摘要
目的 以逆转录病毒为载体,研究抑瘤素M(oncostatin M,OSM)基因对人乳腺癌细胞株(michigan cancer foundation-7,MCF-7)生长抑制作用.方法 以最佳感染剂量携带OSM基因的逆转录病毒感染MCF-7人乳腺癌细胞,反转录酶-聚合酶链锁反应(reverse transcription-polymerase chain reaction,RT-PCR)和酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测目的基因在MCF-7人乳腺癌细胞中的转录;对感染后的MCF-7人乳腺癌细胞,以四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium,MTT)检测OSM基因对肿瘤细胞的生长抑制作用,流式法检测OSM基因诱导肿瘤细胞的凋亡效应,赫斯特(Hochest)染色法检测凋亡细胞的核形态改变,并用RT-PCR法进一步检测肿瘤细胞中凋亡相关基因的表达.结果 逆转录病毒载体能介导OSM基因在MCF-7人乳腺癌细胞中的转录及表达后,能够显著的抑制肿瘤细胞的生长(MTT法,P=0.0162,P<0.05)并诱导其凋亡(膜联蛋白/碘化丙啶双染法,P=0.02,P<0.05).结论 OSM基因对MCF-7细胞有显著的抑制作用,其作用的机制可能与其上调凋亡促进蛋白Bax和Caspase3,下调凋亡调控基因Bcl-2有关.
Objective To study the inhibition effects of oncostatin M (OSM) gene on human breast cancer cell line(michigan cancer foundation-7,MCF-7).Methods MCF-7 cells were infected with the optimized dose of retrovirus expressing OSM.Expression of the target gene was determined by reverse transcriptionpolymerase chain reaction(RT-PCR) and enzyme-linked immunosorbent assay(ELISA).Methyl thiazolyl tetrazolium (MTT) assay was employed to examine growth inhibition effect of OSM gene on MCF-7 cells.Cell apoptosis was determined by flow cytometry.The morphologic changes of nucleus was detected by Hochest staining,in addition,the expression of apoptosis related genes in tumor cells were further detected by RT-PCR.Results OSM gene was well expressed in human breast cancer MCF-7 cell by retrovial vetor and it could significantly inhibit the growth of MCF-7 cell(MTT,P =0.0162,P < 0.05) and induce its apoptosis (Annexin V/PI,P =0.02,P < 0.05).Conclusion OSM gene has significant inhibitory effects on MCF-7 cells,which mechanism might be related to the upregulation of Bax and Caspase3 and the downregulation of Bcl-2.
出处
《中华内分泌外科杂志》
CAS
2013年第5期388-391,共4页
Chinese Journal of Endocrine Surgery