GSTM3过表达或沉默的乳腺癌细胞株MDA-MB-231的构建  

Overexpressing and Silencing of GSTM3 in MDA-MB-231 Breast Cancer Cell Line

作　　者：黄秋梅[1] 徐晓晶[2] 阳国平[3] 赵莹[2] 周玲[2] 王嘉颖[2] 袁洪[3] 金维荣[2] 

机构地区：[1]中南大学药学院药理学系,长沙410078 [2]生物芯片上海国家工程研究中心,上海201203 [3]中南大学湘雅三医院,长沙410013

出　　处：《复旦学报（自然科学版）》2009年第5期674-679,共6页Journal of Fudan University：Natural Science

基　　金：国家自然科学基金资助项目(30800291);上海市青年科技启明星计划资助项目(07QB14015)

摘　　要：为深入研究谷胱甘肽S-转移酶μ3(glutathione S-transferase M3,GSTM3)在乳腺癌中的作用,构建了真核重组表达质粒pcDNA3.1/GSTM3,并利用真核表达载体pSilencer2.1U6-neo构建GSTM3特异性siRNA表达载体.重组质粒转染MDA-MB-231细胞,半定量、荧光定量RT-PCR和Western blot对转染细胞的检测结果表明,转染GSTM3基因真核表达质粒的细胞株中GSTM3表达明显提高,而转染siRNA表达质粒后,GSTM3的表达则受到显著抑制.To investigate the biological roles of GSTM3 on breast cancer carcinogenesis,MDA-MB-231 breast cell lines either GSTM3-stably overexpressed or silenced were established.GSTM3 gene was subcloned into pcDNA3.1 mammalian expression vector to obtain recombinant pcDNA3.1/GSTM3.The oligonucleotide sequences encoding the GSTM3-specific shRNA were designed and subcloned into pSilencer2.1-U6 neo vector.MDA-MB-231 cell lines were transfected with the recombinant vectors.Expression level of GSTM3 mRNA and protein were analyzed by semi-quantitative RT-PCR,real time RT-PCR and Western blot.Overexpression of GSTM3 was detectable in cell lines transfected with pcDNA3.1/GSTM3,whereas GSTM3 expression was significantly decreased in the siRNA interfered cell lines.

关 键 词：谷胱甘肽S-转移酶μ3 乳腺癌 RNA干扰 

分 类 号：Q969.436.5[生物学—昆虫学] S858.292[农业科学—临床兽医学]