The effect of RASSF1A gene on the proliferation of human gastric carcinoma cell line SGC7901
Jifang Wen
Abstract
Jifang Wen
Abstract
Objective To study the effects of exogenous RASSF1A gene on the proliferation of gastric carcinoma cell line SGC7901.Methods The mammal expression vector pcDNA3.0-RASSF1A and pcDNA3.0 were introduced into SGC7901 cell line by lipofectin transfection,and the SGC7901 cells stably overexpressing RASSF1A gene were established by G418 selection.The expression of RASSF1A gene was detected by Western blotting and RT-PCR.The cytobiologic characterizations of the positive clone were analyzed by MTT assay,cytometry,colony assay,and tumorigenicity experiment.Results SGC7901 cells with stably expression RASSF1A protein were established by lipofection mediated transfection and selected for further study.Compared with the non-transfected and vector transfected cells,the positive clone cells developed more slowly;flow cytometric data showed that more positive clone cells went into phase G0/G1 and less cells went into phase S(P0.05);colony formation efficiency of the positive clone cells was lower than that of non-transfected and vector transfected cells(P0.05).The average weight of tumor tissue in nude mice in positive clone cells was less than that of non-transfected and vector transfected cells(P0.05).Conclusions RASSF1A gene can suppress the proliferation of SGC7901 cell line.
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Objective To study the effects of exogenous RASSF1A gene on the proliferation of gastric carcinoma cell line SGC7901.Methods The mammal expression vector pcDNA3.0-RASSF1A and pcDNA3.0 were introduced into SGC7901 cell line by lipofectin transfection,and the SGC7901 cells stably overexpressing RASSF1A gene were established by G418 selection.The expression of RASSF1A gene was detected by Western blotting and RT-PCR.The cytobiologic characterizations of the positive clone were analyzed by MTT assay,cytometry,colony assay,and tumorigenicity experiment.Results SGC7901 cells with stably expression RASSF1A protein were established by lipofection mediated transfection and selected for further study.Compared with the non-transfected and vector transfected cells,the positive clone cells developed more slowly;flow cytometric data showed that more positive clone cells went into phase G0/G1 and less cells went into phase S(P0.05);colony formation efficiency of the positive clone cells was lower than that of non-transfected and vector transfected cells(P0.05).The average weight of tumor tissue in nude mice in positive clone cells was less than that of non-transfected and vector transfected cells(P0.05).Conclusions RASSF1A gene can suppress the proliferation of SGC7901 cell line.
Key concepts: Transfection, clone (Java method), Molecular biology, Cell culture, Flow cytometry, Cell growth, Biology, Cell