pcDNA3.1-CHD5表达载体构建及其表达对食管癌细胞生物学活性影响  

Effect of pc DNA3 1-CHD5 expression on the proliferation and metastasis of ECA 109 cells

作　　者：丛德刚[1] 

机构地区：[1]杭州师范大学附属医院胸外科,浙江杭州310015

出　　处：《健康研究》2015年第6期621-623,F0003,共4页Health Research

基　　金：杭州市科技发展计划项目(20120633B16)

摘　　要：目的研究CHD5基因表达对食管癌细胞系ECA109增殖和迁移的影响。方法用分子生物学方法构建pc DNA3.1-CHD5重组质粒,用重组质粒转染食管癌细胞系ECA109,RT-PCR方法检测ECA109中CHD5mRNA的表达,Western blot分析转染后CHD5蛋白表达情况。CCK8和细胞划痕实验检测pc DNA3.1-CHD5对ECA109细胞增殖的影响。结果 WB结果显示重组质粒处理后CHD5蛋白表达量明显上调。细胞划痕实验显示转染48h后,细胞增殖速度明显较对照组慢。结论体外实验显示CHD5基因的表达能够抑制ECA109细胞的增殖和迁移,并具有明显的时-效关系。验证了CHD5在食管鳞癌中发挥的是抑癌基因的作用的观点,提示其作为食管癌早期诊断和预后检测的生物标记值得尝试。Objective To examine the way in which the proliferation and metastasis of ECA109 cells are regulated by the expression of CHDS. Method We firstly constructed pcDNA3 1-CHD5 recombinant plasmid and then had it transfeeted with ECA109 esophageal cancer cell line. After transfection of pcDNA3 1-CHD5, CHD5 mRNA expression was tested by RT- PCR method. The protein expression was tested by Western blot. CCK8 and cell scratch test were used to test the proliferation of ECA109 cell. Findings Increased expression of CHD5 protein was observed after transfection. With cell scratch assay we found that the proliferation ratio of ECA109 cell was down-regulated 48 hours after transfecti0n. Conclusion Our data showed that CHD5 could inhibit the proliferative and invasive abilities of ECA 109 cells. It could successfully build pcDNA3 1-CHD5 plasmid and realized CHD5 gene expression level in ECA109 cells. Recombinant plasmid can inhibit the proliferation of ECA109 cells.

关 键 词：CHD5 食管癌细胞 真核表达质粒 

分 类 号：R36[医药卫生—病理学]