YAP1靶向抑制剂CA3对人子宫内膜癌细胞增殖和凋亡的影响及分子机制

Effects and Molecular Mechanism of YAP1 Targeting Inhibitor CA3 on Proliferation and Apoptosis of Human Endometrial Cancer Cell Lines

关键词:  CA3  Ishikawa细胞  RL95-2细胞  细胞增殖  细胞凋亡

Key words:  CA3  Ishikawa cell  RL95-2 cell  cell proliferation  cell apoptosis

摘要:

目的 探讨YAP1靶向抑制剂CA3对人子宫内膜癌Ishikawa和RL95-2细胞增殖和凋亡的影响及分子机制。方法 采用不同浓度CA3处理Ishikawa和RL95-2细胞,利用CCK-8和克隆形成试验检测细胞增殖能力,微板检测系统检测细胞内ROS水平,流式细胞术检测细胞凋亡,Western blotting检测YAP1、MCM5、Bax和Bcl-2蛋白表达水平。结果 与对照组相比,CA3处理组能够明显降低细胞生存率、减少克隆形成数目、增强细胞内ROS水平和诱导细胞凋亡,下调YAP1、MCM5和Bcl-2蛋白表达水平,但对Bax蛋白表达水平没有影响。结论 CA3靶向沉默YAP1抑制细胞增殖和诱导细胞凋亡,起到明显的抗肿瘤作用,有望成为临床干预和治疗子宫内膜癌的潜在药物。

Abstract:

OBJECTIVE To investigate the effects and molecular mechanism of YAP1 targeting inhibitor CA3 on proliferation and apoptosis of human endometrial cancer cell lines Ishikawa and RL95-2. METHODS Ishikawa and RL95-2 cells were treated with different doses of CA3, and cell proliferation was determined by cell counting kit-8 assay and colony formation assay. The level of intracellular ROS was detected using micro-plate detecting system and cell apoptosis was analyzed by flow cytometry. The expression levels of YAP1, MCM5, Bax and Bcl-2 protein were tested using Western blotting. RESULTS Compared with control group, CA3 treatment groups could significantly inhibit cell survival rate and the number of clone formation, increase the level of intracellular ROS, induce cell apoptosis, down-regulate the expression levels of YAP1, MCM5 and Bcl-2 protein, but the expression of Bax protein did not be affected. CONCLUSION CA3 targets silencing of YAP1 inhibits cell proliferation and induces cell apoptosis, exerts significant anticancer effects, which may be potential drug for clinical prevention and clinic therapy of human endometrial carcinoma.