Objective:To induce pluripotent stem(IPS)cells from fibrocytes that are separated from liver cancer patients.Methods:The fibrocytes were reprogrammed to IPS cells by lentiviral vector,stained and identified by immunoh...Objective:To induce pluripotent stem(IPS)cells from fibrocytes that are separated from liver cancer patients.Methods:The fibrocytes were reprogrammed to IPS cells by lentiviral vector,stained and identified by immunohistochemistry.Results:The IPS cells were successfully established from fibrocytes after infection,and IPS cell clones formed in round shape under a microscopy.The induction rate was 0.013%±0.007%.No tumor formed at the back of nude mice within 8 weeks after the inoculation of cell clone.However,tetatoma appeared in nude mice within 1 week after IPS inoculation.A few tumors formed in nude mice within 4 weeks after the inoculation of cell clones.However,subcutaneous tumors formed within 1 week after IPS inoculation.The induced IPS cells showed three germ layers in tetatoma.Nanog and OCT4 in the induced IPS cells showed hypomethylation.SSEA-A,TRA-1-6-,TRA-1-81 and Nanog were highly expressed in the induced IPS cells,indicating the IPS cells possessed the similar ability as the stem cells.Conclusion:The IPS cells of liver cancer patients can be established effectively from fibrocytes and can be cultured stably in vitro,which provides an approach for the treatment of intermediate or advanced stage liver cancer.展开更多
Objective: To investigate the relationship between genetic factor and prostate cancer(Pca) risk and the possible cause in it. Methods: The polymorphisms of cytochrome P450 family 17(CYPl7) rs743572, p27 V109 G and and...Objective: To investigate the relationship between genetic factor and prostate cancer(Pca) risk and the possible cause in it. Methods: The polymorphisms of cytochrome P450 family 17(CYPl7) rs743572, p27 V109 G and androgen receptor(AR) gene CAG repeat length in peripheral blood from 70 cases and 70 controls were detected through the polymerase chain reaction-restriction fragment length polymorphism technique or short tandem repeatpolymerase chain reaction technique. Then, according to the results of case-control study, the recombinant plasmids containing the wild/mutant p27 gene were constructed and transfected Pca LNcap cells. After 24 and 72 h of transfection, the cell proliferative activity was determined by MTT method, cell cycle distribution and apoptosis was detected by flow cytometry, and the expression level of bcl-2, caspase-3 and p27 protein was determined by Western-blot. Results:In three target polymorphisms, only p27 V109 G polymorphism was related to Pca risk(P=0.030, OR=0.202, 95% CI=0.042-0.973). Pca risk of p27-109 G allele was lower than-109 V allele(P=0.006, OR=0.285, 95% CI=0.110-0.737). Cells transfected with wild/mutant p27 gene both showed the higher cells apoptosis rate and the lower cell proliferative activity than mock cells(P<0.05 or 0.01), the regulatory effect of mutant p27 on cell proliferation and apoptosis was stronger than the wild p27(P<0.05). Conclusions: p27-109 G allele that could cause higher p27 protein expression than-109 V allele in LNcap cells, maybe is the protective factor of Pca.展开更多
基金supported by Science and Technology Project of Henan Provincial Department of Health(NO:2011020038)
文摘Objective:To induce pluripotent stem(IPS)cells from fibrocytes that are separated from liver cancer patients.Methods:The fibrocytes were reprogrammed to IPS cells by lentiviral vector,stained and identified by immunohistochemistry.Results:The IPS cells were successfully established from fibrocytes after infection,and IPS cell clones formed in round shape under a microscopy.The induction rate was 0.013%±0.007%.No tumor formed at the back of nude mice within 8 weeks after the inoculation of cell clone.However,tetatoma appeared in nude mice within 1 week after IPS inoculation.A few tumors formed in nude mice within 4 weeks after the inoculation of cell clones.However,subcutaneous tumors formed within 1 week after IPS inoculation.The induced IPS cells showed three germ layers in tetatoma.Nanog and OCT4 in the induced IPS cells showed hypomethylation.SSEA-A,TRA-1-6-,TRA-1-81 and Nanog were highly expressed in the induced IPS cells,indicating the IPS cells possessed the similar ability as the stem cells.Conclusion:The IPS cells of liver cancer patients can be established effectively from fibrocytes and can be cultured stably in vitro,which provides an approach for the treatment of intermediate or advanced stage liver cancer.
基金supported by Science and Technology Plan Projects of Henan Province(132102310513)
文摘Objective: To investigate the relationship between genetic factor and prostate cancer(Pca) risk and the possible cause in it. Methods: The polymorphisms of cytochrome P450 family 17(CYPl7) rs743572, p27 V109 G and androgen receptor(AR) gene CAG repeat length in peripheral blood from 70 cases and 70 controls were detected through the polymerase chain reaction-restriction fragment length polymorphism technique or short tandem repeatpolymerase chain reaction technique. Then, according to the results of case-control study, the recombinant plasmids containing the wild/mutant p27 gene were constructed and transfected Pca LNcap cells. After 24 and 72 h of transfection, the cell proliferative activity was determined by MTT method, cell cycle distribution and apoptosis was detected by flow cytometry, and the expression level of bcl-2, caspase-3 and p27 protein was determined by Western-blot. Results:In three target polymorphisms, only p27 V109 G polymorphism was related to Pca risk(P=0.030, OR=0.202, 95% CI=0.042-0.973). Pca risk of p27-109 G allele was lower than-109 V allele(P=0.006, OR=0.285, 95% CI=0.110-0.737). Cells transfected with wild/mutant p27 gene both showed the higher cells apoptosis rate and the lower cell proliferative activity than mock cells(P<0.05 or 0.01), the regulatory effect of mutant p27 on cell proliferation and apoptosis was stronger than the wild p27(P<0.05). Conclusions: p27-109 G allele that could cause higher p27 protein expression than-109 V allele in LNcap cells, maybe is the protective factor of Pca.
