siRNA沉默STAT3表达对IL-17诱导人角质形成细胞HaCaT增殖的影响  

Effect and mechanism of STAT3 gene silence by siRNA on proliferation of human keratinocyte HaCaT cells

作　　者：徐媛媛[1] 于雅洁[1] 刘佳[1] 耿龙[1] 高兴华[1] 

机构地区：[1]中国医科大学附属第一医院皮肤性病科,沈阳110001

出　　处：《解剖科学进展》2017年第4期345-348,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81402595)

摘　　要：目的通过体外靶向沉默STAT3基因转录,探讨STAT3抑制对IL-17诱导人角质形成细胞HaCaT增殖能力的影响及其机制。方法采用IL-17A(80ng/ml)刺激体外培养的HaCaT细胞,采用小干扰RNA(Small interfering RNA,siRNA)干扰技术体外沉默人角质形成细胞HaCaT中STAT3基因转录,采用qRT-PCR和Western blot分别检测细胞内STAT3 mRNA,STAT3和pSTAT3蛋白的表达变化,证实基因沉默效果,CCK-8法检测HaCaT细胞增殖能力的变化,Western blot检测STAT3的靶分子survivin及cyclin D1表达的变化,初步探讨STAT3抑制对IL-17诱导Hacat增殖调控的分子机制。结果转染STAT3 siRNA质粒可显著抑制HaCaT细胞中STAT3 mRNA表达,STAT3和pSTAT3蛋白表达(P<0.05)。STAT3抑制后,IL-17诱导HaCaT增殖能力显著降低(P<0.05),STAT3的靶分子survivin及cyclin D1表达水平显著降低(P<0.05)。结论 siRNA干扰靶向抑制STAT3表达可抑制IL-17诱导HaCaT增殖,可能与其抑制survivin及cyclin D1表达有关。Objective To study the effect and mechanism of STAT3 gene silence by siRNA by siRNA on the proliferation ability of human keratinoeyte cell line HaCaT induced by IL-17. Methods HaCaT cells cultured in vitro was pre-stimulated by IL-17A （80ng/ml） for 24h. STAT3 in HaCaT cells was silenced by siRNA. 48h later, qRT-PCR and Western blot were used to detect the expression of STAT3 mRNA and proteins, and pSTAT3 proteins in HaCaT cells, CCK8 assay was performed to evaluate the proliferation of HaCaT cells, and Western blot was used to detect the expression of target gene survivin and cyclin D1. Results The expression levels of STAT3 mRNA, STAT3 and pSTAT3 proteins in HaCaT cells pre-stimulated by IL-17A were all decreased. The cell proliferation and the expression level of survivin and cyclin D1 in HaCaT cells pre-stimulated by IL-17A were all obviously decreased in the transfected group than in control group （P 〈 0.05 ） . Conclusion STAT3 siRNA inhibited the proliferation of HaCaT cells pre-stimulated by IL-17A, which may be related to its inhibitive effect on the expression of survivin and cyclin D1.

关 键 词：银屑病 STAT3 siRNA IL-17 增殖 HaCaT细胞 

分 类 号：R758.4[医药卫生—皮肤病学与性病学]