人MT1H 基因对肺腺癌细胞A549 增殖的影响及其机制

 目的 研究外源性人金属硫蛋白1H（MT1H）基因稳定转染对肺腺癌细胞A549生长的影响及其机制。方法 构建MT1H基因真核表达质粒pcDNA3．1（-）-MTIH，以脂质体法转染A549细胞，经G418筛选，获得阳性单克隆细胞。用RT-PCR和Western blot技术检测转染前后MT1H mRNA和蛋白表达，MTT实验检测其增殖能力的变化，流式细胞仪分析细胞周期时相分布。然后RT-PCR检测cy-clinD1的表达。结果 MT1H在A549细胞中获得表达。与未转染组和空质粒转染组比较，pcDNA3．1（一）～MT1H转染组细胞增殖速度明显增快，G0／G1期细胞减少，S期细胞增多，cyclinD1 mRNA水平明显升高。结论 MT1H能够促进肺腺癌细胞A549增殖，可能和上调cyclinD1促使细胞进入S期增多有关。

The Effect of Human MT1H Gene on the Proliferation in Lung Adenocarcinoma Cell A549 and Its Mechanism

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 protein were expressed in A549 cells.Cells tranfected pcDNA3.1(-)-MT1H compared with the control groups,proliferation rate significantly enhanced,cells number of G0/Gl phase obviously decreased while cells number of S phase increased,cyclin D1 mRNA levels elevated.Conclusion MT1H could promote proliferation of lung adenocarcinoma cell A549 by up-regulating expression of cyclin D1 to facilitate cells into S phase.