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目的探究过表达胰岛素样生长因子结合蛋白7(IGFBP7)对人乳腺癌细胞系-7(MCF-7)增殖的影响及其机制。方法采用LipofectamineTM2000将p IRES2-Zs Green1-IGFBP7质粒或p IRES2-Zs Green1空白质粒转染进MCF-7乳腺癌细胞,并用荧光显微镜鉴定细胞转染;实时荧光定量聚合酶链反应(Real-time PCR)对转染48 h后IGFBP7蛋白进行定量;细胞凋亡实验检测转染24、48和72 h后各组细胞增殖情况;Western bolt检测转染48 h后各组细胞(蛋白激酶B/雷帕霉素靶蛋白)(AKT/m TOR)信号通路相关蛋白的表达。结果 Real-time PCR的检测结果提示经过IGFBP7转染的细胞IGFBP7 m RNA表达水平明显升高;细胞凋亡实验结果发现,与空白处理组和空白质粒转染组比较,IGFBP7过表达组细胞增殖能力明显降低;Western bolt检测结果发现,AKT蛋白的磷酸化水平明显下降,且其下游的m TOR表达明显下降(P<0.05)。结论IGFBP7过表达对MCF-7乳腺癌细胞的增殖具有抑制效果,可能是通过对AKT/m TOR信号通路的抑制达到对MCF-7的抑制。
Objective To investigate the effect of overexpression of insulin-like growth factor binding protein 7 (IGFBP7) on proliferation of human breast cancer cell line-7 (MCF-7) and its mechanism. Methods pIRES2-Zs Green1-IGFBP7 plasmid or p IRES2-Zs Green1 blank plasmid was transfected into MCF-7 breast cancer cells by LipofectamineTM2000, and transfection was identified by fluorescence microscopy; Real-time PCR was performed. ) The IGFBP7 protein was quantified after 48 hours of transfection; cell proliferation was detected by apoptosis assay in each group at 24, 48 and 72 hours after transfection; Western blot was used to detect the cells in each group (protein kinase B/Rapa) 48 h after transfection. Expression of AKT/m TOR Signaling Pathway Related Proteins. Results The results of Real-time PCR showed that the expression level of IGFBP7 mRNA was significantly increased in IGFBP7-transfected cells. The results of apoptosis test showed that compared with the blank control group and the blank plasmid transfection group, the IGFBP7 overexpression group had the cell proliferation ability. Significantly reduced; Western bolt detection results showed that the phosphorylation level of AKT protein was significantly decreased, and its downstream m TOR expression was significantly decreased (P <0.05). Conclusion Over-expression of IGFBP7 may have an inhibitory effect on the proliferation of MCF-7 breast cancer cells. It may be through the inhibition of AKT/m TOR signaling pathway to inhibit MCF-7.