The effect of AHR small interference on hepatoma HCCLM3 cell proliferation in vitro and tumor growth in vivo / 肿瘤

ABSTRACT

Objective: To investigate the effect of aryl hydrocarbon receptor (AHR) RNA interference on hepatoma HCCLM3 cell proliferation and migration and to explore its possible mechanism. Methods: After HCCLM3 cells transfection with specific AHR-small interference RNA (siRNA), the expression level of AHR mRNA was detected by real-time fluorescence quantitative-PCR, and the expression levels of AHR, stress-activated protein kinases (SAPK)/c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (ERK 1/2) and c-Jun phosphorylation were detected by Western blotting. Then the proliferation and migration of HCCLM3 cells after transfection with AHR-siRNA were detected by cell counting kit-8 (CCK-8) and Transwell assay, respectively. Synthesize AHR gene-specific small hairpin RNA (shRNA) to construct recombinant virus vector pMKO.1/puro-shAHR which stable knockdown AHR, and the recombinant vector was infected into HCCLM3 cells and then implanted into nude mice to construct subcutaneous tumor model. The growth of xenografted tumor in nude mice was examined. The expression levels of AHR protein in HCCLM3 cells and xenografted tumor tissues after infection with pMKO.1/puro-shAHR were detected by Western blotting. Results: The expression levels of AHR mRNA and protein were significantly reduced in HCCLM3 cells transfected with AHR-siRNA (P < 0.01), and the proliferation and migration of HCCLM3 cells were inhibited (P < 0.01). The expression levels of SAPK/JNK, ERK 1/2 and c-Jun phosphorylation were depressed (P < 0.01). The volume and weight of HCCLM3 cells xenografted tumor in nude mice after infection with pMKO.1/puro-shAHR were lower than those of the control group (HCCLM3 cells were infected with pMKO.1/puro-shNC) (P < 0.01). The expression levels of AHR protein in HCCLM3 cells and xenografted tissues after infection with pMKO.1/puro-shAHR were decreased (P < 0.01). Conclusion: AHR may promote HCCLM3 cell proliferation and migration in vitro by up-regulating the phosphorylation levels of ERK1/2, SAPK/JNK and c-Jun. Copyright © 2014 by TUMOR.