GP73通过p53信号通路调节乳腺癌细胞MCF-7的增殖和凋亡

GP73 regulates proliferation and apoptosis of breast cancer MCF-7 via p53 signaling pathway

目的探讨高尔基蛋白73(GP73)对乳腺癌细胞MCF-7的增殖、凋亡的影响及相关机制。方法利用干扰及过表达GP73质粒转染乳腺癌细胞MCF-7,并利用CCK-8检测处理后细胞的增殖能力,利用流式细胞术方法检测处理组细胞的凋亡水平,利用qRT-PCR和Western印迹法测定GP73和p53 mRNA和蛋白的表达。采用qRT-PCR测定20例临床乳腺癌和癌旁组织中GP73和p53的mRNA水平,并分析乳腺癌组织中GP73和p53的mRNA水平的相关性。结果过表达GP73后MCF-7较对照组增殖能力上升(P<0.05),凋亡水平下降(P<0.05);干扰GP73后MCF-7增殖能力下降(P<0.05),凋亡率升高。同时过表达GP73和p53的乳腺癌细胞较单独过表达GP73的MCF-7细胞的增殖能力下降,凋亡水平上升。乳腺癌组织中GP73和p53 mRNA表达也呈负相关(r=-0.528,P<0.01)。结论 GP73可能通过p53信号通路调节乳腺癌细胞MCF-7的增殖和凋亡。

Objective To determine the impact of Golgi protein 73 （GP73） on the proliferation and apoptosis of breast cancer MCF-7 cells and investigate its related mechanism. Methods After MCF-7 cells were separately transfected with GP73 shRNA and overexpression plasmids, CCK-8 assay and flow cytometry were employed to detect the proliferation and apoptosis, respectively in the transfected cells. The expression of GP73 and p53 at mRNA and protein levels was determined by qRT-PCR and Western blotting. The mRNA expression of above 2 molecules was detected in 20 breast cancer tissues and paired paracancerous tissues with qRT-PCR. The correlation of GP73 and p53 mRNA levels in breast cancer tissue were analyzed. Results GP73 overexpression enhanced the proliferation and suppressed the apoptosis of MCF-7 cells, while GP73 down-regulation inhibited the proliferation and improved apoptosis of MCF-7 cells. Further, co-overexpression of GP73 and p53 resulted in reduced proliferation and increased apoptosis when compared with the MCF-7 cells overexpressed with GP7 or p53 alone. A negative correlation was observed between the mRNA expression levels of GP73 and p53 in breast cancer tissues. Conclusion GP73 may regulate proliferation and apoptosis of breast cancer MCF-7 cells via p53 signaling pathway.