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目的检测ING4(inhibitor of growth 4)对MCF-7细胞凋亡的影响。方法从人胎盘总RNA中克隆人ING4基因,构建其真核表达质粒pEGFP-ING4转染MCF-7细胞表达ING4后,流式细胞仪检测细胞凋亡率;荧光定量RT-PCR检测ING4、NF-κB、caspase-3、IL-8、Bcl-2及Bax基因的表达。结果构建的重组质粒转染可见目的蛋白融合表达,与对照相比MCF-7细胞凋亡率增高;ING4、caspase-3及IL-8基因的表达上调,NF-κB与Bcl-2/Bax基因的表达下调。结论成功从人胎盘组织克隆了ING4基因并构建其真核表达质粒在人MCF-7细胞中表达;ING4在人MCF-7细胞中能引起凋亡相关基因的改变并促进MCF-7细胞的凋亡,这为进一步研究ING4的抗肿瘤机制奠定了基础。
Objective To investigate the effect of inhibitor of growth 4 on apoptosis of MCF-7 cells. METHODS: The human ING4 gene was cloned from human placental total RNA. The eukaryotic expression plasmid pEGFP-ING4 was transfected into MCF-7 cells to express ING4. Flow cytometry was used to detect the apoptosis rate. ING4 and NF -κB, caspase-3, IL-8, Bcl-2 and Bax gene expression. Results The recombinant plasmids were transfected with the target protein. The apoptosis rate of MCF-7 cells was increased compared with that of the control. The expression of ING4, caspase-3 and IL-8 genes was up-regulated. The expressions of NF-κB and Bcl- Down-regulated CONCLUSION: ING4 gene was successfully cloned from human placenta tissue and its eukaryotic expression plasmid was constructed and expressed in human MCF-7 cells. ING4 can induce apoptosis-related genes and promote apoptosis of MCF-7 cells in human MCF-7 cells This laid the foundation for further study of anti-tumor mechanism of ING4.