In recent years,the immune-modulatory role of all-trans astaxanthin from different pigment sources has been studied.It was reported that all-trans astaxanthin might exist as three stereoisomers,and the composition of ...In recent years,the immune-modulatory role of all-trans astaxanthin from different pigment sources has been studied.It was reported that all-trans astaxanthin might exist as three stereoisomers,and the composition of all-trans stereoisomers in natural materials differs from that of synthetic products.However,the different biological effects of various all-trans stereoisomers still remain unclear.In the present study,we evaluated the bioactivity of three astaxanthin stereoisomers,(3S,3'S)-trans-,(3R,3'R)-transand meso-trans-astaxanthin,in regulating cell-mediated immune response using mice lymphocytes and peritoneal exudates cells(PECs) systems.After the treatment with three astaxanthin stereoisomers(20 μmol L-1),the lymphocyte proliferation capacity,neutral red phagocytosis of PECs and natural killer(NK) cell cytotoxic activity were comparatively assessed.The results showed that all three astaxanthin stereoisomers significantly promoted lymphocyte proliferation,phagocytic capacity of PECs,and cytotoxic activity of NK cells.Moreover,the(3S,3'S)-trans-astaxanthin exhibited a much higher response than others.展开更多
OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxycycline were studied in mice transplanted with B16 ...OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxycycline were studied in mice transplanted with B16 melanoma cells. The mice were divided into 4 groups that were treated as follows： endostatin treatment （E group）, doxycycline treatment （D group）, endostatin plus doxycycline trearment （DE group）, controls （C group） received no treatment. Following 9 days of treatment the tumor tissue was removed to compare the differences in the tumor necrotic rate and micro-vessel density （MVD） among the different groups. Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen （PCNA） in the different groups.RESULTS The MVD of the 3 experimental groups was significantly less than the control group, （F = 10.888, P 〈 0.05）, indicating that doxycycline and endostatin can inhibit tumor angiogenesis by decreasing the tumor blood supply. This effect results in inhibition of tumor cellular proliferation and promotion of tumor cell necrosis. The tumor cell necrotic rate of the 3 experimental groups were all significantly higher than the C group （F = 7.229, P 〈 0.05） and the difference between the DE and C groups also was statistically significant. PCNA expression in all 3 experimental groups was statistically less than the C group （F = 17.729, P 〈 0.05）.CONCLUSION The combined use of endostatin and doxycycline in vivo can influence PCNA expression and angiogenesis in melanoma, and significantly inhibit melanoma cellular proliferation.展开更多
OBJECTIVE Hepatocyte growth factor （HGF） expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration o...OBJECTIVE Hepatocyte growth factor （HGF） expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing HGF expression. METHODS HGF was silenced using specific HGF a/f3 siRNA. The proliferation, migration, cell cycle and ultrastructure of SW620 cells were examined. RESULTS The transfection efficiency was 70%-80%. The expression rate of HGF in the experimental group was significantly lower than that in the negative and blank control groups （P 〈 0.05）. The proliferation inhibition rate in the experimental group at 24, 48, 72 and 96 h after transfection was 14.2%, 50.2%, 39.5% and 23.2%, respectively. The migratory ability of cells in the experimental group was significantly inhibited compared with that in the negative control or blank control groups （58.2% vs. 2.1% or 0%, P 〈 0.05）. CONCLUSION The application of RNA interference to silence the expression of HGF in the colon carcinoma cell line SW620 effectively inhibits the proliferation and migration of tumor cells.展开更多
