KRT23在肝细胞癌中的表达及对细胞生物学的影响

Expression of KRT23 in hepatocellular carcinoma and its impacts on cell biology

目的探讨角蛋白23(Keratin 23,KRT23)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及其对细胞增殖、迁移和侵袭的影响。方法应用免疫组织化学法检测88例HCC患者癌组织和癌旁正常组织中KRT23蛋白表达,并分析KRT23蛋白表达水平与HCC患者临床病理特征的相关性。体外培养人正常肝细胞系(L02)和HCC细胞系(Huh-7、HepG2、Bel-7402和MHCC97-H),qRT-PCR和Western blotting检测细胞中KRT23 mRNA和蛋白表达。取MHCC97-H细胞和HepG2细胞,构建KRT23敲低/过表达细胞系。CCK-8法检测细胞增殖活力,Transwell实验检测细胞迁移和侵袭能力,Western blotting检测细胞中EMT相关蛋白E-cadherin、N-cadherin、Vimentin表达。结果在HCC组织/细胞中KRT23蛋白均呈高表达,且KRT23蛋白表达水平与HCC的肿瘤大小、TNM分期和淋巴结转移有关(P均<0.05)。敲低KRT23可降低HCC细胞增殖活力,抑制细胞迁移、侵袭和N-cadherin、Vimentin表达,升高E-cadherin表达(P均<0.05);KRT23过表达可促进HCC细胞增殖、迁移和侵袭,降低E-cadherin表达,升高N-cadherin、Vimentin表达(P均<0.05)。结论KRT23在HCC中高表达,敲低KRT23可能通过抑制EMT,抑制HCC细胞增殖、迁移和侵袭。

Objective To investigate the expression of Keratin 23(KRT23)in hepatocellular carcinoma(HCC)and its impacts on cell proliferation,migration and invasion.Methods Immunohistochemistry was used to detect the expression of KRT23 protein in HCC tissue and adjacent normal tissue of 88 HCC patients,and the correlation between the expression level of KRT23 protein and the clinicopathological characteristics of HCC patients was analyzed.Human normal liver cell lines(L02)and HCC cell lines(Huh-7,HepG2,Bel-7402 and MHCC97-H)were cultured in vitro,and the mRNA and protein expressions of KRT23 in cells were detected by qRT-PCR and Western blotting.MHCC97-H cells and HepG2 cells were selected to construct KRT23 knockdown/overexpressing cell lines.The cell proliferation activity was detected by CCK-8 method,Transwell assay was used to detect cell migration and invasion abilities,the expression of EMT-related proteins(E-cadherin,N-cadherin,Vimentin)in the cells was detected by Western blotting.Results KRT23 protein was highly expressed in HCC tissues/cells,and the expression level of KRT23 protein was correlated with tumor size,TNM stage and lymph node metastasis of HCC(all P<0.05).Knockdown of KRT23 could reduce the proliferation of HCC cells,inhibit cell migration,invasion and the expression of N-cadherin and Vimentin,and increase the expression of E-cadherin(all P<0.05);overexpression of KRT23 could promote the proliferation,migration and invasion of HCC cells,decrease the expression of E-cadherin,and increase the expressions of N-cadherin and Vimentin(all P<0.05).Conclusion KRT23 is highly expressed in HCC,and knockdown of KRT23 may inhibit the proliferation,migration and invasion of HCC cells by inhibiting EMT.