军事医学2017，Vol.41Issue(10)：792-795,4.DOI:10.7644/j.issn.1674-9960.2017.10.002
重组pEGFP-C1-CXCL1真核表达载体构建及其对内质网应激下人肝癌细胞HepG2的增殖作用
Construction of pEGFP-C1-CXCL1 plasmid and detection of its expression in eukaryotic cells
摘要
Abstract
Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.关键词
趋化因子类/CXCL1/重组质粒/细胞增殖/癌,肝细胞Key words
chemotactic factors/CXCL1/recombinant plasmid/cell proliferation/carcinoma/hepatocellular分类
医药卫生引用本文复制引用
毛瑞涛,刘宁,杨晓莉,魏从文,钟辉,吴飞翔,陈伟,李自慧,邹岭,叶甲舟,白涛,陈洁,陈健康,王翠..重组pEGFP-C1-CXCL1真核表达载体构建及其对内质网应激下人肝癌细胞HepG2的增殖作用[J].军事医学,2017,41(10):792-795,4.基金项目
广西卫生和计划生育委员会重点课题资助项目(S201513) （S201513）
区域性高发肿瘤早期防治重点实验室子课题资助项目(GKZ201604) （GKZ201604）
广西科学技术厅重点研发课题资助项目(桂科AB16380242) （桂科AB16380242）
