miR-660-5p靶向TET2促进乳腺癌的增殖、迁移和侵袭

miR-660-5p Targets TET2 to Promote Breast Cancer Proliferation,Migration and Invasion

目的研究miR-660-5p/TET2通过PI3K/AKT/mTOR信号通路调节乳腺癌细胞增殖、凋亡、迁移和侵袭的机制。方法收集2021年1月-2022年6月荆门市人民医院65例乳腺癌患者的乳腺癌组织和邻近正常组织样品,qPCR实验检测miR-660-5p在乳腺癌组织和细胞系中的表达水平,Kaplan-Meier生存分析和Log-rank检验分析miR-660-5p与患者临床病理特征的相关性;CCK8、流式细胞术和Transwell方法检测miR-660-5p对乳腺癌细胞增殖、凋亡、迁移和侵袭情况;双荧光素酶报告基因和蛋白质印迹实验明确miR-660-5p与TET2的关系;Western blot、CCK8和Transwell实验明确miR-660-5p通过靶向TET2调节乳腺癌的进程;Western blot实验检测miR-660-5p/TET2是否激活PI3K/AKT/mTOR信号通路;动物成瘤实验证明敲低miR-660-5p在乳腺癌中的作用。结果miR-660-5p在乳腺癌组织中表达上调,且miR-660-5p的高表达与乳腺癌的淋巴结转移(P=0.003),晚期TNM分期(P=0.009)和血管浸润(P=0.018)密切相关;敲低miR-660-5p可抑制乳腺癌细胞增殖、迁移和侵袭,诱导乳腺癌细胞凋亡;TET2是miR-660-5p的直接靶标,干扰TET2可以部分逆转敲低miR-660-5p对乳腺癌细胞恶性潜能的抑制作用;miR-660-5p下调TET2并激活PI3K/AKT/mTOR信号通路;敲低miR-660-5p可抑制体内肿瘤生长。结论miR-660-5p通过靶向TET2和PI3K/AKT/mTOR信号通路来促进乳腺癌的进程,可能是治疗乳腺癌的潜在靶标。

Objective To study the mechanism of miR-660-5p/TET2 regulating the proliferation,apoptosis,migration and invasion of breast cancer cells through PI3K/AKT/mTOR signaling pathway.Methods The samples of breast cancer tissues and adjacent normal tissues of 65 patients with breast cancer in Jingmen People's Hospital from January 2021 to June 2022 were collected.The expression level of miR-660-5p in breast cancer tissues and cell lines was detected by qPCR.Kaplan-Meier survival analysis and log-rank test were used to analyze the correlation between miR-660-5p and clinicopathological features of patients.The effects of miR-660-5p on the proliferation,apoptosis,migration and invasion of breast cancer cells were detected by CCK8,flow cytometry and Transwell methods.The relationship between miR-660-5p and TET2 was determined by dual luciferase reporter gene and Western blotting.Western blot,CCK8 and Transwell experiments were used to confirm that miR-660-5p regulated breast cancer progression by targeting TET2.Whether miR-660-5p/TET2 activated PI3K/AKT/mTOR signal pathway was detected by Western blot experiment.The effect of knockdown of miR-660-5p in breast cancer was further proved through animal tumor formation experiment.Results The expression of miR-660-5p was up-regulated in breast cancer tissues,and the high expression of miR-660-5p was closely related to the lymph node metastasis(P=0.003),advanced TNM staging(P=0.009)and vascular invasion(P=0.018)of breast cancer.Knockdown of miR-660-5p inhibited breast cancer cell proliferation,migration and invasion,and induced breast cancer cell apoptosis.TET2 was the direct target of miR-660-5p,interference with TET2 could partially reverse the inhibitory effect of knockdown miR-660-5p on the malignant potential of breast cancer cells.miR-660-5p down-regulated TET2 and activated PI3K/AKT/mTOR signaling pathway.Knockdown of miR-660-5p inhibited tumor growth in vivo.Conclusions miR-660-5p promotes breast cancer progression by targeting TET2 and PI3K/AKT/mTOR signaling pathways,and may be a potential target for breast cancer treatment.