当前位置: X-MOL 学术Autoimmunity › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
miR-93-5p knockdown repressed hepatocellular carcinoma progression via increasing ERBB4 and TETs-dependent DNA demethylation
Autoimmunity ( IF 3.5 ) Pub Date : 2021-08-26 , DOI: 10.1080/08916934.2021.1969552
Yuqiang Li 1 , Bin Wu 1 , Rongli Sun 1 , Mingzhou Zhao 1 , Nan Li 2
Affiliation  

Abstract

Background

microRNAs (miRNAs) are involved in hepatocellular carcinoma (HCC) development and can control gene expression via directly targeting or regulating DNA methylation. This research aims to analyse the mechanism of miR-93-5p on HCC progression.

Methods

miR-93-5p, Erb-B2 receptor tyrosine kinase 4 (ERBB4) and ten-eleven translocation methyl-cytosine dioxygenases (TET1, TET2 and TET3) abundances were measured via quantitative reverse transcription polymerase chain reaction and Western blotting. The binding interaction was examined by dual-luciferase reporter analysis and chromatin immunoprecipitation. Cell proliferation and apoptosis were assessed via Cell Counting Kit-8, colony formation and flow cytometry. The DNA methylation of ERBB4 was detected via specific polymerase chain reaction. SNU-449 cells were subcutaneously inoculated into the BALB/c nude mice to establish the in vivo model for HCC, and the in vivo function of miR-93-5p was analysed by intratumoral injections of miR-93-5p antogomir.

Results

miR-93-5p abundance was enhanced and ERBB4 level was reduced in HCC tumour tissues of 62 patients and HCC cell lines, in contrast with that in paired normal tissues of 62 patients and normal cell lines. ERBB4 was targeted by miR-93-5p. miR-93-5p knockdown or ERBB4 overexpression repressed HCC cell proliferation and promoted apoptosis via decreasing cell viability and colony ability and inducing cycle arrest. ERBB4 silence attenuated the influence of miR-93-5p knockdown on cell proliferation and apoptosis. ERBB4 promoter DNA methylation level was enhanced in HCC samples and cell lines, and ERBB4 abundance was increased via TETs (TET1, TET2 and TET3). miR-93-5p targeted TETs to modulate ERBB4 abundance. TETs silence relieved the influence of miR-93-5p knockdown on cell proliferation and apoptosis. miR-93-5p knockdown decreased HCC growth in a xenograft model.

Conclusion

miR-93-5p knockdown repressed the progression of HCC via increasing ERBB4 and TETs-dependent DNA demethylation.



中文翻译:

miR-93-5p 敲低通过增加 ERBB4 和 TETs 依赖性 DNA 去甲基化抑制肝细胞癌进展

摘要

背景

microRNAs (miRNAs) 参与肝细胞癌 (HCC) 的发展,可以通过直接靶向或调节 DNA 甲基化来控制基因表达。本研究旨在分析miR-93-5p对HCC进展的作用机制。

方法

通过定量逆转录聚合酶链反应和蛋白质印迹测量 miR-93-5p、Erb-B2 受体酪氨酸激酶 4 (ERBB4) 和 11-11 易位甲基胞嘧啶双加氧酶 (TET1、TET2 和 TET3) 丰度。通过双荧光素酶报告基因分析和染色质免疫沉淀检查结合相互作用。通过Cell Counting Kit-8、集落形成和流式细胞术评估细胞增殖和凋亡。ERBB4 的 DNA 甲基化通过特异性聚合酶链反应检测。将 SNU-449 细胞皮下接种于 BALB/c 裸鼠体内,建立HCC体内模型通过瘤内注射miR-93-5p antogomir来分析miR-93-5p的功能。

结果

与 62 例患者的成对正常组织和正常细胞系相比,62 例患者的 HCC 肿瘤组织和 HCC 细胞系中 miR-93-5p 丰度增加,ERBB4 水平降低。ERBB4 被 miR-93-5p 靶向。miR-93-5p 敲低或 ERBB4 过表达通过降低细胞活力和集落能力并诱导细胞周期停滞来抑制 HCC 细胞增殖并促进细胞凋亡。ERBB4 沉默减弱了 miR-93-5p 敲低对细胞增殖和凋亡的影响。ERBB4 启动子 DNA 甲基化水平在 HCC 样品和细胞系中增强,ERBB4 丰度通过TET(TET1、TET2 和 TET3)。miR-93-5p 靶向 TET 以调节 ERBB4 丰度。TETs 沉默减轻了 miR-93-5p 敲低对细胞增殖和凋亡的影响。miR-93-5p 敲低降低了异种移植模型中 HCC 的生长。

结论

miR-93-5p 敲低通过增加 ERBB4 和 TETs 依赖性 DNA 去甲基化来抑制 HCC 的进展。

更新日期:2021-08-26
down
wechat
bug