Objective:The aim of the study was to investigate the biological character change of breast cancer cell line by inhibiting the Cyclin E expressing level.Methods:Human breast cancer cell line MCF-7 was transfected by C...Objective:The aim of the study was to investigate the biological character change of breast cancer cell line by inhibiting the Cyclin E expressing level.Methods:Human breast cancer cell line MCF-7 was transfected by Cyclin E siRNA vector pEGFP/CCNE2.SiRNA-induced silencing of Cyclin E was determined at RNA level and protein level,respectively.The proliferation of MCF-7 and its sensitivity to chemical therapy were measured by CCK-8 assay,while cell cycle was measured by flow cytometry(FCM).Results:The plasmid could reduce the expression of Cyclin E.The proliferation ability of MCF-7 was decreased and the sensitivity to chemical therapy was enhanced according to the inhibition of Cyclin E expression.The transfected MCF-7 was arrested at G1 phase of cell cycle.Conclusion:The inhibition of Cyclin E can decrease the breast cancer cell's growth,increase its sensation to chemical therapy and slow down its cell cycle.The Cyclin E siRNA may provide us a practical tool for further study on the gene therapy to breast cancers.展开更多
OBJECTIVE To explore the effect of photodynamic therapy with benzoporphyrin derivative monoacid ring A （BPD-MA） on the proliferation and apoptosis of human bladder cancer cells. METHODS Photosensitization of BPD-MA ...OBJECTIVE To explore the effect of photodynamic therapy with benzoporphyrin derivative monoacid ring A （BPD-MA） on the proliferation and apoptosis of human bladder cancer cells. METHODS Photosensitization of BPD-MA was activated with a red light laser （632.8 nm） delivered at 10 mw/cm^2 to give a total dose of 2.4 J/cm^2. Cellular proliferative activity was measured using the 3-（4,5.- dimethylethiazil-2-yl）-2,5-Diph3-eyl tetrazolium bromide （MFi-） assay and 3H-thymidine incorporation. Cell apoptosis was determined with flow cytometry analysis and the terminal deoxyuridine nicked-labeling （TUNEL） assay. RESULTS At 24 h post photodynamic treatment, photodynamic therapy significantly decreased cellular proliferative activity. The rate of apoptosis in BIU-87 cells 8 h after photodynamic treatment significantly increased up to 26.11± 2.59% as analyzed with flow cytometry. In situ labeling of DNA cleavage products with the terminal deoxyuridine nicked-labeling （TUNEL） assay reinforced these observations, BPD-MA-mediated photosensitization increased the number of TUNEL-positive cells compared to the controls. However, laser irradiation alone, BPD-MA alone and sham radiation did not affect cellular proliferative activity or apoptosis of the human bladder cancer BIU-87 cells. CONCLUSION Photodynamic therapy with BPD-MA significantly decreases cellular proliferative activity and enhances apoptosis. Therapy using this method might be a promising approach to treat patients with bladder cancer.展开更多
The MTEC1 cell line, established in our laboratory, is a normal epithelial cell line derived from thymus medulla of Balb/c mice and these cells constitutively produce multiple cytokines. The selection of thymic microe...The MTEC1 cell line, established in our laboratory, is a normal epithelial cell line derived from thymus medulla of Balb/c mice and these cells constitutively produce multiple cytokines. The selection of thymic microenvironment on developing T cells was investigated in an in vitro system. Un-separated fresh thymocytes from Balb/c mice were cocul-tured with MTECl cells or/and MTEC1-SN,then,the viability, proliferation and phenotypes of cultured thymocytes were assessed. Without any exogenous stimulus, both MTECl cells and MTECl -SN were able to maintain the viability of thymocytes, while only the MTECl cells, not the MTECl -SN, could directly activate thymocytes to exhibit moderate proliferation, indicating that the proliferative signal is delivered through cell surface interactions of MTECl cells and thymocytes. Phenotype analysis on FACS of viable thymocytes after coculture revealed that MTECl cells preferentially activate the subsets of CD4+ CDS', CD4+ CD8+ and CD4- CD8- thymocytes; whereas MTEC1- SN preferentially maintained the viability of CD4+ CD8- and CD4-CD8+ thymocyte subsets.For the Con A-activated thymocytes, both MTEC1 cells and MTEC1-SN provided accessory signal(s) to significantly increase the number of viable cells and to markedly enhance the proliferation of thymocytes with virtually equal potency, phenotyped as CD4+CD8-, CD4-CD8+, and CD4- CD8- subsets. In summary, MTEC1 cells displayedselective support to the different thymocyte subsets , and the selectivity is dependent on the status of thymocytes.展开更多
