Mechanism of citron rho-interacting serine/threonine kinase gene regulating adriamycin resistance in hepatocellular carcinoma cell line SK-Hep1

GUO Li

doi:10.16098/j.issn.0529-1356.2020.05.022

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (5) : 772-777. DOI: 10.16098/j.issn.0529-1356.2020.05.022

Mechanism of citron rho-interacting serine/threonine kinase gene regulating adriamycin resistance in hepatocellular carcinoma cell line SK-Hep1

GUO Li¹ ZHAO Hong² ZHANG Jun-tao² LIU Zhi-zhen² GONG Tao² YU Bao-feng^2*

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Abstract

Objective To investigate the regulatory effect of citron rho-interacting serine/threonine kinase (CIT) on drug resistance of adriamycin (ADM) resistant hepatocellular carcinoma cells and its mechanism. Methods Random nonsense negative sequence (si-NC), CIT small interfering RNA (si-CIT) trasfected into SK-Hep1/ADM cells with liposomes. Real-time PCR, Western blotting were used to detect the expression of CIT, X linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma protein 2 (Bcl-2), Bcl-2 related X gene (Bax), cytochrome C (Cyt C), phosphatase gene induction putative kinase 1 (PINK1), autosomal recessive adolescent Parkinson’s disease pathogenic gene (Parkin) and autophagy microtubule-associated protein light chain 3 antibody Ⅰ/Ⅱ(LC3-Ⅰ/Ⅱ); Cell counting kit 8(CCK-8), flow cytometry, and fluorescence probe(JC-10) staining were used to detect cell inhibition rate, apoptosis and mitochondrial membrane potential. Results The expression level of CIT in drug-resistant cells was significantly higher than that in normal cells. Knocking down CIT significantly increased the sensitivity to ADM. Knocking down CIT promoted drug-resistant cell apoptosis and mitochondrial autophagy, down-regulated cell mitochondrial membrane potential, and up-regulated Bax, Cyt C, and PINK1, Parkin,LC3-Ⅰ/Ⅱ, down-regulate XIAP, Bcl-2, Cyt C. Conclusion CIT gene could regulate the drug resistance of adriamycin  resistant HCC cells through mitochondrial pathway apoptosis and autophagy.

Key words

Citron rho-interacting serine/threonine kinase / Adriamycin-resistant hepatic carcinoma cells / Mitochondria / Autophagy / Western blotting

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GUO Li ZHAO Hong ZHANG Jun-tao LIU Zhi-zhen GONG Tao YU Bao-feng. Mechanism of citron rho-interacting serine/threonine kinase gene regulating adriamycin resistance in hepatocellular carcinoma cell line SK-Hep1[J]. Acta Anatomica Sinica. 2020, 51(5): 772-777 https://doi.org/10.16098/j.issn.0529-1356.2020.05.022

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