金属硫蛋白1E对胰腺癌SW1990细胞生长的影响

目的 观察外源性人金属硫蛋白1E(MT1E)基因对胰腺癌SW1990细胞生长的影响.方法 构建MT1E基因真核表达载体,转染SW1990细胞并获得阳性单克隆细胞.用Westernblot技术检测转染前后MT1E蛋白表达,噻唑蓝(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.同时检测Cyclin D1与p53的表达.结果 MT1E融合蛋白在SW1990MT1E细胞中表达.MTT结果表明,与SW1990和SW1990MT1E空细胞比较,SW1990MT1E细胞增殖速度明显增快.SW1990MT1E细胞与SW1990和SW1990空细胞比较,G0/G1期细胞明显减少,S期明显增多,差异有统计学意义.SW1990MT1E较SW1990空的细胞Cyclin D1与p53表达明显升高.结论 MT1E能够促进SW1990增殖,可能和上调Cyclin D1促使细胞进入S期增多,通过失活p53蛋白使细胞恶性变.

Effects of MT1E gene on proliferation in human pancreatic carcinoma cell line SW1990

Objective To investigate the effects of MT1E gene on the cell proliferation in human pancreatic carcinoma cell line SW1990.Methods Human pancreatic carcinoma cell line SW1990 made by transfection with expressible MT1E gene, Positive monoclone was screened by G418.Western blotting assays were used to identify the expression of MT1E protein respectively. The cell proliferation was determined by MTT assay. The cell cycle was detected by flow cytometry, The apoptosis-related protein levels of Cyclin Dl and p53 determined by immunohistochemistry. Results MT1E protein were expressed in SW1990.Cells tranfected pcDNA3.1 ( - ) -MT1E compared with the SW1990 and cells tranfected pcDNA 3.1 ( - ), proliferation rate significantly enhanced, cells number of G0/G1 phase obviously decreased while cells number of S phase increased, Cyclin Dl and p53 protein levels elevated. Conclusion MT1E could promote proliferation of SW1990 cell by upregulating expression of cyclin Dl and p53 to facilitate cells into S phase.