生物技术通报2018，Vol.34Issue(1)：247-252,6.DOI:10.13560/j.cnki.biotech.bull.1985.2017-0557
SGK3基因RNAi慢病毒载体的构建及其对乳腺癌MB-474细胞增殖和凋亡的影响
Construction of SGK3 Gene Lentiviral RNA Interference Vector and Effects on Cell proliferation and Apoptosis of Breast Cancer Cell Line MB-474
摘要
Abstract
This study is aimed to construct the lentiviral RNA interference(RNAi)vector of SGK3 gene,and to observe its effect on the proliferation and apoptosis of human breast cancer cell line MB-474. The lentiviral vector of shRNA sequence targeting SGK3 was constructed and transfected into human breast cancer MB-474 cells to silence the SGK3 gene. The expression of SGK3 mRNA and protein in transfected MB-474 cells was detected by real-time PCR and Western bloting,and the effect of SGK3 gene silencing on the proliferation,cell cycle and apoptosis of MB-474 cells was detected by MTT assay and flow cytometry. Gene sequencing showed that 4 RNAi lentiviral vectors targeting the SGK3 gene were successfully constructed. The lentiviral vectors were packed successfully by 293T cells and the virus titer was(3-8) ×108 TU/mL. After the recombinant lentiviral vector was transfected into MB-474 cells,real-time PCR and Western bloting demonstrated that the expression of SGK3 in interference group was lower than that of control group. Among the 4 lentiviral vectors,the one with best interference effect is the PGC-LV3-SGK3-1 sequence. SGK3 silencing inhibited MB-474 cell proliferation and promoted apoptosis,but had no significant effect on cell cycle progression. In short,RNAi lentivirus vector targeting SGK3 gene was successfully constructed,and SGK3 gene silencing affected cell proliferation and apoptosis of breast cancer.关键词
RNAi慢病毒/SGK3/细胞增殖/细胞凋亡Key words
RNAi lentivirus/SGK3/cell proliferation/apoptosis引用本文复制引用
郭红艳,高涵,吴琦,孙晓杰,刘秀财,赵立群..SGK3基因RNAi慢病毒载体的构建及其对乳腺癌MB-474细胞增殖和凋亡的影响[J].生物技术通报,2018,34(1):247-252,6.基金项目
黑龙江省教育厅科学技术研究项目(2016-KYYWF-0836) （2016-KYYWF-0836）
