotinib or cisplatin.Moreover,compared with the wild type clone,K-Ras downstream signals(phospho-Akt and/or phospho-Erk)were increased in K-ras mutant clones.Interestingly,different types of K-ras mutation had non-identical K-Ras downstream signal activities and drug responses.Our results are the first to reveal the relationship between different K-ras mutation and drug sensitivities of these anti-cancer drugs in pancreatic cancer cells in vitro.展开更多
基金Supported by Fund of Hebei Science and Technology Department(10960408D01)~~
文摘[Objective] This study aimed to test the sensitivity of Eperythrozoon from mink to various drugs in vitro. [Method] The red blood cells isolated from Eperythro- zoon positive mink was cultured in complete medium （70% RPMI-1640 medium and 30% calf serum）, supplemented with bernier, oxytetracycline, trichlorfon, tylosin, imi- docarb, florfenicol, Fuhongjuesha or primaquine phosphate at the working concentra- tions of 12, 24, 36, 48 and 96 μL/ml, incubated at 37.3 ℃, 5% CO2. [Result] Fuhongjuesha was most efficient for killing Eperythrozoon, followed by trichiorfon and primaquine phosphate, but trichlorfon is toxic. Bernier, imidocarb and florfenicol are efficient. [Conclusion] The study provides a scientific reference for clinical treatment of eperythrozoonosis.
文摘Objective: To study the expression of NFκB family protein in breast cancer cell lines and the relationship between NFκB family protein and the drug resistance.Methods: Expression of P65, IκB-α in 14 breast cancer cell lines and P50 in 11 breast cancer cell lines was detected by Western blot. The sensitivity of the cells to ADM was determined by MTT.Results: 1κB-α located mainly in the cytoplasm. P65 and P50 were in both of cytoplasm and nucleus. The expression level of P50 was higher than that of P65, especially in nucleus. MTT assay showed that IC50 was three-fold higher in the cell lines which expressed P50 in nucleus than those P50 negative in nucleus, but no difference was found in the expression of P65.Conclusion: The expression of P50 in nucleus may predict the chemotherapy resistance in breast cancer, so it can be used as an indicator to predict the prognosis. The expression of P50 is more often in breast cancer, and it may play a more important role in the chemotherapy resistance than other NFκB family members. Key words breast cancer - NFκB - chemotherapy resistance
基金Supported by Xuzhou Science and Technology Development Fund,No. XM07C039
文摘AIM： To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy. METHODS： Three gastric cancer cell lines with distinct levels of differentiation were treated with various doses of rAd-p53 alone, oxaliplatin （OXA） alone, or a combination of both. Cell growth was assessed with an 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-diphenytetrazoli- umromide assay and the expression levels of p53, Bax and Bcl-2 were determined by immunohistochemistry. The presence of apoptosis and the expression of caspase-3 were determined using flow cytometry. RESULTS： Treatment with rAd-p53 or OXA alone inhibited gastric cancer cell growth in a time- and dose- dependent manner; moreover, significant synergistic effects were observed when these treatments were combined. Immunohistochemical analysis demonstrated that treatment with rAd-p53 alone, OXA alone or combined treatment led to decreased Bcl-2 expression and increased Bax expression in gastric cancer cells.Furthermore, flow cytometry showed that rAd-p53 alone, OXA alone or combination treatment induced apoptosis of gastric cancer cells, which was accompa- nied by increased expression of caspase-3. CONCLUSION： rAd-p53 enhances the sensitivity of gastric cancer cells to chemotherapy by promoting apoptosis. Thus, our results suggest that p53 gene therapy combined with chemotherapy represents a novel avenue for gastric cancer treatment.
基金Supported by the Chinese National Natural Scientific Fund(30472226)
文摘Objective:To address how genistein sensitizes the chemotherapy-resistant ovarian carcinoma cells and promotes apoptosis in the respect of cell cycle and the regulation of survivin expression in the process.Methods:Ovarian SKOV-3 carcinoma cell line was treated with genistein or cisplatin either alone or in combination.Cell viability was showed by MTT method.Cell cycle and apoptosis were detected by flow cytometry.Survivin mRNA and protein were revealed by RT-PCR and immunocytochemistry,respectively.Results:Genistein could reduce the cell viability in a dose-dependent manner,while cisplatin did so at a much higher level.In contrast,if the two agents were treated in combination,half growth inhibition(IC50) value for cisplatin was reduced remarkably and the effect was synergistic as analyzed by isobologram.In particular,the reduced cell viability was exhibited by a switch in cell cycle progression,as the cells were arrested in G2/M phase and the G0/G1 phase-fraction was significantly decreased.The reduced cell viability appeared to involve apoptosis,based on our results from flow cytometry and Hoechst 33258 staining.In the meanwhile,genistein performed the inhibitory effect on cisplatin-induced survivin expression.Conclusion:Genistein can sensitize ovarian carcinoma cells to cisplatin therapy with the inhibition of survivin expression as the potential mechanism.
基金Research Foundation of Guangdong Province (No. 99-Z-030-01)
文摘Objective: To investigate the association of the transportation characteristics of nolatrexed in tumor cells with its drug sensitivity. Methods: The sensitivity of 3 tumor cell lines, C6, SRS82 and LoVo, to nolatrexed were determined by growth inhibition study. After exposure to 20 μmol/L nolatrexed at different time intervals ranging from 0 to 30 min, or to nolatrexed at different concentrations ranging from 0 to 40 μmol/L for 10 min, the intracellular drug concentration was measured using high-performance liquid chro-matography. Results: C6 was the most sensitive cell line among the three, with sensitivity 6. 8-fold and 13. 8-fold those of SRS-82 and LoVo cells respectively. Transportation of nolatrexed in the 3 cell lines were qualitatively similar, which rapidly achieved steady-state within 5 min, and linear relationship between the intracellular and extracellular drug concentration was observed. The intracellular steady-state level achieved in C6 was significantly higher than those in the other two cell lines, the latter having comparable levels. Conclusion: Nolatrexed enters the cell very quickly and different transport capacities are involved in the generation of varied sensitivity to nolatrexed in tumor cells.
