RNA干扰CLEC2B基因沉默对淋巴细胞可溶性白介素2受体mRNA表达的影响  

Effects of RNA Interfering CLEC2B Gene Silence on the Expression of sIL-2R mRNA

作　　者：张峻岭[1,2] 柳君如[3] 徐士福[4] 程琳 马秀亮[1,2] 

机构地区：[1]天津市中医药研究院附属医院 [2]天津市长征医院皮肤色素科,300120 [3]中国人民解放军第404医院皮肤科 [4]浙江大学医学院附属儿童医院 [5]河北省石家庄市妇幼保健院

出　　处：《中国全科医学》2013年第27期3199-3201,共3页Chinese General Practice

基　　金：天津市自然科学基金(09JCYBJC09600)

摘　　要：目的探讨RNA干扰淋巴细胞中CLEC2B基因沉默后,对淋巴细胞增殖及其细胞因子可溶性白介素2受体(sIL-2R)mRNA表达的调节作用,揭示CLEC2B基因参与白癜风发病的免疫学机制。方法人Jurkat淋巴瘤细胞株细胞培养后,分别应用RNA干扰技术转染3条CLEC2B siRNA(沉默1组、沉默2组、沉默3组),转染scrambled siRNA(非沉默对照组)及不做任何处理(空白对照组)。应用实时荧光定量反转录聚合酶链式反应(RT-PCR)方法分析CLEC2B基因沉默效率,MTT法检测CLEC2B基因沉默后淋巴细胞增殖率(A值),荧光定量PCR方法检测sIL-2R mRNA表达情况。结果转染CLEC2B siRNA后,各CLEC2B沉默组较非沉默对照组淋巴细胞CLEC2B mRNA表达下降,差异均有统计学意义(P<0.05)。各CLEC2B沉默组siRNA序列对CLEC2B基因沉默效率不等,差异有统计学意义(P<0.01);其中沉默2组siRNA沉默效率最高,与沉默1组和沉默3组比较,差异均有统计学意义(P<0.05)。CLEC2B基因沉默后,空白对照组、非沉默对照组、CLEC2B沉默组淋巴细胞增殖率比较,差异无统计学意义(P>0.05)。3组sIL-2R mRNA相对表达水平比较,差异有统计学意义(P<0.05);其中CLEC2B沉默组较非沉默对照组降低,差异有统计学意义(P<0.05)。结论 CLEC2B基因表达沉默后可下调淋巴细胞sIL-2R mRNA的转录表达,提示CLEC2B可能通过调节淋巴细胞产生的细胞因子水平参与白癜风的免疫发病机制。Objective To observe the effect of RNA interfering CLEC2B gene silence on the proliferation of lymphocytes and expression of sIL-2R mRNA in order to reveal the immunity mechanism of CLEC2B in vitiligo.Methods RNA interfering was used in the transfection of three CLEC2B siRNA(silence group one,silence group two and silence group three) and scrambled siRNA(non-silence control group) after cell culture of lymphoma cell lines.The cell line without any treatment was involved as blank control group.Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR) method was used to analyze the CLEC2B efficiency of gene silencing,MTT method was used to detect CLEC2B lymphocyte proliferation rate after gene silencing and fluorescence quantitative polymerase chain reaction(PCR) method was used to detect the expression of sIL-2R mRNA.Results After transfection of CLEC2B siRNA,the lymphocyte CLEC2B mRNA expression in each silence group was significantly reduced compared with non-silence group(P < 0.05).The CLEC2B gene silencing efficiency by siRNA sequences in each silence group showed statistically significant differences(P < 0.01).Among them,the silence group two showed the highest silencing efficiency compared with the silence group one and silence group three(P < 0.05).After gene silence of CLEC2B,the lymphocyte proliferation rate between the blank control group,non-silence control group and each CLEC2B silence group showed no statistically significant difference(P > 0.05).The expression of cytokines sIL-2R mRNA between the blank control group,non-silence control group and each CLEC2B silence group showed statistically significant differ-ences( P < 0. 05). Among them,the expression of cytokines sIL- 2R mRNA in CLEC2B silence groups was significantly reduced compared with the non- silence control group( P < 0. 05). Conclusion CLEC2B gene silence can down- regulate the transcription and expression of cytokine sIL- 2R mRNA,indicating that CLEC2B may be involved in the immune pathogenesis of vitilig

关 键 词：白癜风 淋巴细胞 CLEC2B 受体 白介素2 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]