Effects of TET2-targeting miR-27b on oxidized low-density lipoprotein-induced inflammatory responses and apoptosis in endothelial cells

Q4 Immunology and Microbiology

中华微生物学和免疫学杂志 Pub Date : 2019-11-30 DOI:10.3760/CMA.J.ISSN.0254-5101.2019.11.008

J. Bao, Bei Sun, P. Ma, Hui Xu, Lili Ji

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引用次数: 0

Abstract

Objective To investigate the effects of miR-27b targeting ten-eleven translocation methylcytosine dioxygenase 2 (TET2) on oxidized low-density lipoprotein (ox-LDL) induced inflammatory responses and apoptosis in endothelial cells. Methods Double luciferase reporter gene analysis verified the targeting effect of miR-27b on TET2. Human umbilical vein endothelial cells (HUVECs) were induced by ox-LDL in vitro. Eight groups were set up including control group, ox-LDL group, ox-LDL+ anti-miR-con group, ox-LDL+ anti-miR-27b group, ox-LDL+ pcDNA group, ox-LDL+ pcDNA-TET2 group, anti-miR-27b+ si-con group and anti-miR-27b+ si-TET2 group. qRT-PCR was used to detect the expression of miR-27b and TET2 at mRNA level. Cell viability was detected by MTT assay. Cell apoptosis rate was detected by flow cytometry. Western blot was used to detect the expression of TET2, Cyclin D1 and caspase-3 at protein level. Results TET2 was the target gene of miR-27b. TET2 expression could be negatively regulated by miR-27b. ox-LDL increased the expression of miR-27b and reduced the expression of TET2 in HUVECs. The secretion of inflammatory factors and apoptosis rates of HUVECs in the control, ox-LDL+ anti-miR-27b, ox-LDL+ pcDNA-TET2 and anti-miR-27b+ si-con groups were significantly lower than those in the ox-LDL, ox-LDL+ anti-miR-con, ox-LDL+ pcDNA and anti-miR-27b+ si-TET2 groups, respectively (P<0.05). Conclusions miR-27b promoted ox-LDL-induced inflammatory responses and apoptosis in endothelial cells through down-regulating the expression of TET2. Key words: miR-27b; ox-LDL; TET2; HUVEC; Inflammatory response; Apoptosis

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靶向miR-27b的TET2对氧化低密度脂蛋白诱导的内皮细胞炎症反应和凋亡的影响

目的研究miR-27b靶向十-十一易位甲基胞嘧啶双加氧酶2（TET2）对氧化低密度脂蛋白（ox-LDL）诱导的内皮细胞炎症反应和凋亡的影响。方法双荧光素酶报告基因分析验证了miR-27b对TET2的靶向作用。ox-LDL体外诱导人脐静脉内皮细胞（HUVECs）。设8组，分别为对照组、ox-LDL组、ox-DLDL+抗-miR-con组、ox-RLDL+反-miR-27b组、ox-lLDL+pcDNA组、ox/LLDL+pcDNA-TET2组、抗-miR-27b+si-con组和抗-miR27b+si-T2组。qRT-PCR用于检测miR-27b和TET2在mRNA水平上的表达。MTT法检测细胞活力。流式细胞仪检测细胞凋亡率。蛋白质印迹法检测TET2、Cyclin D1和胱天蛋白酶-3在蛋白水平的表达。结果TET2是miR-27b的靶基因。TET2的表达可能受到miR-27b的负调控。ox-LDL增加了HUVECs中miR-27b的表达并降低了TET2的表达。对照组、ox-LDL+抗-miR-27b、ox-LLD+pcDNA-TET2和抗-miR27b+si-con组的HUVECs的炎症因子分泌和凋亡率显著低于ox-LDL、ox-LDL+抗-miR con、ox-LD+pcDNA和抗-miR-27b+si-T2组，结论miR-27b通过下调TET2的表达，促进ox-LDL诱导的内皮细胞炎症反应和凋亡。关键词：miR-27b；ox-LDL；TET2；HUVEC；炎症反应；细胞凋亡

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来源期刊

中华微生物学和免疫学杂志 Immunology and Microbiology-Virology

CiteScore

0.50

自引率

0.00%

发文量

6906

期刊介绍： Chinese Journal of Microbiology and Immunology established in 1981. It is one of the series of journal sponsored by Chinese Medical Association. The aim of this journal is to spread and exchange the scientific achievements and practical experience in order to promote the development of medical microbiology and immunology. Its main contents comprise academic thesis, brief reports, reviews, summaries, news of meetings, book reviews and trends of home and abroad in this field. The distinguishing feature of the journal is to give the priority to the reports on the research of basic theory, and take account of the reports on clinical and practical skills.