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 that of the negative control group and the blank control group(P<0.05).Conclusion Down-regulation of YAP1may inhibit cell proliferation,invasion and migration by inhibiting autophagy,and promote apoptosis at the same time.YAP1may become a new target for reversing chemotherapy resistance of endometrial cancer cells.
作者
谢承宏
陈爱华
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)
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
关键词
YAP1基因
自噬
子宫内膜癌细胞顺铂耐药
凋亡
YAP1 gene
autophagy
cisplatin resistance in endometrial cancer cells
apoptosis
