干扰硒蛋白K对小鼠T淋巴细胞钙流和白介素2受体表达的影响  

Effect of selenoprotein K silence on intracellular free Ca^(2+) and IL-2R expression in mouse T lymphocytes in vitro

作　　者：杨永平 李瑞敏[2] 李晓霞[3] 金玉娟[3] 王超[2] 张仁利[2] 

机构地区：[1]深圳市龙岗区坪地预防保健所,广东深圳518117 [2]深圳市疾病预防控制中心,广东深圳518052 [3]深圳市龙岗区疾病预防控制中心,广东深圳518116

出　　处：《中国热带医学》2016年第10期961-964,共4页China Tropical Medicine

基　　金：深圳市科技创新委项目(No.JCYJ20150402141723407)

摘　　要：目的探讨硒蛋白K(Selenoprotein K,Sel K)基因沉默(RNA silence,RNAi)对小鼠T淋巴细胞内钙流和白介素2受体(Interleukin-2 Receptor,IL-2R)分泌的影响。方法制作小鼠T淋巴细胞干扰载体p GPU-Sel K,用脂质体Lipofectin 2000将干扰质粒和对照质粒转染小鼠T淋巴细胞,Real time PCR和Western Blot分别检测核酸和蛋白水平Sel K的表达,流式细胞仪检测细胞内钙离子浓度,Real time PCR检测IL-2R的分泌。结果与转染的对照组质粒p GPU-NC比较,转染p GPU-Sel K载体的细胞Sel K在m RNA和蛋白水平表达分别显著降低了42.37%(P<0.01)和45.44%(P<0.01)。另外干扰组整体细胞内钙离子浓度显著低于对照组(P<0.05),细胞分泌IL-2R水平也显著降低(P<0.05)。结论干扰载体p GPU–Sel K可以下调小鼠T淋巴细胞Sel K的表达,抑制细胞的钙流和IL-2R的分泌,从而可能改变小鼠T淋巴细胞增殖分化。Objective To determine the effect of silencing selenoprotein K（Sel K） gene on intracellular free Ca^2＋ and theexpression of IL-2R in mouse T lymphocytes. Methods The RNA interference vector of mouse Selk gene was constructed,and then the interference and control vectors were resepectively transfected into mouse T cells using Lipofectin 2000. Them RNA and protein levels of Sel K were detected by real-time PCR and Western blotting, respectively. Intracellular free Ca^2＋ was measured by flow cytometry, and the expression of IL- 2R in the supernatant was detected by real-time PCR. Results The expression of Sel K at m RNA and protein levels was significantly decreased by 42.37%（P〈0.01） and 45.44%（P〈0.01） inthe RNAi vector group, respectively, when compared with the control vector group. Sel K silence also obviously decreased theintracellular free Ca^2＋ and the expression of IL- 2R（P〈0.05）. Conclusion RNA interference vector p GPU- Sel K down-regulates the expression of Sel K in mouse T cells, inhibits intracellular free Ca2 ＋and the secretion of IL- 2R, and thusmodulates the proliferation and differentiation of the cells.

关 键 词：硒蛋白K T淋巴细胞 钙流 白介素2受体 

分 类 号：R446.63[医药卫生—诊断学]