miR-203通过靶向调控SOCS3影响人皮肤鳞癌细胞增殖、细胞周期行为及其机制研究

The effect and mechanism of miR-203 on the proliferation and cell cycle behavior of human skin squamous cell carcinoma by SOCS3

目的探究miR-203对人皮肤鳞癌细胞A-431增殖、细胞周期行为的影响。方法实时荧光定量PCR检测27对人皮肤鳞癌/癌旁组织及A-431/Ha Ca T细胞中miR-203的表达。在A-431细胞中过表达miR-203,针对miR-NC组和miR-203 mimic组,通过MTT、流式细胞技术检测miR-203对细胞增殖、细胞周期的影响。生物信息学对miR-203进行靶基因预测,并通过双荧光素酶实验确定miR-203和靶基因是否存在相互作用,Western Blot检测该靶基因在过表达miR-203皮肤鳞癌细胞中表达变化。结果 miR-203在皮肤鳞癌组织及癌细胞A-431中均显著低表达,相对癌旁组织及Ha Ca T细胞分别降低36%及40%。与miR-NC组比较,miR-203 mimic组抑制A-431细胞增殖,导致G1/S进程受阻,S期细胞减少。生物信息学分析表明SOCS3是miR-203直接作用的靶基因,双荧光素酶实验验证了该结果,且过表达miR-203可以明显下调SOCS3的蛋白表达量。结论 miR-203在人皮肤鳞癌中低表达,可以通过负调控靶基因SOCS3抑制鳞癌的生长,具有作为临床治疗靶标分子的潜能。

Objective To study the effect of miR-203 on proliferation and cell cycle behavior of human skin squamous cell carcinoma cell line A-431.Methods The expression of miR-203 in human skin squamous cell carcinoma/adjacent tissues and A-431/HaCaT cells was detected by real-time quantitative PCR.For miR-NC group and miR-203 mimic group,MTT and flow cytometry were used to detect effect of A-431 cell proliferation and cell cycle under overexpression of miR-203.After predicted the target gene of miR-203 by bioinformatics,whether there was an interaction between miR-203and the target gene were validated by dual luciferase experiment.At last,Western Blot was used to test the target gene expression changes of miR-203 over-expression in cutaneous squamous cell carcinoma cells.Results The expression level of miR-203 was reduced in cSCC tissues and A-431cells,which decreased 36%and 40%compared with paracancerous tissue and HaCaT cells.Overexpression of miR-203 can inhibit cell proliferation and result in A-431 cell G1/S progression to be blocked and phase S decreased.Bioinformatics analysis showed that SOCS3 was a target gene of miR-203,and dual luciferase experiment verified the result.Overexpression of miR-203 can down-regulate the expression of SOCS3 protein.Conclusion miR-203 is low-expressed and suppress cSCC by regulating target gene SOCS3,which is the potential target molecule of clinical treatment.