OBJECTIVE: To examine whether a combinative treatment with curcumin enhances the effects of the epidermal growth factor receptor-tyrosine kinase inhibitor(EGFR-TKI) gefitinib on cell proliferation, clonogenic capacity...OBJECTIVE: To examine whether a combinative treatment with curcumin enhances the effects of the epidermal growth factor receptor-tyrosine kinase inhibitor(EGFR-TKI) gefitinib on cell proliferation, clonogenic capacity and apoptosis in the drug-resistant lung cancer cell line NCI-H1975, and further investigate the molecular mechanisms involved.METHODS: NCI-H1975 cells were treated with curcumin and gefitinib alone or in combination, and cell proliferation, clonogenic capacity and apoptosis were examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, clone forming experiments, and flow cytometry, respectively, while p38, extracellular regulated protein kinase(ERK)1/2, and protein kinase B(AKT)phosphorylation were examined using Western blotting.RESULTS: Compared with the effects of either agent alone, the combination of curcumin and gefitinib had a stronger suppressive effect on proliferation and the clonogenic capacity(P < 0.05), and showed an increased ability to promote apoptosis(P < 0.05) and reduce p38, ERK1/2, and AKT phosphorylation(P < 0.05).CONCLUSION: Co-treatment of curcumin and gefitinib significantly improves the ability of gefitinib to inhibit cell proliferation, suppress the clonogenic capacity and enhance apoptosis in NCI-H1975 cells,and these effects are possibly mediated via a decrease in phosphorylation of proteins in downstream pathways of the epidermal growth factor receptor.展开更多
p75NTR is a low-affinity nerve growth factor receptor, which promotes cell proliferation as a positive modulator of high-affinity receptor TrkA, as well as binds with cell ligands to induce apoptosis and mediate death...p75NTR is a low-affinity nerve growth factor receptor, which promotes cell proliferation as a positive modulator of high-affinity receptor TrkA, as well as binds with cell ligands to induce apoptosis and mediate death signals. To analyze the regulatory mechanisms of p75NTR, the present study utilized a new membrane yeast two-hybrid system to screen a human fe- tal brain cDNA library. Results identified BFAR, a novel protein that interacts with p75NTR. Interaction specificity was veri- fied by membrane yeast two-hybrid co-transformation assays, in vitro GST pull-down assays, and in vitro co-irnmunopreci- pitation assays. The fluorescent subcellular localization assay revealed that the two proteins co-localized within the cytoplasm. BFAR overexpression in PC-12 and HEK293T cells inhibited the NFnB and JNK signaling pathway, as determined with the luciferase test. Co-transfected p75NTR and BFAR in HEK293T or PC-12 cells, respectively, increased the percentage of cells in the G2/M phase, decreased the number of S-phase cells, and did not change the number of G0/Gl-phase cells.展开更多
Olfactory ensheathing cells(OECs) transplanted into the damaged spinal cord may be considered as a valuable remedy explorations for spinal cord repair. The proliferation of transplanted olfactory ensheathing cells dep...Olfactory ensheathing cells(OECs) transplanted into the damaged spinal cord may be considered as a valuable remedy explorations for spinal cord repair. The proliferation of transplanted olfactory ensheathing cells depends on various environmental factors and effective cues, which may include electrical fields(EFs). In this study, we investigated the proliferative capacity, morphologic alterations of olfactory ensheathing cells derived from neonate rat that occurd when exposed to two EFs of 20 Hz, 50 mV and20 Hz, 100 mV for 6 h. For both EF treatments, the MTT results revealed that the cellular proliferation of exposed group during the last 6 h of the experiment was statistically higher than that of control group. Then, we investigated morphological structure changes in the cells stained by Coomassie brilliant blue. Compared with control group, most of cells were present at intensively proliferating appearance including the microfilaments were long and thick and the accumulated appearance of cells. It is conceivable that electrical fields as a new approach may promote the growth and proliferation of OECs and may be engineered to control the survival of transplanted OECs in injured spinal cord.Although our results have been suggesting that EFs may be non-chemical strategies for cell proliferation, the fundamental mechanisms remain to be elucidated.展开更多
