LINC00467 promotes hepatocellular carcinoma resistance to sorafenib by regulating CDK1 and CDC45

doi:10.3877/cma.j.issn.2095-3232.2020.03.019

Abstract

Abstract:

Objective

To explore the mechanism of LINC00467 in regulating the resistance of hepatocellular carcinoma (HCC) cells to sorafenib.

Methods

GEO dataset GSE73571 was analyzed by weighted gene co-expression network analysis (WGCNA) to screen the genes related to sorafenib resistance. The functional genes related to LINC00467 and the downstream genes were confirmed by GO/KEGG enrichment analysis. The sorafenib-resistant cell line HepG2-R was constructed and normal HepG2 cells were assigned as HepG2-P. The cytotoxicity of sorafenib was determined by CCK-8 assay and the IC₅₀ value was calculated. The mRNA expression of LINC00467, cyclin-dependent kinase 1 (CDK1) and cell division cyclin 45 (CDC45) were detected by fluorescent quantitative RT-PCR. The mRNA relative expression of LINC00467, CDK1 and CDC45 between two groups were statistically compared by t test.

Results

WGCNA identified LINC00467 as the sorafenib resistance-related gene. GO/KEGG enrichment analysis confirmed CDK1 and CDC45 as the functional genes related to LINC00467 and the downstream genes. Fluorescent quantitative RT-PCR found that the relative expression level of LINC00467 mRNA in HepG2-R cells was 2.49±0.30, significantly higher than 1 in HepG2-P cells (t=4.91, P<0.05). The relative expression level of LINC00467 mRNA was 0.36±0.04 after siRNA interference, significantly lower than 1 before siRNA interference (t=-14.60, P<0.05). siRNA-LINC00467 transfection could reduced the IC₅₀ value of sorafenib-resistant cells from 14.03 μmol/L to 7.33 μmol/L. The expression levels of CDK1 and CDC45 were up-regulated in sorafenib-resistant HepG2-R cells, whereas it was significantly down-regulated when LINC00467 interfered by siRNA.

Conclusions

Based on the mining of public database, it is found that LINC00467 is highly expressed in HCC cells. LINC00467 participates in the regulation of cell cycle and DNA damage repair process probably by up-regulating the expression of CDK1 and CDC45, thereby promoting the resistance of HCC cells to sorafenib.

Key words: Carcinoma, hepatocellular, Sorafenib, Drug resistance, LINC00467, CDK1, CDC45

Zhirong Lin, Wenliang Tan, Shenghai Qin, Xinming Chen, Yajin Chen, Changzhen Shang. LINC00467 promotes hepatocellular carcinoma resistance to sorafenib by regulating CDK1 and CDC45[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2020, 09(03): 289-294.

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