微小RNA-3651过表达通过介导核因子κB信号通路抑制人舌癌细胞CAL27的生长与侵袭

microRNA-3651 overexpression inhibiting the growth and invasion of human tongue cancer CAL27 cells by mediating nuclear factorκB signaling pathway

目的探讨微小RNA(miR)-3651过表达通过介导核因子(NF)-κB信号通路抑制人舌癌细胞CAL27生长与侵袭的机制。方法将对数生长期CAL27细胞分为miR-3651 mimic组、mimic-NC组和对照组。采用qRT-PCR检测转染后细胞miR-3651水平,CCK-8检测细胞活力,Transwell检测细胞侵袭能力,划痕实验检测细胞迁移能力,光学显微镜下观察细胞上皮间质转化(epithelial-mesenchymal transition,EMT)的形成,Western blot检测E钙粘蛋白、N钙粘蛋白、波形蛋白、NF-κB p65和p-NF-κB p65蛋白表达。结果mimic-NC组与对照组细胞miR-3651水平、细胞活力、侵袭细胞数、细胞迁移率和细胞形态均无明显差异(P>0.05),E钙粘蛋白、N钙粘蛋白、波形蛋白和p-NF-κB p65/NF-κB p65蛋白表达均无明显差异(P>0.05);与mimic-NC组相比,miR-3651 mimic组细胞miR-3651水平升高(P<0.05),培养的细胞第2、3、4天的活力降低(P<0.05),侵袭细胞数和细胞迁移率降低(P<0.05),细胞E钙粘蛋白表达升高(P<0.05),N钙粘蛋白、波形蛋白和p-NF-κB p65/NF-κB p65蛋白表达下降(P<0.05),显微镜下观察到EMT形成减少。结论过表达miR-3651可以抑制CAL27细胞的生长和侵袭,其作用机制可能与抑制NF-κB信号通路磷酸化有关。

Objective To explore the mechanism of microRNA(miR)-3651 overexpression on inhibiting the growth and invasion of human tongue cancer CAL27 cells by mediating nuclear factor(NF)-κB signaling pathway.Methods CAL27 cells in logarithmic growth phase were divided into a miR-3651 mimic group,a mimic-NC group and a control group.The level of miR-3651 in each group after transfection was detected by fluorescent quantitative real-time polymerase chain reaction(qRT-PCR).The cell vitality in each group was detected by cell counting kit-8(CCK-8).The invasion ability of cells in each group was detected by Transwell.The migration ability of cells in each group was detected by scratch assay.The formation of epithelial-mesenchymal transition(EMT)in each group was observed under light microscope.The expression of E-cadherin,N-cadherin,Vimentin,NF-κB p65 and p-NF-κB p65 was detected by Western Blot.Results There was no statistically significant difference in miR-3651 level,activity of cells vitality,number of invasion cells,cells migration rate and cells morphology between mimic-NC group and control group(P>0.05),and there was no significant difference in the expression of E-cadherin,N-cadherin,Vimentin and p-NF-κB p65/NF-κB p65 proteins(P>0.05).Compared with the mimic-NC group,in the miR-3651 mimic group,miR-3651 level was significantly increased(P<0.05),cells vitality were significantly decreased at 2d,3d,4d after cell culture(P<0.05),number of invasion cells and cells migration rate were significantly decreased(P<0.05),expression of E-cadherin protein was significantly increased(P<0.05),expression of Ncadherin,Vimentin and p-NF-κB p65/NF-κB p65 proteins were significantly decreased(P<0.05),and EMT formation under microscope was decreased.Conclusions miR-3651 overexpression can inhibit the growth and invasion of CAL27 cells,and its action mechanism may be related to the inhibiting the phosphorylation of NF-κB signaling pathway.