慢病毒介导的RNA干扰FOXC2表达对皮肤鳞状细胞癌A431细胞功能影响

目的:探讨慢病毒介导的RNA干扰叉头框C2(FOXC2)表达对皮肤鳞状细胞癌A431细胞增殖、侵袭和凋亡的影响及其机制.方法:采用实时荧光定量聚合酶链反应(PCR)和免疫印迹法检测人正常永生化上皮Hecate细胞和皮肤鳞状细胞癌A431细胞中FOXC2 mRNA和蛋白的表达水平.将体外培养的A431细胞分为对照组(正常...

Effect of lentivirus mediated RNA interference on FOXC2 expression on the function of cutaneous squamous cell carcinoma A431 cells

Objective:To investigate the effect of lentivirus mediated RNA interference expression of FOXC2 on proliferation,invasion and apoptosis of cutaneous squamous cell carcinoma A431 cells and mechanism.Methods:The expression of FOXC2 mRNA and protein in human normal immortalized epithelial Hecate cells and cutaneous squamous cell carcinoma A431 cells were detected by real-time fluorescence quantitative PCR and Western blot.A431 cells cultured in vitro were divided into control group(normal culture),NC shRNA group(LV-NC-shRNA lentivirus infection)and FOXC2-shRNA group(LV-FOXC2-shRNA lentivirus infection).Cell viability was detected by MTT method.Cell clonogenesis was detected by plate clonogenesis test.Cell invasion was detected by Transwell chamber test.Cell apoptosis was detected by flow cytometry and the expression levels of AKT,p-AKT,CyclinD1,MMP-9 and Bcl-2 were measured by Western blotting.Results:Compared with Hecate cells,the expression of FOXC2 mRNA and protein in A431 cells increased significantly(P<0.05).Compared with the control group,there was no significant difference in FOXC2 mRNA expression level,cell survival rate,invasion number,clone formation rate,apoptosis rate and the expression levels of FOXC2,p-AKT,CyclinD1,MMP-9,Bcl-2 proteins in NC-shRNA group(P>0.05).Compared with the control group or NC-shRNA group,FOXC2 mRNA expression level,cell survival rate,invasion number and the expression levels of FOXC2,p-AKT,CyclinD1,MMP-9 and Bcl-2 in FOXC2-shRNA group were significantly decreased,while the apoptosis rate was significantly increased(P<0.05).Conclusion:The inhibition of FOXC2 expression can inhibit the proliferation,invasion and promote apoptosis of cutaneous squamous cell carcinoma A431 cells,and its mechanism may be related to the inhibition of AKT signaling pathway activation.