miR-424-3p靶向SMAD7调控子宫内膜癌增殖、侵袭机制研究

Study on mechanism of miR-424-3p targeting SMAD7 in regulating proliferation and invasion of endometrial carcinoma

摘要

目的 探讨miR-424-3p是否通过介导SMAD7调控子宫内膜癌(EC)的增殖、侵袭,并探究调控机制。方法 利用TargetScan数据和GEPIA数据库分别进行预测性结合位点分析和基因表达评估。通过EdU染色、伤口愈合和Transwell试验评估细胞的增殖、迁移和侵袭。通过移植肿瘤实验在体内评价miR-424-3p和SMAD对EC的调控作用。结果 我们证实了miR-424-3p与SMAD7的结合位点。敲除miR-424-3p能显著抑制HEC-1A和RL95-2细胞的侵袭和增殖,同时促进细胞凋亡。然而,sh-SMAD7逆转了miR-424-3p对这些细胞的影响。此外,sh-SMAD7还能显著抵消miR-424-3p抑制剂对HEC-1A和RL95-2细胞系上皮-间质转化(EMT)相关蛋白表达的影响。miR-424-3p抑制剂能够显著抑制移植瘤的生长,但sh-SMAD7可逆转miR-424-3p抑制剂的影像。结论 miR-424-3p通过调控SMAD7影响HEC-1A和RL95-2细胞的增殖、侵袭和上皮间质转化过程,提示miR-424-3p/SMAD7可能是子宫内膜癌的防治的新靶点。

Abstract

Objective To investigate whether miR-424-3p regulates the proliferation and invasion of endometrial carcinoma (EC) by mediating SMAD7,and to explore the regulatory mechanism. Methods Predictive binding site analysis and gene expression assessment were conducted using TargetScan data and the GEPIA database. Cell proliferation,migration,and invasion were evaluated through EdU staining,wound healing,and Transwell assays. The regulatory effects of miR-424-3p and SMAD on EC were evaluated in vivo through tumor transplantation experiments. Results Validation confirmed the binding site between miR-424-3p and SMAD7. Knockout of miR-424-3p significantly inhibited the invasion and proliferation of HEC-1A and RL95-2 cells,while promoting cell apoptosis. However,sh-SMAD7 reversed the effects of miR-424-3p on these cells. Additionally,sh-SMAD7 could significantly counteract the effects of miR-424-3p inhibitors on the expression of epithelial-mesenchymal transition (EMT)-related proteins in HEC-1A and RL95-2 cell lines. miR-424-3p inhibitors could significantly inhibit the growth of transplanted tumors,but sh-SMAD7 could reverse the effects of miR-424-3p inhibitors. Conclusion miR-424-3p regulates the proliferation,invasion,and epithelial-mesenchymal transition process of HEC-1A and RL95-2 cells by modulating SMAD7,suggesting that miR-424-3p/SMAD7 may be a new target for the prevention and treatment of endometrial carcinoma.

关键词

miR-424-3p / 子宫内膜癌 / SMAD7 / 上皮间质转化 / 增殖 / 侵袭

Key words

miR-424-3p / endometrial carcinoma / SMAD7 / epithelial-mesenchymal transition / proliferation / invasion

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基金资助

中国人民解放军第22医院自主科研基金项目()