Ye Jia-Zhou, Yan Shu-Mei, Yuan Chun-Ling, Wu Hui-Ni, Zhang Jin-Yan, Liu Zhi-Hui, Li Yong-Qiang, Luo Xiao-Ling, Lin Yan, Liang Rong
Department of Hepatobiliary Surgery, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, P. R. China.
Department of Pathology, Sun Yat-Sen University Cancer Center; State Key Laboratory of Oncology in South China; Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, P. R. China.
J Cancer. 2018 Jan 1;9(2):415-423. doi: 10.7150/jca.19185. eCollection 2018.
GP73 is a new hepatocellular carcinoma (HCC) marker, which is highly expressed in hepatocellular carcinoma and closely relates to prognosis. This study was to investigate the effects of GP73 on cellular proliferation, apoptosis, oxaliplatin (OXA) resistance and secretory clusterin (sCLU) of HCC cells. Western blot and immunofluorescence was used to detect the expression of GP73 in 8 types of commonly used HCC cell lines. Drug resistance was induced by increasing concentration gradient method. The drug-resistant human HCC cell lines underwent GP73 overexpression or inhibition. Flow cytometry were used to detect the proliferation and apoptosis of HCC cell lines. The changes of sCLU were detected by enzyme-linked immunosorbent assay (ELISA). The expression of GP73 in MHC-97H cells was the highest and in Hep3B cells the lowest. The expression of GP73 was found further elevated in OXA-resistant MHC-97H cells. After the knockdown of GP73 in OXA-resistant 97H cells, the IC of OXA decreased and the ability of cell proliferation decreased significantly. After over-expression of GP73 in OXA-resistant Hep3B cells, the IC of OXA increased and the cell proliferation ability increased, showing that GP73 is critical for OXA resistant in HCC cell lines; No significant change of sCLU level in GP73 overexpressed Hep3B and GP73 blocked MHCC-97H were identified. The expression level of GP73 is critical for the resistance of OXA in HCC cell lines.
GP73是一种新的肝细胞癌（HCC）标志物，在肝细胞癌中高表达且与预后密切相关。本研究旨在探讨GP73对肝癌细胞的细胞增殖、凋亡、奥沙利铂（OXA）耐药性及分泌性簇集素（sCLU）的影响。采用蛋白质免疫印迹法和免疫荧光法检测8种常用肝癌细胞系中GP73的表达。通过浓度梯度递增法诱导耐药。对耐药的人肝癌细胞系进行GP73过表达或抑制处理。采用流式细胞术检测肝癌细胞系的增殖和凋亡情况。通过酶联免疫吸附测定（ELISA）检测sCLU的变化。GP73在MHC-97H细胞中的表达最高，在Hep3B细胞中最低。在奥沙利铂耐药的MHC-97H细胞中发现GP73的表达进一步升高。在奥沙利铂耐药的97H细胞中敲低GP73后，奥沙利铂的半数抑制浓度（IC）降低，细胞增殖能力显著下降。在奥沙利铂耐药的Hep3B细胞中过表达GP73后，奥沙利铂的IC增加，细胞增殖能力增强，表明GP73对肝癌细胞系的奥沙利铂耐药性至关重要；在GP73过表达的Hep3B细胞和GP73阻断的MHCC-97H细胞中，未发现sCLU水平有显著变化。GP73的表达水平对肝癌细胞系的奥沙利铂耐药性至关重要。
