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

Xunbo Hou; Qiannan Xu; Ruibao Liu

doi:10.1016/j.tranon.2023.101684

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

Transl Oncol. 2024 Jun:44:101684. doi: 10.1016/j.tranon.2023.101684. Epub 2023 Nov 29.

Authors

Xunbo Hou¹, Qiannan Xu², Ruibao Liu³

Affiliations

¹ Department of Interventional, Harbin Medical University Cancer Hospital, No. 150, Haping Rd, Nangang District, Harbin, Heilongjiang 150081, PR China.
² Department of Anesthesiology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, PR China.
³ Department of Interventional, Harbin Medical University Cancer Hospital, No. 150, Haping Rd, Nangang District, Harbin, Heilongjiang 150081, PR China. Electronic address: xunbohou@hotmail.com.

PMID: 38641372
DOI: 10.1016/j.tranon.2023.101684

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.

Keywords: CREB3; Donafenib; Drug resistance; Hepatocellular carcinoma; LSD1/CoREST; ZFAS1; p65.