基金supported by Program for Changjiang Scholars and Innovative Research Team in University (IRT1188)
文摘In recent years,the immune-modulatory role of all-trans astaxanthin from different pigment sources has been studied.It was reported that all-trans astaxanthin might exist as three stereoisomers,and the composition of all-trans stereoisomers in natural materials differs from that of synthetic products.However,the different biological effects of various all-trans stereoisomers still remain unclear.In the present study,we evaluated the bioactivity of three astaxanthin stereoisomers,(3S,3'S)-trans-,(3R,3'R)-transand meso-trans-astaxanthin,in regulating cell-mediated immune response using mice lymphocytes and peritoneal exudates cells(PECs) systems.After the treatment with three astaxanthin stereoisomers(20 μmol L-1),the lymphocyte proliferation capacity,neutral red phagocytosis of PECs and natural killer(NK) cell cytotoxic activity were comparatively assessed.The results showed that all three astaxanthin stereoisomers significantly promoted lymphocyte proliferation,phagocytic capacity of PECs,and cytotoxic activity of NK cells.Moreover,the(3S,3'S)-trans-astaxanthin exhibited a much higher response than others.
基金a grant from National Natural Science Foundation of China (No.30370554)
文摘OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxycycline were studied in mice transplanted with B16 melanoma cells. The mice were divided into 4 groups that were treated as follows： endostatin treatment （E group）, doxycycline treatment （D group）, endostatin plus doxycycline trearment （DE group）, controls （C group） received no treatment. Following 9 days of treatment the tumor tissue was removed to compare the differences in the tumor necrotic rate and micro-vessel density （MVD） among the different groups. Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen （PCNA） in the different groups.RESULTS The MVD of the 3 experimental groups was significantly less than the control group, （F = 10.888, P 〈 0.05）, indicating that doxycycline and endostatin can inhibit tumor angiogenesis by decreasing the tumor blood supply. This effect results in inhibition of tumor cellular proliferation and promotion of tumor cell necrosis. The tumor cell necrotic rate of the 3 experimental groups were all significantly higher than the C group （F = 7.229, P 〈 0.05） and the difference between the DE and C groups also was statistically significant. PCNA expression in all 3 experimental groups was statistically less than the C group （F = 17.729, P 〈 0.05）.CONCLUSION The combined use of endostatin and doxycycline in vivo can influence PCNA expression and angiogenesis in melanoma, and significantly inhibit melanoma cellular proliferation.
基金This work was supported by a grant from the Natural Science Foundation of Hebei Province of China （No.05547008D-3）.
文摘OBJECTIVE Hepatocyte growth factor （HGF） expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing HGF expression. METHODS HGF was silenced using specific HGF a/f3 siRNA. The proliferation, migration, cell cycle and ultrastructure of SW620 cells were examined. RESULTS The transfection efficiency was 70%-80%. The expression rate of HGF in the experimental group was significantly lower than that in the negative and blank control groups （P 〈 0.05）. The proliferation inhibition rate in the experimental group at 24, 48, 72 and 96 h after transfection was 14.2%, 50.2%, 39.5% and 23.2%, respectively. The migratory ability of cells in the experimental group was significantly inhibited compared with that in the negative control or blank control groups （58.2% vs. 2.1% or 0%, P 〈 0.05）. CONCLUSION The application of RNA interference to silence the expression of HGF in the colon carcinoma cell line SW620 effectively inhibits the proliferation and migration of tumor cells.
