IRF1受到miR-23a调控抑制舌鳞癌细胞增殖并促进凋亡

IRF1 Regulated by MiR-23a to Inhibit Proliferation and Promote Apoptosis in Tongue Squamous Carcinoma Cells

目的:研究IRF1基因对舌鳞癌细胞增殖和凋亡的影响及其上游受到miR-23a调控的作用机制。方法:通过Real-time PCR在舌鳞癌组织中检测IRF1基因的表达水平。构建IRF1的过表达质粒,通过MTT实验和TUNEL实验分别检测IRF1对舌鳞癌细胞增殖和凋亡的影响。生物信息学预测以及荧光报告载体实验验证IRF1是miR-23a的直接作用的靶基因。进一步检测舌鳞癌组织中miR-23a的表达水平以及封闭miR-23a表达后,舌鳞癌细胞增殖和凋亡的改变。结果:在舌鳞癌组织中IRF1表达水平较癌旁正常组织明显降低。在舌鳞癌细胞中过表达IRF1,细胞的生长活性明显降低,而紫杉醇诱导的凋亡指数升高。TargetScan预测miR-23a可以与IRF1的3‘UTR相结合。经证实miR-23a可以直接与IRF1的3‘UTR结合,进而调控IRF1的表达。miR-23a在舌鳞癌组织中表达异常升高,并且封闭miR-23a的表达后,舌鳞癌细胞的生长活性明显降低,而紫杉醇诱导的凋亡指数升高,与过表达IRF1所得结果一致。结论:IRF1受到miR-23a的调控抑制舌鳞癌细胞的生长活性并促进其凋亡。IRF1在舌鳞癌发生发展中是抑癌基因,而miR-23a作为IRF1的上游调控因子,是舌鳞癌发生发展中的致癌基因。

Objective:To study the effect of IRF1 gene on proliferation and apoptosis of tongue squamous carcinoma cells and its upstream regulation mechanism by miR-23a.Methods:The expression level of IRF1 gene was detected by Real-time PCR in tongue squamous carcinoma tissues.The overexpression plasmid of IRF1 was constructed,and the effects of IRF1 on proliferation and apoptosis of tongue squamous carcinoma cells were examined by MTT assay and TUNEL assay,respectively.Bioinformatic predictions as well as fluorescent reporter vector assays verified that IRF1 was a direct target gene for miR-23a.The expression level of miR-23a in tongue squamous carcinoma tissues and the alteration of proliferation and apoptosis of tongue squamous carcinoma cells after the expression of miR-23a was further examined.Results:IRF1 expression levels were significantly lower in tongue squamous carcinoma tissues compared to paraneoplastic normal tissues.Overexpression of IRF1 in tongue squamous carcinoma cells resulted in a significant decrease in cell growth activity and an increase in paclitaxel-induced apoptosis index.TargetScan predicted that miR-23a could bind to the 3'UTR of IRF1.It was confirmed that miR-23a could directly bind to the 3'UTR of IRF1 and thus regulate the expression of IRF1.The expression of miR-23a was abnormally elevated in tongue squamous carcinoma tissues,and after the expression of miR-23a was blocked,the growth activity of tongue squamous carcinoma cells was significantly decreased,while the apoptosis index induced by paclitaxel was increased,which was consistent with the results obtained from overexpression of IRF1.Conclusion:IRF1 is regulated by miR-23a to inhibit the growth activity and promote apoptosis of tongue squamous carcinoma cells.IRF1 is a tumor suppressor gene in the occurrence and development of tongue squamous cell carcinoma,while miR-23a,as an upstream regulator of IRF1,is also an oncogene of tongue squamous cell carcinoma.