缺氧诱导因子2α对含CUB结构域蛋白1的调控及在肝癌转移中的作用  

Effect of HIF-2α on regulating CDCP1 to promote hepatocellular carcinoma metastasis

作　　者：曹曼卿 高君蓉 黄佳飞 游阿彬 郭治贵 周洪渊[1] 房峰[1] 张伟 朱晓琳[1] 张倜[1] 

机构地区：[1]天津医科大学肿瘤医院肝胆科国家肿瘤临床医学研究中心天津市肿瘤防治重点实验室,300060

出　　处：《中华肿瘤杂志》2017年第1期18-23,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金（81372635、81101871、81201644）;天津市自然科学基金（11JCZDJC18800）

摘　　要：目的探讨缺氧诱导因子2α（HIF-2α）对CUB结构域蛋白1（CDCP1）的调控及其在肝癌转移中的作用。方法采用小干扰RNA（siRNA）和慢病毒转染的方法敲减和稳定过表达HIF-2α的肝癌细胞系MHCC97H，采用Western blot和实时荧光定量PCR检测CDCP1mRNA和蛋白的表达，采用Transwell侵袭实验检测HIF-2α对细胞侵袭能力的影响，采用免疫组化染色方法检测肝癌组织标本中CDCP1的表达。结果HIF-2α和CDCP1均能在乏氧条件下被诱导，且CDCP1在乏氧条件下的活化依赖于HIF-2α的表达。在肝癌细胞中，CDCP1在转录和翻译水平均受到HIF-2α的调控，过表达HIF-2α后，CDCP1mRNA的表达水平为5．92±0．28，高于阴性对照组（1．00±0．12），差异有统计学意义（P〈0．05）：敲减HIF-2α 48h和72h后，MHCC97H细胞中CDCP1mRNA的表达水平分别为0．25±0．04和0．18±0．02，均低于对照组（分别为1．00±0．18和1．00±0．19），差异均有统计学意义（均P〈0．05）。CDCP1的表达与患者的无瘤生存有关（P〈0．05）。结论CDCP1的表达与肝癌的进展有关，CDCP1的表达受HIF-2α的调控。通过靶向抑制HIF-2α和（或）CDCP1的表达可能对肝癌转移起到一定的抑制作用。Objective To investigate the effect of hypoxia inducible factor 2α （HIF-2α） on regulating CUB domain-containing protein 1 （ CDCP1 ） and its role in hepatocellular carcinoma metastasis. Methods HIF-2α-knocked down and HIF-2α-stably overexpressing cells （MHCC97H） were prepared by small interfering RNA （siRNA） and lentivirus transfection, respectively. The expression of CDCP1 protein and mRNA in the above cells was detected by western blot and real-time PCR. The effect of HIF-2α on cell invasion ability was determined by Transwell assay. Furthermore, immunohistochemical staining was performed to detect the expression of CDCP1 in human HCC tissue samples. Results Both HIF-2α and CDCPI were induced under bypoxic conditions. The activation of CDCP1 under hypoxic conditions was dependent on the expression of HIF-2α.When HIF-2α was overexpressed, the mRNA level of CDCP1 was greatly upregulated （5.92±0.28, P〈0.05）. When HIF-2α was knocked down by siRNA for 48 h and 72 h, the expression of CDCP1 was significantly downregulated （48 h： 0.25±0.04; 72 h： 0.18±0.02, all P〈 0.05 ）. Moreover, analysis of human HCC samples showed that CDCP1 expression was correlated with tumor- free survival （P〈0.05）. Conclusions The results of this study indicate that the expression of CDCP1 is regulated by HIF-2α and is correlated with the progression of HCC. Inhibition of HIF-2α/CDCP1 may play certain inhibitory role in the metastasis of HCC.

关 键 词：癌 肝细胞 肿瘤转移 缺氧诱导因子2α CUB结构域蛋白1 

分 类 号：R735.7[医药卫生—肿瘤]