中华内分泌外科杂志
中華內分泌外科雜誌
중화내분비외과잡지
CHINESE JOURNAL OF ENDOCRINE SURGERY
2013年
5期
388-391
,共4页
焦良和%刘庆红%朱正才%乔纯忠
焦良和%劉慶紅%硃正纔%喬純忠
초량화%류경홍%주정재%교순충
逆转录病毒载体%抑瘤素M%乳腺癌%细胞增殖%凋亡
逆轉錄病毒載體%抑瘤素M%乳腺癌%細胞增殖%凋亡
역전록병독재체%억류소M%유선암%세포증식%조망
Retroviral vector%OSM%Breast cancer%Proliferation%Apoptosis
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.
目的 以逆轉錄病毒為載體,研究抑瘤素M(oncostatin M,OSM)基因對人乳腺癌細胞株(michigan cancer foundation-7,MCF-7)生長抑製作用.方法 以最佳感染劑量攜帶OSM基因的逆轉錄病毒感染MCF-7人乳腺癌細胞,反轉錄酶-聚閤酶鏈鎖反應(reverse transcription-polymerase chain reaction,RT-PCR)和酶聯免疫吸附測定法(enzyme-linked immunosorbent assay,ELISA)檢測目的基因在MCF-7人乳腺癌細胞中的轉錄；對感染後的MCF-7人乳腺癌細胞,以四甲基偶氮唑鹽比色法(methyl thiazolyl tetrazolium,MTT)檢測OSM基因對腫瘤細胞的生長抑製作用,流式法檢測OSM基因誘導腫瘤細胞的凋亡效應,赫斯特(Hochest)染色法檢測凋亡細胞的覈形態改變,併用RT-PCR法進一步檢測腫瘤細胞中凋亡相關基因的錶達.結果 逆轉錄病毒載體能介導OSM基因在MCF-7人乳腺癌細胞中的轉錄及錶達後,能夠顯著的抑製腫瘤細胞的生長(MTT法,P=0.0162,P＜0.05)併誘導其凋亡(膜聯蛋白/碘化丙啶雙染法,P=0.02,P＜0.05).結論 OSM基因對MCF-7細胞有顯著的抑製作用,其作用的機製可能與其上調凋亡促進蛋白Bax和Caspase3,下調凋亡調控基因Bcl-2有關.
목적 이역전록병독위재체,연구억류소M(oncostatin M,OSM)기인대인유선암세포주(michigan cancer foundation-7,MCF-7)생장억제작용.방법 이최가감염제량휴대OSM기인적역전록병독감염MCF-7인유선암세포,반전록매-취합매련쇄반응(reverse transcription-polymerase chain reaction,RT-PCR)화매련면역흡부측정법(enzyme-linked immunosorbent assay,ELISA)검측목적기인재MCF-7인유선암세포중적전록；대감염후적MCF-7인유선암세포,이사갑기우담서염비색법(methyl thiazolyl tetrazolium,MTT)검측OSM기인대종류세포적생장억제작용,류식법검측OSM기인유도종류세포적조망효응,혁사특(Hochest)염색법검측조망세포적핵형태개변,병용RT-PCR법진일보검측종류세포중조망상관기인적표체.결과 역전록병독재체능개도OSM기인재MCF-7인유선암세포중적전록급표체후,능구현저적억제종류세포적생장(MTT법,P=0.0162,P＜0.05)병유도기조망(막련단백/전화병정쌍염법,P=0.02,P＜0.05).결론 OSM기인대MCF-7세포유현저적억제작용,기작용적궤제가능여기상조조망촉진단백Bax화Caspase3,하조조망조공기인Bcl-2유관.
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.
