【摘要】 目的:研究腺病毒载体AdING4对人MCF-7乳腺癌细胞的生长抑制及化疗增敏作用。方法:将搭载有ING-4基因的重组腺病毒载体AdING4感染人MCF-7乳腺癌细胞,用荧光显微镜观察感染后的MCF-7细胞形态学变化;RT-PCR和Western-Blot法检测ING-4基因在MCF-7细胞中的转录和表达;RT-PCR法检测凋亡相关基因在MCF-7细胞中的表达;CCK法测定Ad-ING4感染MCF-7乳腺癌细胞后所发挥的细胞增殖抑制作用。流式细胞技术检测ING-4对MCF-7乳腺癌细胞的促凋亡作用。CCK-8法分别测定病毒感染前后的MCF-7乳腺癌细胞的药物半数抑制浓度IC50,并观察Ad-ING4与化疗药物合用后对MCF-7细胞增殖抑制和化疗增敏现象。结果:MCF-7细胞在转染ING-4基因后,明显出现变圆、脱落、皱缩、聚集等现象;外源性ING-4基因在MCF-7细胞中获得成功表达;外源性ING-4基因作用下MCF-7细胞的增殖受到了明显抑制,凋亡率有所升高,凋亡相关基因Bax的表达水平明显上调,Bcl-2、Survivin的表达水平明显下调。ING-4基因感染MCF-7细胞后,使MCF-7细胞对相关化疗药物的敏感度更高;ING-4基因与化疗药物合用后对MCF-7细胞的增殖抑制作用,较之单用化疗药物更为明显。结论:MCF-7细胞在转染ING4基因后其增殖受到了明显抑制并更易凋亡,该现象可能是通过改变Bax,Bcl-2及Survivin表达水平来实现的,且对化疗药物的敏感性更高。更多还原

【Abstract】 Objective:To explore the growth-inhibitory and enhanced chemotherapy sensitization effects of the adenovirus vector AdING4 on the MCF-7 human breast cancer cells of the adenovirus-mediated ING4 gene.Methods:The MCF-7 human breast cancer cells were infected with AdING4 and observed by fluorescence microscope.RT-PCR and Western-Blot analysis were selected to detect the transcription of ING4 gene.The expression levels of apoptosis-related genes were detected by RT-PCR analysis.The cell growth inhibition effects of AdING4 on the MCF-7 cells were detected by CCK-8 assay.Cell apoptosis was detected by flow cytometry assay.CCK-8 assay was used to detect the IC50 of MCF-7 cells infected by ING4 or not.The phenomenon of cell growth inhibition and enhanced chemotherapy sensitization was observed after the AdING4-Drugs combined application.Results:The ING4 gene was successfully transfected into MCF-7 cells,and the cell-growth of MCF-7 cells infected by AdING4 was obviously inhibited,and the infected cells had a higher apoptosis rate.RT-PCR analysis showed that Bax was up-regulated and Bcl-2 and Survivin were down-regulated.The MCF-7 cells infected by AdING4 were more sensitive to chemotherapy drugs than those without infection.The Cell growth inhibition effects were obviously reinforced by AdING4.Conclusion:AdING4 can inhibit the growth of MCF-7 cells and induce cell apoptosis,which may result in changing expression levels of Bax and Bcl-2.And it also resulted in enhancing chemotherapy sensitization effects.更多还原