白癜风患者Folliculin蛋白调控黑素细胞趋化因子表达的研究

Regulation of melanocyte chemokine expression by folliculin protein in vitiligo patients

目的探讨人肿瘤抑制因子Folliculin(FLCN)对白癜风发病相关免疫因子介导的黑素细胞(MC)趋化因子表达的调控。方法收集2019年1—4月杭州市第三人民医院皮肤科接受自体黑素细胞移植白癜风患者MC,Western印迹法分析正常MC、白癜风患者MC、免疫因子诱导MC中FLCN蛋白表达差异;通过短发卡RNA(shRNA)构建FLCN shRNA慢病毒并转染正常黑素细胞(FLCN shRNA MC)干扰沉默FLCN基因的表达,ELISA方法检测免疫因子对FLCN shRNA MC中趋化因子的影响。结果Westem印迹结果显示,白癜风患者MC中FLCN高表达;免疫因子可刺激正常MC中FLCN表达显著升高(t=1.27;P<0.001),也可诱导趋化因子CXCL10和CCL20显著升高(t分别为104.53、60.21;均P<0.001);FLCN shRNA MC的FLCN表达显著降低(F=1.95;P<0.001),且显著抑制免疫因子诱导的CXCL10和CCL20高表达(F=93.68、74.10;均P<0.001)。结论免疫因子刺激MC中CXCL10、CCL20高表达,与白癜风发病密切相关,FLCN是参与免疫因子诱导MC趋化因子表达的关键蛋白。

Objective To investigate the effect of human tumor suppressor folliculin(FLCN)on the expression of melanocyte chemokines(MC)mediated by immune factors in vitiligo.Methods The MC of vitiligo patients that received autologous melanocyte transplantation in the Department of Dermatology,Hangzhou Third People′s Hospital from January to April 2019 were collected.The blister fluid of the white spot and the normal part was taken.Western blot was used to analyze the expression difference of MC and FLCN protein in normal,vitiligo patients and that induced by immune factors;FLCN shRNA lentivirus was constructed by shRNA and transfected into normal MC(FLCN shRNA MC)to interfere with the expression of silenced FLCN gene.The effect of immune factors on chemokines in FLCN shRNA MC was detected by ELISA.Results The results of Western blot showed that FLCN protein was highly expressed in melanocytes of vitiligo patients,immune factors stimulated FLCN protein expression in normal melanocytes significantly increased(t=1.27;P<0.001),chemokine CXCL10 and CCL20 also significantly increased(t=104.53 and 60.21,respectively;P<0.001).The expression of FLCN in FLCN shRNA MC was significantly decreased(F=1.95,P<0.001);and the high expression of CXCL10 and CCL20 induced by immune factors was significantly inhibited(F=93.676 and 74.096,all P<0.001).Conclusions Immune factors can stimulate the expression of CXCL10 and CCL20,which are closely related to vitiligo,while FLCN is a key protein involved in immune factors inducing melanocyte chemokine expression.