YAP 基因干扰对乳腺癌细胞增殖和凋亡的影响

YAP gene interference on proliferation and apoptosis in breast cancer cells

目的:探讨干扰YAP基因的表达对乳腺癌细胞增殖和凋亡的影响。方法:通过逆转录病毒介导的方法,分别用YAP shRNA(实验组)或阴性对照shRNA(对照组)转染MCF-7乳腺癌细胞,用qRT-PCR和Western blot方法检测干扰效率;溴脱氧核苷尿嘧啶(BrdU)结合实验和MTT法检测细胞增殖活性;流式细胞术和DAPI染色检测细胞凋亡情况。结果:干扰效率检测显示,转染72 h后,实验组MCF-7细胞YAP mRNA表达量明显降低(P<0.05),同时YAP蛋白表达也明显下调;细胞增殖检测显示,与对照组比较,实验组MCF-7细胞BrdU结合与OD值明显降低(P<0.05),转染后24、48、72 h增殖抑制率分别为19.1%、38.5%、53.5%;凋亡检测显示,实验组细胞凋亡率与凋亡细胞数较对照组明显增加(均P<0.05)。结论:干扰YAP基因的表达能有效抑制乳腺癌细胞的增殖并促进凋亡,YAP可能是乳腺癌潜在治疗靶点。

Objective： To observe the influence of interference of Yes-associated protein （YAP） gene expression on proliferation and apoptosis in breast cancer cells.Methods： Using a retrovirus-mediated method, breast cancer MCF-7 cells were transfected with YAP shRNA （experimental group） or negative control shRNA （control group）, respectively. After that, the interference efficacy was evaluated via qRT-PCR and Western blot analysis, cell proliferative ability was determined by bromodeoxyuridine （BrdU） incorporation assay and MTT assay, and cell apoptosis was detected with flow cytometry and DAPI staining, respectively.Results： Interference efficacy determination showed that YAP mRNA expression level was significantly decreased （P〈0.05）, and YAP protein expression was also remarkably down-regulated in MCF-7 cells in experimental group compared with control group at 72 h after transfection. Cell proliferation assay showed that BrdU incorporation level and OD value in experimental group were significantly reduced compared with control group （both P〈0.05）, and the ratio of proliferation inhibition in experimental group at 24, 48, 72 h after transfection was 19.1%, 38.5% and 53.5%, respectively. Cell apoptosis analysis showed that the apoptosis rate and number of apoptotic cells in experimental group were significantly increased compared with control group （both P〈0.05）.Conclusion： Interference of YAP gene expression can effectively inhibit the proliferation and accelerate apoptosis in breast cancer cells, and YAP may probably a potential therapeutic target for breast cancer.