沉默miR-373对喉癌细胞增殖、凋亡能力的影响及作用机制研究

Effect of silencing mir-373 on proliferation and apoptosis of laryngeal cancer cells and its mechanism

目的探究沉默RNA-373(miR-373)对喉癌细胞增殖、凋亡能力的影响及作用机制研究。方法将喉癌细胞分为对照组、过表达组、沉默组3组,并通过细胞转染建立稳定转染的过表达组和沉默组。采用MTT法检测细胞增殖能力,TUNEL法检测细胞凋亡能力,Transwell小室检测细胞侵袭,细胞划痕实验检测细胞迁移,蛋白质印迹法检测各组细胞Wnt/β-catenin信号通路β-连锁蛋白(β-catenin)、c-myc、细胞周期素D1(CyclinD1)、基质金属蛋白酶9(MMP-9)、Bc1-2、Bax蛋白表达量。结果与过表达组相比,沉默组miR-373表达量明显下降(t=15.062,P<0.05)。与过表达组相比,沉默组细胞凋亡率较高,细胞不同时间点增殖率较低(t=31.025、16.453、22.475、29.672,P<0.05);与过表达组相比,沉默组细胞侵袭能力、迁移数均较低(t=35.254、37.205,P<0.05)。与过表达组相比,沉默组β-catenin、c-myc、CyclinD1、MMP-9、Bc1-2蛋白表达量较低,Bax蛋白较高(t=4.218、5.307、4.609、5.005、4.328、3.984,P<0.05)。结论沉默miR-373可能通过促进Bax表达,抑制β-catenin、c-myc、CyclinD1、MMP-9、Bc1-2表达,阻滞Wnt/β-catenin信号通路,从而促进喉癌细胞凋亡,抑制喉癌细胞侵袭、增殖、迁移。

OBJECTIVE To explore the effect of silencing RNA-373(mir-373)on the proliferation and apoptosis of laryngeal cancer cells and its mechanism.METHODS Laryngeal cancer cells were divided into control group,overexpression group and silence group.Stable overexpression group and silence group were established by cell transfection.MTT assay was used to detect cell proliferation,TUNEL method was used to detect the apoptotic ability,cell invasion was detected by Transwell chamber,cell migration was detected by cell scratch test,Western blot was used to detect the expression ofβ-Catenin,c-myc,CyclinD1,MMP-9,bc1-2 and Bax in Wnt/β-catenin signaling pathway.RESULTS Compared with over expression group,the expression of mir-373 in silence group was significantly decreased(t=15.062,P<0.05).Compared with the overexpression group,the apoptosis rate was higher and the proliferation rate was lower in the silencing group at different time points(t=31.025,16.453,22.475,29.672,P<0.05).Compared with overexpression group,the invasion ability and migration number of cells in silencing group were lower(t=35.254,37.205,P<0.05).Compared with overexpression group,the expression levels ofβ-Catenin,c-myc,CyclinD1,MMP-9,bc1-2 protein in silencing group were lower,and Bax protein was higher(t=4.218,5.307,4.609,5.005,4.328,3.984,P<0.05).CONCLUSION Silencing mir-373 may promote apoptosis and inhibit invasion,proliferation and migration of laryngeal cancer cells by promoting Bax expression,inhibiting the expression ofβ-Catenin,c-myc,CyclinD1,MMP-9 and bc1-2,and blocking Wnt/β-catenin signaling pathway.