安徽医科大学学报2017，Vol.52Issue(6)：839-842,4.DOI:10.19405/j.cnki.issn1000-1492.2017.06.013
TMEM88真核表达质粒的构建及其功能研究
Construction of TMEM88 eukaryotic expression plasmid and its function research
摘要
Abstract
Objective To construct the eukaryotic expression plasmid TMEM88,and study its effects on cell proliferation and apoptosis of liver cancer cells.Methods Extract total RNA of human hepatic stellate cells,reverse transcription into cDNA as templates,and obtain amplification TMEM88 CDS sequence by PCR,after connect to pEGFP-C2 carrier by using the double enzyme,and then make the conversion,plasmid extraction,and enzyme identification,finally take positive clones to be sequenced in biological company.pEGFP-C2-TMEM88 eukaryotic expression plasmid was transfected respectively to human liver cancer cell line SMMC-7721,then,MTT and flow cytometry methods were used to detect their effects on cell proliferation and apoptosis.Results The sequencing results showed that pEGFP-C2-TMEM88 eukaryotic expression plasmid was built successfully.MTT experiment results showed that the cell proliferation rate of transfection group was(0.625±0.07),was significantly lower than that of normal group (0.880±0.09) (P<0.05);Flow cytometry results showed that cell apoptosis rate of transfection group was 22.1%,significantly higher than that of normal group(9.1%).Conclusion TMEM88 can significantly inhibit the proliferation of human liver cancer cell line SMMC-7721,and promote its apoptosis,further understand the function of TMEM88,and lay a foundation for searching a new direction in the therapy of liver cancer.关键词
TMEM88/转染/增殖/凋亡Key words
TMEM88/transfection/proliferation/apoptosis分类
医药卫生引用本文复制引用
倪升丽,胡富勇,李增,孟晓明,王学富,徐涛..TMEM88真核表达质粒的构建及其功能研究[J].安徽医科大学学报,2017,52(6):839-842,4.基金项目
合肥市科技攻关计划项目(编号:合科[2015]163号-38) （编号:合科[2015]163号-38）
安徽医科大学博士科研资助基金(编号:XJ201536) （编号:XJ201536）
安徽高校自然科学研究重点项目(编号:KJ2016A348) （编号:KJ2016A348）
