转录因子CREB调节小鼠CD4+淋巴细胞中miR-30a的表达

doi:10.16352/j.issn.1001-6325.2023.08.1229

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (8): 1229-1233.doi: 10.16352/j.issn.1001-6325.2023.08.1229

转录因子CREB调节小鼠CD4⁺淋巴细胞中miR-30a的表达

韩晶晶, 王萱若, 张欣怡, 高涵, 成秀梅, 杨留才, 曲学彬^*

江苏医药职业学院基础医学部,江苏盐城 224005

收稿日期:2022-11-29 修回日期:2023-02-20 出版日期:2023-08-05 发布日期:2023-07-26
通讯作者: ^*jaty2222@163.com
基金资助:
江苏省卫生健康委医学科研项目(Z2022018)

Transcription factor CREB regulates the expression of miR-30a in mouse CD4⁺ lymphocytes

HAN Jingjing, WANG Xuanruo, ZHANG Xinyi, GAO Han, CHENG Xiumei, YANG Liucai, QU Xuebin^*

Department of Basic Medicine, Jiangsu Vocational College of Medicine, Yancheng 224005, China

Received:2022-11-29 Revised:2023-02-20 Online:2023-08-05 Published:2023-07-26
Contact: ^*jaty2222@163.com

摘要/Abstract

摘要： 目的研究CD4⁺淋巴细胞系中转录因子cAMP反应元件结合蛋白(CREB)对miR-30a的表达调节作用。方法利用UCSC和Ensembl网站确定miR-30a基因启动子区;EMBOSS Cpgplot网站分析启动子区DNA序列的甲基化情况;ECR Browser网站分析启动子区转录因子结合区域及其保守性;JASPAR数据库筛选启动子区结合的转录因子;染色质免疫沉淀检测小鼠CD4⁺淋巴细胞中miR-30a基因启动子区CREB的结合量;依据培养基中是否添加CREB抑制剂KG-501将小鼠CD4⁺淋巴细胞分为对照组、溶剂对照组和KG-501处理组,RT-qPCR检测KG-501对细胞中miR-30a表达的影响。结果 miR-30a基因启动子区未发现可被甲基化的CpG岛;启动子区含有高保守性的转录因子结合序列,其中包含多个CREB的结合位点;转录因子CREB直接结合于miR-30a基因启动子区的上游区段;添加KG-501可明显下调细胞中miR-30a的表达(P＜0.05)。结论转录因子CREB直接结合于小鼠CD4⁺淋巴细胞中miR-30a基因启动子区的特定位点,调节miR-30a的表达。

关键词: 微小RNA, DNA甲基化, 转录因子, cAMP反应元件结合蛋白

Abstract: Objective To study the regulatory effect of the transcription factor cAMP response element binding protein (CREB) on the expression of miR-30a in CD4⁺ lymphocytes. Methods The promoter region of miR-30a gene was determined by UCSC and Ensembl website. The methylation of DNA sequence in the promoter region was calculated on EMBOSS Cpgplot website. The transcription factor binding region and its conservation were analyzed via ECR Browser website. The special binding transcription factors were predicted with support from JASPAR database. Chromatin immunoprecipitation assay was used to confirm the binding of CREB in the promoter region of miR-30a gene in mouse CD4⁺ lymphocytes. According to whether CREB inhibitor KG-501 was added to the culture medium, mouse CD4⁺ lymphocytes were divided into three groups, control, vehicle and KG-501 treated group. The effect of KG-501 on the expression of miR-30a in the cells was detected by RT-qPCR. Results None methylated CpG islands were found in the promoter region of miR-30a gene. The promoter region contained highly conserved transcription factor binding sequences, including several CREB binding sites. Transcription factor CREB bound to the upstream of miR-30a gene promoter region. KG-501 significantly down-regulated the expression of miR-30a(P＜0.05). Conclusions Transcription factor CREB binds to specific sites in the promoter region of miR-30a gene to regulate the expression of miR-30a in mouse CD4⁺ lymphocytes.

Key words: microRNA, DNA methylation, transcription factor, cAMP response element binding protein

中图分类号:

R392.11

韩晶晶, 王萱若, 张欣怡, 高涵, 成秀梅, 杨留才, 曲学彬. 转录因子CREB调节小鼠CD4⁺淋巴细胞中miR-30a的表达[J]. 基础医学与临床, 2023, 43(8): 1229-1233.

HAN Jingjing, WANG Xuanruo, ZHANG Xinyi, GAO Han, CHENG Xiumei, YANG Liucai, QU Xuebin. Transcription factor CREB regulates the expression of miR-30a in mouse CD4⁺ lymphocytes[J]. Basic & Clinical Medicine, 2023, 43(8): 1229-1233.

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转录因子CREB调节小鼠CD4⁺淋巴细胞中miR-30a的表达

Transcription factor CREB regulates the expression of miR-30a in mouse CD4⁺ lymphocytes

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转录因子CREB调节小鼠CD4+淋巴细胞中miR-30a的表达

Transcription factor CREB regulates the expression of miR-30a in mouse CD4+ lymphocytes

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转录因子CREB调节小鼠CD4⁺淋巴细胞中miR-30a的表达

Transcription factor CREB regulates the expression of miR-30a in mouse CD4⁺ lymphocytes