DEC1基因真核表达质粒的构建及对胃癌细胞增殖的影响  

Effects of DEC1 on proliferation of gastric cancer cells:an approach through a eukaryotic over-expression vector

作　　者：戴新[1] 郑燕[2] 郏雁飞[2] 肖东杰[2] 童书青[1] 汪运山[2] 

机构地区：[1]山东大学,医学院,济南250012 [2]山东大学,附属济南市中心医院中心实验室,济南250013

出　　处：《山东大学学报（医学版）》2012年第6期75-79,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81000869);山东省优秀中青年科学家科研奖励基金(BS2011YY039);济南“泉城学者”建设工程招标课题(Q2-06)

摘　　要：目的构建人分化型胚胎软骨发育基因DEC1真核表达载体,探讨DEC1基因对胃癌细胞增殖的影响。方法提取人胃癌细胞MGC-803总RNA,RT-PCR扩增获得DEC1基因,将DEC1基因插入pGEM-T载体,再插入真核表达质粒pcDNA3,将成功构建的pcDNA3-DEC1载体介导转染MGC-803细胞。通过RT-PCR和Westernblot分别检测转染后细胞DEC1在mRNA和蛋白水平的表达;MTT法、免疫细胞化学法检测过表达DEC1基因后MGC-803细胞增殖的变化。结果转染pcDNA3-DEC1质粒的MGC-803细胞DEC1 mRNA和蛋白水平明显升高(P<0.05);MTT试验、ki-67免疫细胞化学染色显示转染后细胞增殖能力明显升高(P<0.05)。结论 DEC1的过表达能明显增强胃癌细胞MGC-803的增殖能力。Objective To construct a DEC1 eukaryotic expression vector and to observe the effects of the DEC1 gene on proliferation of MGC-803 cells （ a gastric cancer cell line）. Method The DEC1 gene was amplified by RT-PCR and cloned into pGEM-T, and then into the eukaryotic expression vector pcDNA3 resulting in a recombinant vector, named pcDNA3-DEC1, pcDNA3-DEC1 was transfected into MGC-803 cells. Expression of DEC1 was detected by RT- PCR and Western blot. The effect of DEC1 on cell proliferation was measured by MTF assay and immunocytochemis- try. Results After transfection of MGC-803 cells,their DEC1 expression at both the transcript and protein levels was significantly increased, corresponding to high rates of cell proliferation. Conclusion Expression of the DECI gene can promote cell proliferation of gastric cancer cells.

关 键 词：胃肿瘤 质粒构建 逆转录聚合酶链反应 转染 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]