摘要
目的:探讨miR-196-5p对皮肤鳞状细胞癌(CSCC)增殖、侵袭和EMT的影响及其潜在的作用机制。方法:利用实时荧光定量PCR(RT-qPCR)检测miR-196-5p在癌旁正常组织、CSCC组织、A431细胞和HaCaT细胞中的表达水平。过表达或下调miR-196-5p在A431细胞中的表达后,通过CCK-8实验检测miR-196-5p对A431细胞增殖能力的影响;细胞侵袭实验检测miR-196-5p对A431细胞侵袭能力的影响;Western blot检测miR-196-5p对A431细胞内E-cadherin、N-cadherin表达的影响。生物信息学软件TargetScan预测miR-196-5p和HOXA5的作用位点,双荧光素酶报告基因实验分析miR-196-5p和HOXA5之间的关系。上调HOXA5在A431细胞内表达后检测其对A431细胞增殖、侵袭和EMT的影响。Western blot分析下调HOXA5对JAK/STAT通路的影响。AG490作用细胞后,分析下调HOXA5对A431细胞增殖、侵袭和EMT的影响。结果:与癌旁正常组织相比,CSCC组织中miR-196-5p表达水平明显升高(t=6.32,P=0.024),HOXA5表达明显降低(t=7.60,P=0.017)。与HaCaT细胞相比,A431细胞中miR-196-5p表达明显升高(t=18.58,P=0.003),HOXA5表达明显降低(t=19.72,P=0.003)。过表达miR-196-5p促进A431细胞增殖、侵袭与EMT,下调miR-196-5p则起到相反作用;miR-196-5p靶向且负调控HOXA5;过表达HOXA5抑制A431细胞增殖、侵袭与EMT;下调HOXA5激活细胞内JAK/STAT通路。AG490作用细胞后下调HOXA5对A431细胞增殖、侵袭与EMT的促进作用。结论:miR-196-5p靶向HOXA5激活JAK/STAT通路促进CSCC的增殖、侵袭和EMT。
Objective:To investigate the effects of miR-196-5 p on the proliferation, invasion and EMT of skin squamous cell carcinoma and its potential mechanism. Methods: Quantitative real-time PCR(RT-qPCR) was used to detect the expression of miR-196-5 p in normal paracancer tissues, skin squamous cell carcinoma tissues, A431 cell and HaCaT cell. After the expression of miR-196-5 p was overexpressed or down-regulated, the effects of miR-196-5 p on the proliferation ability of A431 cells were detected by CCK-8 experiment. The effects of miR-196-5 p on the invasion ability of A431 cells were detected by transwell. Western blot was used to detect the effects of miR-196-5 p on the expression of E-cadherin and N-cadherin in A431 cells. TargetScan predicted the action sites of miR-196-5 p and HOXA5. The relationship between miR-196-5 p and HOXA5 was analyzed by dual luciferase reporter gene assay. After up-regulating HOXA5 expression in A431 cells, the effects of HOXA5 on A431 cell proliferation, invasion and EMT were detected. The effects of down-regulation of HOXA5 on the JAK/STAT pathway was detected by western blot. After AG490 was applied to cells, the effects of down-regulation of HOXA5 on A431 cell proliferation、invasion and EMT were detected.Results:The expression level of miR-196-5 p in cutaneous squamous cell carcinoma tissues was significantly higher than that in paracancer tissues(t=6.32,P=0.024), HOXA5 expression significantly decreased(t=7.60,P=0.017). Compared with HaCaT cells, miR-196-5 p expression was significantly increased(t=18.58,P=0.003), HOXA5 expression significantly decreased in A431 cells(t=19.72,P=0.003). Overexpression of miR-196-5 p promoted proliferation, invasion and EMT of A431 cells, while down-regulation of miR-196-5 p had an opposite effect. miR-196-5 p targeted and negatively regulated HOXA5.Overexpression of HOXA5 inhibited the proliferation, invasion and EMT of A431 cells. Down-regulation of HOXA5 activated the JAK/STAT pathway in A431 cell. AG490 downregulated the promotion of HOXA5 on A
作者
张永红
代雪莲
李存涛
张玉红
ZHANG Yonghong;DAI Xuelian;LI Cuntao;ZHANG Yuhong(Department of Dermatology,Zhengzhou Central Hospital,Zhengzhou University,Zhengzhou 450007,China)
出处
《皮肤性病诊疗学杂志》
2021年第4期254-260,267,共8页
Journal of Diagnosis and Therapy on Dermato-venereology
基金
河南省科技发展计划项目(202102310397)。