基金Supported by a grant of National Natural Science Foundation of China(No. 30801116)
文摘Objective:The aim of the study was to investigate the biological character change of breast cancer cell line by inhibiting the Cyclin E expressing level.Methods:Human breast cancer cell line MCF-7 was transfected by Cyclin E siRNA vector pEGFP/CCNE2.SiRNA-induced silencing of Cyclin E was determined at RNA level and protein level,respectively.The proliferation of MCF-7 and its sensitivity to chemical therapy were measured by CCK-8 assay,while cell cycle was measured by flow cytometry(FCM).Results:The plasmid could reduce the expression of Cyclin E.The proliferation ability of MCF-7 was decreased and the sensitivity to chemical therapy was enhanced according to the inhibition of Cyclin E expression.The transfected MCF-7 was arrested at G1 phase of cell cycle.Conclusion:The inhibition of Cyclin E can decrease the breast cancer cell's growth,increase its sensation to chemical therapy and slow down its cell cycle.The Cyclin E siRNA may provide us a practical tool for further study on the gene therapy to breast cancers.
文摘OBJECTIVE To explore the effect of photodynamic therapy with benzoporphyrin derivative monoacid ring A （BPD-MA） on the proliferation and apoptosis of human bladder cancer cells. METHODS Photosensitization of BPD-MA was activated with a red light laser （632.8 nm） delivered at 10 mw/cm^2 to give a total dose of 2.4 J/cm^2. Cellular proliferative activity was measured using the 3-（4,5.- dimethylethiazil-2-yl）-2,5-Diph3-eyl tetrazolium bromide （MFi-） assay and 3H-thymidine incorporation. Cell apoptosis was determined with flow cytometry analysis and the terminal deoxyuridine nicked-labeling （TUNEL） assay. RESULTS At 24 h post photodynamic treatment, photodynamic therapy significantly decreased cellular proliferative activity. The rate of apoptosis in BIU-87 cells 8 h after photodynamic treatment significantly increased up to 26.11± 2.59% as analyzed with flow cytometry. In situ labeling of DNA cleavage products with the terminal deoxyuridine nicked-labeling （TUNEL） assay reinforced these observations, BPD-MA-mediated photosensitization increased the number of TUNEL-positive cells compared to the controls. However, laser irradiation alone, BPD-MA alone and sham radiation did not affect cellular proliferative activity or apoptosis of the human bladder cancer BIU-87 cells. CONCLUSION Photodynamic therapy with BPD-MA significantly decreases cellular proliferative activity and enhances apoptosis. Therapy using this method might be a promising approach to treat patients with bladder cancer.
基金grants from the China medical board, USA,and from the national foundation of natural scirnces,China
文摘The MTEC1 cell line, established in our laboratory, is a normal epithelial cell line derived from thymus medulla of Balb/c mice and these cells constitutively produce multiple cytokines. The selection of thymic microenvironment on developing T cells was investigated in an in vitro system. Un-separated fresh thymocytes from Balb/c mice were cocul-tured with MTECl cells or/and MTEC1-SN,then,the viability, proliferation and phenotypes of cultured thymocytes were assessed. Without any exogenous stimulus, both MTECl cells and MTECl -SN were able to maintain the viability of thymocytes, while only the MTECl cells, not the MTECl -SN, could directly activate thymocytes to exhibit moderate proliferation, indicating that the proliferative signal is delivered through cell surface interactions of MTECl cells and thymocytes. Phenotype analysis on FACS of viable thymocytes after coculture revealed that MTECl cells preferentially activate the subsets of CD4+ CDS', CD4+ CD8+ and CD4- CD8- thymocytes; whereas MTEC1- SN preferentially maintained the viability of CD4+ CD8- and CD4-CD8+ thymocyte subsets.For the Con A-activated thymocytes, both MTEC1 cells and MTEC1-SN provided accessory signal(s) to significantly increase the number of viable cells and to markedly enhance the proliferation of thymocytes with virtually equal potency, phenotyped as CD4+CD8-, CD4-CD8+, and CD4- CD8- subsets. In summary, MTEC1 cells displayedselective support to the different thymocyte subsets , and the selectivity is dependent on the status of thymocytes.
