下调YAP1基因对子宫内膜癌顺铂耐药细胞的影响及机制探究  

Effect of down-regulation of YAP1 gene on cisplatin-resistant endometrial cancer cells and its mechanism

作　　者：谢承宏 陈爱华[1] 于江宇 XIE Chenghong;CHEN Aihua;YU Jiangyu(The People's Hospital of China Three Gorges University·The First People's Hospital of Yichang,Yichang,Hubei443000,China)

机构地区：[1]三峡大学人民医院·宜昌市第一人民医院,湖北宜昌443000

出　　处：《中国优生与遗传杂志》2023年第1期15-21,共7页Chinese Journal of Birth Health & Heredity

摘　　要：目的观察下调YAP1基因对子宫内膜癌顺铂耐药细胞的影响。方法子宫内膜癌顺铂耐药细胞株Ishikawa/DDP分为Ishikawa/DDP-siRNA-YAP1组(转染siRNA-YAP1的细胞)、阴性对照组(转染siRNA-NC的细胞)和空白对照组(未经转染的细胞),MTT法及癌细胞单克隆形成数目检测细胞增殖,Transwell小室检测侵袭,划痕检测迁移,AnnexinV-FITC/PI双染法检测凋亡,免疫荧光检测自噬现象,Western blot检测凋亡相关蛋白Bax、Bcl-2和自噬相关蛋白LC3β、p62。结果24 h、48 h、72 h,Ishikawa/DDP-siRNA-YAP1组OD值明显低于阴性对照组和空白对照组(P<0.05),细胞克隆数明显少于阴性对照组和空白对照组(P<0.05);细胞迁移率明显低于阴性对照组和空白对照组(P<0.05);细胞侵袭数明显少于阴性对照组和空白对照组(P<0.05);免疫荧光结果显示与阴性对照组和空白对照组比较,Ishikawa/DDP-siRNA-YAP1组绿色、红色以及黄色光点明显减少,即自噬小体堆积明显减少,Ishikawa/DDP-siRNA-YAP1组自噬蛋白LC3β和p62蛋白表达明显低于阴性对照组和空白对照组(P<0.05);细胞凋亡率高于阴性对照组和空白对照组(P<0.05),凋亡蛋白Bcl-2表达明显低于阴性对照组和空白对照组,Bax表达明显高于阴性对照组和空白对照组(P<0.05)。结论下调YAP1可能通过抑制自噬,从而抑制细胞增殖、侵袭以及迁移,同时促进凋亡,YAP1有可能成为逆转子宫内膜癌化疗耐药的新靶点。Objective To observe the effect of down-regulating YAP1 gene on cisplatin resistance of endometrial cancer cells by regulating autophagy.Methods Endometrial cancer cisplatin-resistant cell line Ishikawa/DDP was divided into Ishikawa/DDP-siRNA-YAP1 group(cells transfected with siRNA-YAP1),negative control group(cells transfected with siRNA-NC)and blank control group(cells not transfected).Cell proliferation was detected by MTT assay and the number of cancer cell clones,invasion was detected by Transwell chamber,and migration was detected by scratch.AnnexinV-FITC/PI double staining was used to detect apoptosis,immunofluorescence was used to detect autophagy,and Western blot was used to detect apoptosis-related proteins Bax and Bcl-2 and autophagy-related proteins LC3β and p62.Results At 24 h,48 h and 72 h,the OD value of Ishikawa/DDP-siRNA-YAP1 group was significantly lower than that of negative control group and blank control group(P<0.05),and the number of cell clones was significantly less than that of negative control group and blank control group(P<0.05).Cell mobility was significantly lower than that of negative control group and blank control group(P<0.05).The number of cell invasion was significantly less than that of negative control group and blank control group(P<0.05).Compared with the negative control group and the blank control group,the fluorescence results showed that the green,red and yellow spots in Ishikawa/DDP-siRNA-YAP1 group were significantly reduced,that is,the accumulation of autophagy bodies was significantly reduced,and the expressions of LC3β and p62 proteins in Ishikawa/DDP-siRNA-YAP1 group were significantly lower than those in the negative control group and the blank control group(P<0.05).The apoptosis rate was higher than that of the negative control group and the blank control group(P<0.05),the expression of apoptosis protein Bcl-2 was significantly lower than that of the negative control group and the blank control group,and the expression of Bax was significantly higher than tha

关 键 词：YAP1基因 自噬 子宫内膜癌细胞顺铂耐药 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]