miR-194在增龄性胸腺萎缩中的表达及靶基因研究  

Expression and function of miR-194 in thymic epithelial cell with aging

作　　者：易丽君[1] 祝漫琴 周菁[1] 胡清华[1] 黄慧[1] 陈卡[1] 江志军[1] 郭智彬[1] YI Li-Jun;ZHU Man-Qin;ZHOU Jing;HU Qing-Hua;HUANG Hui;CHEN Ka;JIANG Zhi-Jun;GUO Zhi-Bin(Central Laboratory,Jiangxi Provincial Children's Hospital,Nanchang 330006,China)

机构地区：[1]江西省儿童医院中心实验室,南昌330006 [2]江西省儿童医院药剂科,南昌330006

出　　处：《中国免疫学杂志》2018年第3期325-330,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.81460222);江西省卫生计生委科技计划(No.20155564)

摘　　要：目的:检测增龄性胸腺萎缩过程中miR-194与PTPN12的表达水平变化,并分析二者相互作用,阐明其中的分子调节机制。方法:选用C57BL/6小鼠,分为4组:1月龄组、6月龄组、10月龄组和19月龄组,每组6只,雌雄各半。麻醉后取出胸腺组织,用CD45抗体与LS柱吸附洗脱,筛选出胸腺上皮细胞。实时荧光定量PCR与Western blot方法检测随年龄增长,胸腺上皮细胞中miR-194与PTPN12基因的表达变化趋势。体外实验共转染miR-194与PTPN12荧光素酶报告载体到HEK293细胞内,分别于24、48 h后检测自发荧光值。结果:随月龄增长,miR-194表达出现下调趋势(P<0.05),而PTPN12基因表达出现上调趋势(P<0.05),且二者呈负相关性(P<0.05)。体外荧光素酶报告基因结果显示,miR-194与PTPN12基因3'UTR区域发生直接作用,并在48 h结合效率最高。结论:PTPN12是miR-194靶基因之一,参与了增龄性胸腺萎缩过程,是调节胸腺上皮细胞功能的重要因子。Objective:To study the expression and interaction between miR-194 and PTPN12 in the process of age-related atrophy of thymus for clarifying the regulatory mechanism in the process of this disease.Methods:C57BL/6 mouse were divided into 4 groups as 1 month,6 months,10 months and 19 months old and each group has 6 cases.Thymus tissue was removed and thymic stromal cells were isolated.And thymus epithelial cells were washed out by CD45 antibody and LS column after anesthesia.Fluorescence quantitative real-time PCR and Western blot were used to detect the changes of miR-194 and PTPN12 gene expression in thymus epithelial cells with aging.miR-194 and PTPN12 luciferase reporter vectors were transfected into HEK293 cells,and the auto fluorescence values were detected at 24 h and 48 h,respectively in vitro.Results:The expression level of miR-194 decreased(P<0.05),while the expression level of PTPN12 mRNA increased(P<0.05)as the age increased.And the correlation between miR-194 and PTPN12 mRNA expression was found to be negative(P<0.05).In vitro,luciferase reporter gene results show that miR-194 has a direct effect on the 3'UTR region of PTPN12 gene and had the highest binding efficiency in 48 h.Conclusion:PTPN12 is one of the target genes of miR-194,which is involved in the aging process of thymus and is an important factor regulating the function of thymic epithelial cells.

关 键 词：胸腺 增龄性萎缩 miR-194 PTPN12 

分 类 号：R392.2[医药卫生—免疫学] R394.4[医药卫生—基础医学]