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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。

英文摘要:

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 Ad ING4 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.

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