摘要
目的探讨微小核糖核酸-21(microRNA-21,miR-21)调控其靶蛋白错配修复基因——人mut S同种组蛋白2(h MSH2)在寻常型银屑病发生过程中的表达水平和作用。方法采用实时荧光定量聚合酶链式反应(PCR)法检测miR-21在30例寻常型银屑病患者皮损组织和30例正常皮肤组织中的表达水平。利用免疫组织化学Envision二步法检测30例寻常型银屑病患者与30例正常人群h MSH2蛋白的表达情况。化学合成miR-21模拟物、抑制剂,瞬时转染Ha Ca T细胞以过表达、抑制表达miR-21,采用Western blot法检测转染细胞中h MSH2蛋白的表达。结果miR-21在寻常型银屑病患者皮损组织中的表达高于正常皮肤组织,差异有统计学意义(t=18.596,P<0.01)。h MSH2蛋白在寻常型银屑病患者皮损中的阳性表达率为90.00%,显著高于在正常皮肤组织中的46.67%,差异有统计学意义(χ~2=13.017,P<0.01)。Western blot法检测h MSH2蛋白表达结果显示,过表达miR-21时h MSH2蛋白表达量增加,miR-21抑制表达时h MSH2蛋白表达量降低,差异有统计学意义(F=69.579,P<0.01)。结论miR-21及其靶蛋白h MSH2在寻常型银屑病患者皮肤组织中的表达显著高于正常皮肤组织;miR-21的差异表达可能调控h MSH2蛋白参与寻常型银屑病的发病过程。
Objective To investigate the expression level and significance of microRNA-21 （ miR-21 ） in pathogenic process of psoriasis vulgaris （ PV ） via regulating its target protein-mismatch repair gene- human Mut-S-Homologon-2 （hMSH2） protein. Methods Real time fluorescence quantitative polymerase chain reaction （PCR） was used to detect the expression levels of miR-21 （2-△△Ct） in skin lesion tissues of 30 cases of PV and in normal skin tissues of 30 cases of normal subjects. Immunohistochemical Envision method was used to detect the positive expression rate of hMSH2 protein in skin lesion tissues of 30 cases of PV and in skin tissues of 30 normal subjects. The miR-21 mimic and miR-21 inhibitor were synthesized with chemical method for transient transfection of HaCaT cells in order to induce over-expression or depressive-expression of miR-21. Western blot method was used to detect the expression of hMSH2 protein in transfected cells. Results The expression level of miR-21 in skin lesion tissues of PV patients was significantly higher than that in normal skin tissues （t = 18. 596, P 〈 0. 01 ）. The positive expression rate of hMSH2 protein in skin lesion tissues of PV patients （90. 00% ） was significantly higher than that in normal skin tissue （46. 67% ） （X2 = 13. 017 ,P 〈0. 01 ）. Western blot method showed that compared with negative control group, the expression quantity of hMSH2 protein in transfeced HaCaT cells of miR-21 over-expression increased and decreased in transfeced HaCaT cells of miR-21 depressive-expression （ all P 〈 0.01 ）, and there was significant difference among three groups （ F = 69. 579, P 〈 0.01 ）. Conclusions The expressions of miR-21 and its target protein hMSH2 in skin lesion tissues of PV patients are significantly higher than those in normal skin tissues. The differential expression of miR-21 indicates that miR-21 participates in the pathogenic process of PV maybe through positive regulation of hMSH2 protein.
出处
《中国临床研究》
CAS
2017年第5期582-585,共4页
Chinese Journal of Clinical Research
基金
新疆维吾尔自治区自然科学基金(2015211C213)
作者简介
通讯作者：吴卫东，E-mail：xjwudong@126．com
