LncRNA DSCAM-AS1在寻常型银屑病皮损组织中的表达及其对IL-22诱导的角质形成细胞增殖和凋亡的影响  

Expression of LncRNA DSCAM-AS1 in skin lesions of psoriasis vulgaris and its effects on IL-22-induced keratinocyte proliferation and apoptosis

作　　者：李旭阳[1] 郑云鹏[1] 金芳草[1] 尹光文[1] LI Xuyang;ZHENG Yunpeng;JIN Fangcao;YIN Guangwen(Department of Dermatology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院皮肤科,郑州450052

出　　处：《郑州大学学报（医学版）》2022年第5期663-669,共7页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省重点研发与推广专项(科技攻关)项目(222102310458)。

摘　　要：目的:探讨LncRNA唐氏综合征细胞黏附分子反义RNA 1(DSCAM-AS1)在寻常型银屑病皮损组织中的表达,并观察其对白细胞介素22(IL-22)诱导的角质形成细胞增殖和凋亡的影响及可能机制。方法:采用qRT-PCR法分别检测37例寻常型银屑病患者皮损组织、37例行整形手术者正常皮肤组织以及IL-22(100μg/L)刺激24 h的角质形成细胞HaCaT、未处理HaCaT细胞中DSCAM-AS1和miR-199b-3p的表达水平。体外培养HaCaT,分别转染DSCAM-AS1小干扰RNA、miR-199b-3p模拟物或共转染DSCAM-AS1小干扰RNA与miR-199b-3p抑制剂后,用100μg/L的IL-22干预24 h,分别设相应阴性对照,并以未处理细胞为空白对照。采用qRT-PCR法检测细胞中DSCAM-AS1和miR-199b-3p表达,CCK-8法检测细胞增殖,Annexin V-FITC/PI双染法检测细胞凋亡,Western blot法检测CyclinD1、Bax、Bcl-2及IL-6R/STAT3信号通路相关蛋白IL-6R和STAT3蛋白的表达。利用双荧光素酶报告实验验证DSCAM-AS1和miR-199b-3p的靶向调控关系。结果:与正常皮肤组织比较,寻常型银屑病皮损组织中DSCAM-AS1表达增加,miR-199b-3p表达降低(P<0.05);HaCaT细胞经IL-22刺激后,DSCAM-AS1表达增加,miR-199b-3p表达降低(P<0.05)。敲减DSCAM-AS1或过表达miR-199b-3p可降低IL-22刺激的HaCaT细胞增殖能力及CyclinD1、Bcl-2、IL-6R和STAT3蛋白表达,提高细胞凋亡率及Bax蛋白表达(P<0.05)。DSCAM-AS1靶向结合并负调控miR-199b-3p。敲减miR-199b-3p可部分逆转DSCAM-AS1沉默对IL-22刺激的HaCaT细胞增殖、凋亡及IL-6R/STAT3信号通路的影响(P<0.05)。结论:DSCAM-AS1在寻常型银屑病皮损组织及IL-22刺激的HaCaT细胞中表达增加,敲减其表达可抑制IL-22刺激的HaCaT细胞增殖,并促进细胞凋亡,其机制与靶向miR-199b-3p并阻断IL-6R/STAT3信号通路有关。Aim:To investigate the expression of LncRNA Down syndrome cell adhesion molecule-AS1(DSCAM-AS1)in skin lesions of psoriasis vulgaris,and observe its effects on the proliferation and apoptosis of keratinocytes induced by interleukin 22(IL-22)and its possible mechanism.Methods:The skin lesion tissue of 37 patients with psoriasis vulgaris and the normal skin tissue of 37 patients undergoing plastic surgery were collected.Keratinocytes HaCaT were treated with IL-22(100μg/L)for 24 hours,and those without treatment were used as control.qRT-PCR was used to detect the expression levels of DSCAM-AS1 and miR-199b-3p in the above samples and cells.HaCaT cells were cultured in vitro and transfected with DSCAM-AS1 small interfering RNA,miR-199b-3p mimics or co-transfected with DSCAM-AS1 small interfering RNA and miR-199b-3p inhibitor,respectively,and then 100μg/L IL-22 was used to intervene the transfected HaCaT cells for 24 hours,with corresponding negative controls and cells not treated as control.qRT-PCR was used to detect the expressions of DSCAM-AS1 and miR-199b-3p in cells;CCK-8 was used to detect cell proliferation;Annexin V-FITC/PI double staining was used to detect cell apoptosis;Western blot was used to detect the protein expressions of CyclinD1,Bax,Bcl-2,IL-6R and STAT3.The dual luciferase reporter gene experiment verified the targeted regulation relationship between DSCAM-AS1 and miR-199b-3p.Results:Compared with normal skin tissue,the expression of DSCAM-AS1 in psoriasis lesions increased,while the expression of miR-199b-3p decreased(P<0.05).After being stimulated by IL-22,the expression of DSCAM-AS1 in HaCaT cells increased,while the expression of miR-199b-3p decreased(P<0.05).Knockdown of DSCAM-AS1 or overexpression of miR-199b-3p inhibited the proliferation of IL-22 stimulated HaCaT cells and the protein expressions of CyclinD1,Bcl-2,IL-6R and STAT3,while increased the rate of apoptosis and the protein expression of Bax(P<0.05).DSCAM-AS1 negatively regulated the expression of miR-199b-3p.Knockdowning miR-199

关 键 词：银屑病 唐氏综合征细胞黏附分子反义RNA 1 miR-199b-3p 角质形成细胞 细胞增殖 细胞凋亡 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]