微小RNA-145-5p靶向抑制同源形成素样蛋白2基因介导Notch信号通路对口腔鳞状细胞癌细胞增殖、凋亡的调控机制

Study on Mechanism of the Targeted Inhibition of the FMNL2 Mediated Notch Signaling Pathway by MicroRNA-145-5p in Regulating the Proliferation and Apoptosis of Oral Squamous Cell Carcinoma Cells

目的探究微小RNA(miR)-145-5p靶向调控同源形成素样蛋白2(FMNL2)基因介导Notch信号通路对口腔鳞状细胞癌(OSCC)细胞增殖、凋亡的调控机制。方法首先验证miR-145-5p与FMNL2基因之间的靶向调控关系。实时定量聚合酶链反应(qRT-PCR)和蛋白质印迹技术(Western blot)检测癌组织和癌旁组织中miR-145-5p、FMNL2以及Notch通路相关因子(Notch1、Hes1)的表达,干预OSCC细胞中miR-145-5p以及FMNL2基因的表达并对细胞分组。qRT-PCR以及Western blot检测各组细胞中miR-145-5p、FMNL2、Notch通路相关因子以及凋亡相关因子(Bax、Bcl-2)的表达。四甲基偶氮唑盐(MTT)法检测细胞增殖活力。侵袭跨膜(Transwell)实验检测各组细胞侵袭情况。流式细胞术检测各组细胞凋亡。结果FMNL2是miR-145-5p的靶基因。相对于癌旁组织,癌组织中miR-145-5p表达降低,FMNL2基因表达升高,Notch信号通路被激活(P<0.05)。过表达miR-145-5p或沉默FMNL2能够抑制Notch信号通路相关因子的表达,促进Bax增高,Bcl-2降低,同时抑制OSCC细胞增殖和侵袭,促进细胞凋亡(P<0.05)。miR-145-5p抑制组表达与上述变化相反。结论miR-145-5p能够靶向抑制FMNL2基因,进而抑制Notch信号通路的激活,抑制OSCC细胞的增殖、侵袭,促进细胞凋亡。

Objective To explore the mechanism of the targeted inhibition of the formin like 2(FMNL2)medicated Notch signaling pathway by MicroRNA-145-5p(miR-145-5p)in regulating the proliferation and apoptosis of oral squamous cell carcinoma(OSCC)cells.Methods The targeting regulatory relation between miR-145-5p and FMNL2 was confirmed firstly.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were utilized to detect the expression of miR-145-5p,FMNL2 and the Notch signaling pathway related factors Notch 1 and Hes1 in the cancer tissues and their adjacent tissues.The expression of miR-145-5p and FMNL2 in OSCC cells were intervened with and then the cells were divided into different groups.qRT-PCR and Western blot were adopted to detect the expression of miR-145-5p,FMNL2,the Notch signaling pathway related factors and the apoptosis-related factors Bax and Bcl-2 in each group.The methyl thiazolyl tetrazolium(MTT)method was used to detect the cell proliferation activity,the Transwell assay was to detect the invasion ability,and the flow cytometry assay was to measure the cell apoptosis.Results FMNL2 was confirmed as the target gene of miR-145-5p.Compared with the adjacent tissues,the miR-145-5p expression was decreased,the FMNL2 expression was increased and the Notch signaling pathway was activated in the cancer tissues in the tissue experiment(P<0.05).In the cell experiment,the miR-145-5p overexpression or the FMNL2 silencing inhibited the expression of the Notch signaling pathway related factors,increased the Bax expression and decreased the Bcl-2 expression while inhibiting the proliferation and invasion of OSCC cells and inducing the cell apoptosis(P<0.05).The contrary changes were observed in the miR-145-5p inhibition group.Conclusion The targeted inhibition of FMNL2 by miR-145-5p can inhibit the proliferation and invasion of OSCC cells and promote the cell apoptosis by inhibiting the Notch signaling pathway activation.