基金Supported by grants of foundation of "973" Program (No. 2009CB521802)National Natural Science foundation of China (No. 30872472,30973496,30800569)Natural Science Foundation of HuBei Province (No. 2008CDB174,2009CDB239)
文摘Objective:The aim of this study was to investigate the sensitivity of chemotherapeutic agents 5-FU,cisplatin(DDP) or TAXOL on colon cancer cell line SW480 with different methods,to find out the best examine time period for further study of chemotherapeutic sensitivities.Methods:The SW480 cell was treated with 5-FU(200μg/mL),DDP(150μg/mL) or TAXOL(50μg/mL) respectively for 4h,8h or 12h.MTT assay was used to examine the cell survival rate,Annexin V/PI assay was used to analysis the apoptosis rate,Western Blot assay was applied to examine the expression of apoptotic protein.Results:(1) Results of MTT assay showed that the survival rate of SW480 cells at 4h,8h or 12h was:5-FU(200μg/mL)96.0%±8.2%,85.4%±7.8%,74.4% ±10.2%(P<0.05);DDP(150μg/mL) 99.0%±6.4%,88.7%±4.7%(P<0.05),46.9%±2.6%(P<0.01);TAXOL(50μg/mL) 51.5%±4.2%(P<0.01),31.9%±3.1%,17.6%±2.3%,or blank group 97.2%±5.8%,98.7%±7.2%,97.5%±7.5% respectively.(2) The apoptosis rate of cancer cells at 4h,8h or 12h was:control group:3.4%±0.2%,6.2%±0.4%,7.0%±0.5%;5-FU(200μg/mL) 4.0%±0.3%,4.8%±0.4%,7.7%±0.7%;DDP(150μg/mL) 8.5%±0.9%,18.6%±1.6%(P<0.05),67.0%±6.2%(P<0.01);or TAXOL(50μg/mL) 32.0%±5.2%(P<0.01),76.6%±8.5%,94.0% ±8.2%,respectively.(3) Western Blot assay showed that the expression of apoptosis associated protein.PARP,X-linked inhibitor of apoptasis(XIAP),Caspase-9 and Bcl-xL were changed.Conclusion:The sensitivity of chemotherapy could be assessed by MTT assay,Annexin V/PI assay and Western Blot.The best examine time of the three chemotherapuc agents was 5-FU(200μg/mL):>12h,DDP(150μg/mL):8-12h,or TAXOL(50μg/mL):<4h.
文摘OBJECTIVE To explore the chemotherapeutic susceptibility of SP cells in human pulmonary adenocarcinoma cell line A549 and the possible mechanism of multidrug resistance.METHODS SP and non-SP （NSP） cells in the cell line A549 were isolated by fluorescence activated cell sorter. The susceptibility of SP and NSP cells to DDP, 5-FU, VP16, NVB and GEM was detected by a drug susceptibility test, and IC50s were calculated 24 h a er the chemotherapy; and then a er a 2-hour IC50 treatment with 5 chemotherapeutic drugs on the 2 subsets of NSP cells, the intracellular drug levels were determined and analyzed using high performance liquid chromatograph.RESULTS There was no statistical signifi cance in comparison of the di. erences in IC50s and in intracellular drug levels a er DDP treatment between the 2 subsets （P 〉 0.05）, （P 〉 0.05）. However,all IC50s of the other 4 drugs were significantly higher in the SP cells than in the NSP cells （P 〈 0.01）. A er the chemotherapy, the intracellular drug levels of the other 4 drugs were significantly lower in SP cells than in NSP cells （P 〈 0.01）.CONCLUSION Compared to NSP cells, SP cells in the cell line A549 have stronger resistance to the chemotherapeutics. The multidrug resistance of SP cells closely correlates with the function of SP cells discharging chemotherapeutic agents.
基金supported by National Natural Science Foundation of China(81102459)the Fundamental Research Funds for the Central Universities,Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources(Guangxi Normal University)+1 种基金Ministry of Education of China(CMEMR2012-B07)Doctoral Fund of Ministry of Education of China(20130071110069)
文摘K-ras is a member of ras gene family which is involved in cell survival,proliferation and differentiation.When a mutation occurs in ras gene,the activation of Ras proteins may be prolonged to induce oncogenesis.However,the relationship between K-ras mutation and clinical outcomes in pancreatic cancer patients treated with chemotherapy agents is still under debate.In this study,we constructed five pAcGFP1-C3 plasmids for different types of K-ras gene(WT,G12V,G12R,G12D,and G13D)and stably transfected human pancreatic cancer Bxpc-3 cells with these genes.The wild type and mutant clones showed a comparable growth and expression of K-Ras-GFP fusion protein.The expression of some K-ras mutations resulted in a reduced sensitivity to gefitinib,5-FU,docetaxel and gemcitabine,while showed no effects on erlotinib or cisplatin.Moreover,compared with the wild type clone,K-Ras downstream signals(phospho-Akt and/or phospho-Erk)were increased in K-ras mutant clones.Interestingly,different types of K-ras mutation had non-identical K-Ras downstream signal activities and drug responses.Our results are the first to reveal the relationship between different K-ras mutation and drug sensitivities of these anti-cancer drugs in pancreatic cancer cells in vitro.
