CREB3 facilitates Donafenib resistance in hepatocellular carcinoma cells via the LSD1/CoREST/p65 axis by transcriptionally activating long noncoding RNA ZFAS1

IF 5 2区医学 Q2 Medicine Translational Oncology Pub Date : 2023-11-29 DOI:10.1016/j.tranon.2023.101684

Xunbo Hou , Qiannan Xu , Ruibao Liu

{"title":"CREB3 facilitates Donafenib resistance in hepatocellular carcinoma cells via the LSD1/CoREST/p65 axis by transcriptionally activating long noncoding RNA ZFAS1","authors":"Xunbo Hou , Qiannan Xu , Ruibao Liu","doi":"10.1016/j.tranon.2023.101684","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>Drug resistance greatly limits the therapeutic effect of a drug. This study aimed to explore the role of long noncoding RNA ZFAS1 in Donafenib resistance of hepatocellular carcinoma (HCC) cells.</p></div><div><h3>Methods</h3><p>The expression of CREB3, ZFAS1, and p65 in HCC cell lines was measured by RT-qPCR and western blotting. After transfection with sh-ZFAS1, sh-CREB3, or sh-CREB3 + oe-p65 in Donafenib-resistent (DR) HCC cell lines, the transfection efficiency was evaluated by RT-qPCR and western blotting. The proliferation and IC<sub>50</sub> to Donafenib of HCC cell lines was examined by MTT assay. Cell proliferation and apoptosis were examined by colony formation and flow cytometry assays. Then, the correlation amongst CREB3, ZFAS1, LSD1/CoREST, and p65 was analysed by ChIP, dual-luciferase reporter gene, and RIP assays.</p></div><div><h3>Results</h3><p>ZFAS1, CREB3, and p65 were upregulated in HepG2-DR and Huh7-DR cells. Silencing of ZFAS1 or CREB3 enhanced the sensitivity of HCC cells to Donafenib, inhibited cell proliferation and IC<sub>50</sub>, and increased cell apoptosis, which were reversed by p65 overexpression. Mechanistically, CREB3 bound to ZFAS1 promoter to augment ZFAS1 transcriptional expression, and ZFAS1 recruited LSD1/CoREST to the p65 promoter region to decrease H3K4 methylation and elevate p65 transcriptional expression.</p></div><div><h3>Conclusion</h3><p>CREB3 overexpression contributed to Donafenib resistance in HCC cells by activating the ZFAS1/p65 axis.</p></div>","PeriodicalId":48975,"journal":{"name":"Translational Oncology","volume":"44 ","pages":"Article 101684"},"PeriodicalIF":5.0000,"publicationDate":"2023-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1936523323000700/pdfft?md5=8829a80b83c6fc535ab0335ede8b9412&pid=1-s2.0-S1936523323000700-main.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Translational Oncology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1936523323000700","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Medicine","Score":null,"Total":0}

引用次数: 0

Abstract

Objective

Drug resistance greatly limits the therapeutic effect of a drug. This study aimed to explore the role of long noncoding RNA ZFAS1 in Donafenib resistance of hepatocellular carcinoma (HCC) cells.

Methods

The expression of CREB3, ZFAS1, and p65 in HCC cell lines was measured by RT-qPCR and western blotting. After transfection with sh-ZFAS1, sh-CREB3, or sh-CREB3 + oe-p65 in Donafenib-resistent (DR) HCC cell lines, the transfection efficiency was evaluated by RT-qPCR and western blotting. The proliferation and IC₅₀ to Donafenib of HCC cell lines was examined by MTT assay. Cell proliferation and apoptosis were examined by colony formation and flow cytometry assays. Then, the correlation amongst CREB3, ZFAS1, LSD1/CoREST, and p65 was analysed by ChIP, dual-luciferase reporter gene, and RIP assays.

Results

ZFAS1, CREB3, and p65 were upregulated in HepG2-DR and Huh7-DR cells. Silencing of ZFAS1 or CREB3 enhanced the sensitivity of HCC cells to Donafenib, inhibited cell proliferation and IC₅₀, and increased cell apoptosis, which were reversed by p65 overexpression. Mechanistically, CREB3 bound to ZFAS1 promoter to augment ZFAS1 transcriptional expression, and ZFAS1 recruited LSD1/CoREST to the p65 promoter region to decrease H3K4 methylation and elevate p65 transcriptional expression.

Conclusion

CREB3 overexpression contributed to Donafenib resistance in HCC cells by activating the ZFAS1/p65 axis.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

CREB3通过转录激活长链非编码RNA ZFAS1，通过LSD1/CoREST/p65轴促进肝癌细胞多纳非尼耐药

目的:耐药严重限制了药物的治疗效果。本研究旨在探讨长链非编码RNA ZFAS1在肝细胞癌(HCC)细胞多纳非尼耐药中的作用。方法采用RT-qPCR和western blotting检测肝癌细胞中CREB3、ZFAS1、p65的表达。将sh-ZFAS1、sh-CREB3或sh-CREB3 + e-p65转染多纳非尼耐药(DR) HCC细胞系后，采用RT-qPCR和western blotting评价转染效率。MTT法检测肝癌细胞株的增殖及对多那非尼的IC50。采用集落形成和流式细胞术检测细胞增殖和凋亡。然后，通过ChIP、双荧光素酶报告基因和RIP分析CREB3、ZFAS1、LSD1/CoREST和p65之间的相关性。结果HepG2-DR和Huh7-DR细胞中zfas1、CREB3和p65表达上调。沉默ZFAS1或CREB3增强HCC细胞对Donafenib的敏感性，抑制细胞增殖和IC50，增加细胞凋亡，p65过表达逆转。机制上，CREB3结合ZFAS1启动子增强ZFAS1转录表达，ZFAS1将LSD1/CoREST募集到p65启动子区域，降低H3K4甲基化，提高p65转录表达。结论creb3过表达通过激活ZFAS1/p65轴参与HCC细胞多纳非尼耐药。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Translational Oncology ONCOLOGY-

CiteScore

8.40

自引率

2.00%

发文量

314

审稿时长

54 days

期刊介绍： Translational Oncology publishes the results of novel research investigations which bridge the laboratory and clinical settings including risk assessment, cellular and molecular characterization, prevention, detection, diagnosis and treatment of human cancers with the overall goal of improving the clinical care of oncology patients. Translational Oncology will publish laboratory studies of novel therapeutic interventions as well as clinical trials which evaluate new treatment paradigms for cancer. Peer reviewed manuscript types include Original Reports, Reviews and Editorials.