中国肿瘤生物治疗杂志2017，Vol.24Issue(6)：608-614,7.DOI:10.3872/j.issn.1007-385X.2017.06.006
稳定过表达人MGST1基因抑制肺腺癌细胞SPC-A-1的凋亡
Stable overexpression of human MGST1 gene inhibits apoptosis of lung adenocarcinoma cell line SPC-A-1
摘要
Abstract
Objective:Lung adenocarcinoma SPC-A-1 cell line that stably over-expresses microsomal glutathione Stransferase 1 (MGST1) was constructed to explore the function and mechanism of MGST1 in lung adenocarcinoma.Methods:The recombinant plasmid pcDNA3-MGST1 and the empty vector pcDNA3 were transfected into SPC-A-1 by Lipofectamine-mediated method;after being screened by G418,stably transfected cells were labeled and divided into pcDNA3-MGST1 group and empty vector pcDNA3 group.qRT-PCR and Western blotting were used to detect the expression of MGST1 at mRNA and protein level in stable cells.MTS was used to detect the viability of cells.Flow cytometry and Western blotting were used to detect the cell apoptotic rate and the downstream apoptotic proteins induced by H2O2,respectively.Results:The results of sequencing showed that the recombinant plasmid pcDNA3-MGST1 was successfully constructed,and the stable cell line with G418 resistance was obtained.The mRNA and protein level of MGST1 in pcDNA3-MGST1 group were all significantly elevated (P ＜0.01),and the cell viability was significantly increased (P ＜ 0.05).Under the induction of H2O2,the early apoptosis rate of pcDNA3-MGST1 group was significantly lower than that of pcDNA3 group ([3.30 ± 0.40] ％ vs [6.50 ± 0.95] ％,P ＜ 0.05).The expressions of apoptotic proteins (caspase 9,caspase3 and PARP) increased,and the expressions of cleaved-caspase 9,cleaved-caspase 3 and cleaved-PARP were significantly decreased in pcDNA3-MGST1 group.Conclusion:The lung adenocarcinoma cell line SPC-A-1,which stably over-expresses human MGST1,has been successfully established.It was found that MGST1 could inhibit the apoptosis of lung adenocarcinoma cells by regulating caspase apoptosis pathway,which laid the experimental foundation for the followup study.关键词
肺腺癌/微粒体谷胱甘肽S转移酶1/基因克隆/稳定过表达/caspase蛋白/凋亡Key words
lung adenocarcinoma/microsomal glutathione S-transferase 1 (MGST1)/gene clone/stable overexpression/caspase protein/apoptosis分类
医药卫生引用本文复制引用
曾宝真,葛春蕾,付桥粉,张志伟,宋鑫,黄文华..稳定过表达人MGST1基因抑制肺腺癌细胞SPC-A-1的凋亡[J].中国肿瘤生物治疗杂志,2017,24(6):608-614,7.基金项目
国家自然科学基金资助项目(No.U1502222,No.81470005,No.81260307) （No.U1502222,No.81470005,No.81260307）
国家自然科学基金重大科研仪器研制项目(No.61427807).Project supported by the National Natural Science Foundation of China(No.U1502222,No.81470005,No.81260307),and the key Scientific Research Equipment Development Project of the National Natural Science Foundation of China(No.61427807) （No.61427807）