基金Supported by Natural Science Found of Zhejiang Province:Research of Molecular Mechanism of Curcumin Reversing TKI Targeted Drug Resistance of NSCLC(No.LY13H160037)
文摘OBJECTIVE: To examine whether a combinative treatment with curcumin enhances the effects of the epidermal growth factor receptor-tyrosine kinase inhibitor(EGFR-TKI) gefitinib on cell proliferation, clonogenic capacity and apoptosis in the drug-resistant lung cancer cell line NCI-H1975, and further investigate the molecular mechanisms involved.METHODS: NCI-H1975 cells were treated with curcumin and gefitinib alone or in combination, and cell proliferation, clonogenic capacity and apoptosis were examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, clone forming experiments, and flow cytometry, respectively, while p38, extracellular regulated protein kinase(ERK)1/2, and protein kinase B(AKT)phosphorylation were examined using Western blotting.RESULTS: Compared with the effects of either agent alone, the combination of curcumin and gefitinib had a stronger suppressive effect on proliferation and the clonogenic capacity(P < 0.05), and showed an increased ability to promote apoptosis(P < 0.05) and reduce p38, ERK1/2, and AKT phosphorylation(P < 0.05).CONCLUSION: Co-treatment of curcumin and gefitinib significantly improves the ability of gefitinib to inhibit cell proliferation, suppress the clonogenic capacity and enhance apoptosis in NCI-H1975 cells,and these effects are possibly mediated via a decrease in phosphorylation of proteins in downstream pathways of the epidermal growth factor receptor.
基金supported by the National High Technology Research and Development Program of China (Grant No. 2006AA02A310)National Science and Technology Key Program of China (Grant Nos. 2008ZX10003-006 and 2009ZX09301011)National Basic Research Program of China (Grant No. 2010CB912609) which were awarded to Huo KeKe
文摘p75NTR is a low-affinity nerve growth factor receptor, which promotes cell proliferation as a positive modulator of high-affinity receptor TrkA, as well as binds with cell ligands to induce apoptosis and mediate death signals. To analyze the regulatory mechanisms of p75NTR, the present study utilized a new membrane yeast two-hybrid system to screen a human fe- tal brain cDNA library. Results identified BFAR, a novel protein that interacts with p75NTR. Interaction specificity was veri- fied by membrane yeast two-hybrid co-transformation assays, in vitro GST pull-down assays, and in vitro co-irnmunopreci- pitation assays. The fluorescent subcellular localization assay revealed that the two proteins co-localized within the cytoplasm. BFAR overexpression in PC-12 and HEK293T cells inhibited the NFnB and JNK signaling pathway, as determined with the luciferase test. Co-transfected p75NTR and BFAR in HEK293T or PC-12 cells, respectively, increased the percentage of cells in the G2/M phase, decreased the number of S-phase cells, and did not change the number of G0/Gl-phase cells.
文摘Olfactory ensheathing cells(OECs) transplanted into the damaged spinal cord may be considered as a valuable remedy explorations for spinal cord repair. The proliferation of transplanted olfactory ensheathing cells depends on various environmental factors and effective cues, which may include electrical fields(EFs). In this study, we investigated the proliferative capacity, morphologic alterations of olfactory ensheathing cells derived from neonate rat that occurd when exposed to two EFs of 20 Hz, 50 mV and20 Hz, 100 mV for 6 h. For both EF treatments, the MTT results revealed that the cellular proliferation of exposed group during the last 6 h of the experiment was statistically higher than that of control group. Then, we investigated morphological structure changes in the cells stained by Coomassie brilliant blue. Compared with control group, most of cells were present at intensively proliferating appearance including the microfilaments were long and thick and the accumulated appearance of cells. It is conceivable that electrical fields as a new approach may promote the growth and proliferation of OECs and may be engineered to control the survival of transplanted OECs in injured spinal cord.Although our results have been suggesting that EFs may be non-chemical strategies for cell proliferation, the fundamental mechanisms remain to be elucidated.
