miR-21-3p靶向STAT3促进银屑病角质形成细胞增殖的研究  

miR-21-3p targeting STAT3 promotes proliferation of psoriatic keratinocytes

作　　者：王慧琴[1] 谢敏欣 丁媛[1] 吴卫东[1] WANG Huiqin;XIE Minxin;DING Yuan;WU Weidong(Department of Dermatology and Venereology,The People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China;Department of Dermatology,Korla Hospital of the Second Division of Xinjiang Production and Construction Corps,Kuerle 841000,Xinjiang,China)

机构地区：[1]新疆维吾尔自治区人民医院皮肤性病科,新疆乌鲁木齐830001 [2]新疆生产建设兵团第二师库尔勒医院皮肤科,新疆库尔勒841000

出　　处：《西部医学》2024年第5期661-666,共6页Medical Journal of West China

基　　金：新疆维吾尔自治区自然科学基金项目(2019D01C130);新疆维吾尔自治区自然科学基金重点项目(2022D01D52)。

摘　　要：目的探讨miR-21-3p及其靶蛋白信号转导子和转录激活子3蛋白(STAT3)在寻常型银屑病发生过程中角质形成细胞的作用意义。方法对皮肤组织中STAT3蛋白表达情况的检测选择免疫组织化学法,对健康人群皮肤组织及银屑病患者皮损组织miR-21-3p表达量的检测选择实时荧光定量PCR法,靶基因STAT3和miR-21-3p关系的检测选择双荧光素酶报告基因。化学合成miR-21-3p模拟物、抑制剂,瞬时转染HaCaT细胞,对不同分组细胞进行CCK8、流式细胞凋亡实验,探究分析miR-21-3p调控细胞表型主要作用。对于转染细胞中STAT3蛋白表达情况,选择Western Blot检测。结果寻常型银屑病患者皮肤组织中miR-21-3p呈高表达(P<0.05)。STAT3免疫组化阳性表达定位于细胞浆,在银屑病组患者皮损组织阳性表达率为82.22%(37/45)高于正常皮肤组织(P<0.01)。双荧光素酶报告基因显示miR-21-3p与STAT3存在结合,呈靶向关系。miR-21-3p mimics促进HaCaT细胞增殖活性,抑制细胞凋亡;miR-21-3p inhibitor抑制HaCaT细胞的增殖能力,促使细胞趋向凋亡。miR-21-3p转染HaCaT过表达可对STAT3蛋白表达量产生促进,相反则降低(F=48.632,P<0.01)。结论miR-21-3p呈现正调控关系,能够靶向调控STAT3;两者共同作用参与并加重银屑病的病程,并抑制患者细胞凋亡,调节角质形成细胞增殖活性。Objective To investigate the role of miR-21-3p and its target protein signal transducer and activator of transcription 3 protein(STAT3)in keratinocytes during the pathogenesis of psoriasis vulgaris.Methods Immunohistochemical method was used to detect the expression of STAT3 protein in skin tissue,real-time fluorescent quantitative PCR method was used to detect the expression of miR-21-3p in skin lesions,and double luciferase reporter gene was used to detect the relationship between STAT3 and miR-21-3p.Chemical synthesis of miR-21-3p mimics and inhibitors,transient transfection of HaCaT cells,CCK8 and flow cytometry apoptosis experiments were carried out on different groups of cells to explore and analyze the main role of miR-21-3p in regulating cell phenotype.The expression of STAT3 protein in transfected cells was detected by Western blot.Results With real-time fluorescent quantitative PCR,the expression level of miR-21-3p was higher in lesions of psoriasis vulgaris than in the beside-lesional tissue and normal skin tissue,and differences was statistical significance(t=7.009,P<0.01).Immunohistochemical results showed that the positive expression of STAT3 protein was located in the cytoplasm.The positive expression of STAT3 in skin lesions of patients with psoriasis population was 82.22%(37/45),it was higher than the rate of positive expression in the normal group.The two groups of data by chi-square test were statistically significant(F=25.742,P<0.01).Transfected HaCaT cells with miR-21-3p mimic and miR-21-3p inhibitor.The results of CCK8 and flow cytometry showed that miR-21-3p mimics promoted the proliferation of HaCaT cells and inhibited apoptosis.MiR-21-3p inhibitor inhibits the proliferation of HaCaT cells and promotes cell apoptosis.The results of Western blot showed that after miR-21-3p was transfected into HaCaT cells,its overexpression could promote the expression of STAT3 protein,on the contrary,it decreased,and the difference was obvious(F=48.632,P<0.01).The detection results of double luciferase rep

关 键 词：角质形成细胞 miR-21-3p STAT3 表达量 银屑病 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]