MT1H gene on human A549 lung adenocarcinoma cell proliferation and its mechanism.doc

 PAGE \* MERGEFORMAT 14 MT1H gene on human A549 lung adenocarcinoma cell proliferation and its mechanism Author: Hou Xin-Fang, Fan Qingxia, Wang left Xing, Wang Ruilin, Roth Xin, Zhao Peirong 【Abstract】 Objective To study the exogenous human metallothionein 1H (MT1H) gene stable transfection of A549 lung adenocarcinoma cell growth and its mechanism. Methods to build MT1H eukaryotic expression plasmid pcDNA3.1 ()  MT1H, with liposome transfected A549 cells by G418 selection, access to positive monoclonal cell. By RT  PCR and Western blot were detected before and after transfection MT1H mRNA and protein expression, MTT proliferation test of its ability to detect changes in flow cytometry analysis of cell cycle phase distribution. Then RT  PCR detection of cyclinD1 expression. Results MT1H in A549 cells expressed. With non-transfected group and empty vector transfected group, pcDNA3.1 ()  MT1H transfection group increased significantly faster rate of cell proliferation, G0/G1 phase cells decreased, S-phase cells increased, cyclinD1 mRNA levels were significantly elevated . Conclusion MT1H can promote the proliferation of A549 lung adenocarcinoma cells, may be raised cyclinD1 prompted an increase in the cell to enter S phase. Keywords: lung cancer; Metallothionein 1H; cell proliferation; cyclinD1 Abstract: Objective To investigate the effects of exogenous MT1H gene stably transfection on growth of lung adenocarcinoma cell A549 in vitro. Methods A recombinant eukaryotic expression plasmid pcDNA3.1 ()  MT1H was constructed and transfected into A549 cells by lipofectamine 2000. Positive monoclone was screened by G418. RT  PCR and Western blot assays were used to identify the expression of MT1H mRNA and protein respectively. The cell proliferation was determined by MTT assay. The alterations of cell cycle were determined by flow cytometry (FCM). Then the expression of cyclinD1 mRNA was determined by RT  PCR. Results MT1H mRNA and