METHODS: The expression levels of miR-34b-5p and zinc finger E-box-binding homeobox 1 (ZEB1) in endometrial cancer cells were analyzed by qRT-PCR, and ZEB1 expression in endometrial cancer tissues was examined by immunohistochemistry. Proliferation, migration, and invasion of endometrial cancer AN3CA cells were evaluated using CCK8, scratch, and transwell assays, respectively. Bioinformatic analysis and dual-luciferase reporter gene assays were used to validate the targeting relationship between miR-34b-5p and ZEB1. Western blotting was performed to analyze the expression levels of ZEB1 and EMT-related proteins.
RESULTS: miR-34b-5p was significantly downregulated in endometrial cancer AN3CA cells. Overexpression of miR-34b-5p significantly inhibited proliferation, invasion, migration, and the EMT of endometrial cancer AN3CA cells. ZEB1, which was identified as a direct target gene of miR-34b-5p, exhibited high expression in endometrial cancer cells and tissues. Additionally, ZEB1 upregulation partially reversed the inhibitory effects of miR-34b-5p on proliferation, migration, invasion, and the EMT of endometrial cancer AN3CA cells.
CONCLUSIONS: miR-34b-5p suppresses the EMT and metastasis in endometrial cancer AN3CA cells by targeting ZEB1, indicating that the miR-34b-5p-ZEB1-EMT axis may be a therapeutic target for endometrial cancer.
方法:通过qRT-PCR分析子宫内膜癌细胞中miR-34b-5p和锌指E盒结合同源盒1(ZEB1)的表达水平,和ZEB1在子宫内膜癌组织中的表达通过免疫组织化学检查。扩散,迁移,用CCK8、划痕、和transwell分析,分别。生物信息学分析和双荧光素酶报告基因检测用于验证miR-34b-5p和ZEB1之间的靶向关系。进行蛋白质印迹以分析ZEB1和EMT相关蛋白的表达水平。
结果:miR-34b-5p在子宫内膜癌AN3CA细胞中显著下调。miR-34b-5p过表达显著抑制细胞增殖,入侵,迁移,和子宫内膜癌AN3CA细胞的EMT。ZEB1被鉴定为miR-34b-5p的直接靶基因,在子宫内膜癌细胞和组织中呈高表达。此外,ZEB1上调部分逆转了miR-34b-5p对增殖的抑制作用,迁移,入侵,和子宫内膜癌AN3CA细胞的EMT。
结论:miR-34b-5p通过靶向ZEB1抑制子宫内膜癌AN3CA细胞的EMT和转移,提示miR-34b-5p-ZEB1-EMT轴可能是子宫内膜癌的治疗靶点。