Cao M Q, Gao J R, Huang J F, You A B, Guo Z G, Zhou H Y, Fang F, Zhang W, Zhu X L, Zhang T
Department of Hepatobiliary Surgery, Cancer Institute and Hospital, Tianjin Medical University, Key Laboratory of Cancer Prevention and Therapy, National Clinical Research Center for Cancer, Tianjin 300060, China.
Zhonghua Zhong Liu Za Zhi. 2017 Jan 23;39(1):18-23. doi: 10.3760/cma.j.issn.0253-3766.2017.01.004.
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. 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. Both HIF-2α and CDCP1 were induced under hypoxic 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, <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 <0.05). Moreover, analysis of human HCC samples showed that CDCP1 expression was correlated with tumor-free survival (<0.05). 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.
探讨缺氧诱导因子2α（HIF-2α）对含CUB结构域蛋白1（CDCP1）的调控作用及其在肝细胞癌转移中的作用。分别采用小干扰RNA（siRNA）和慢病毒转染制备HIF-2α敲低和HIF-2α稳定过表达的细胞（MHCC97H）。通过蛋白质免疫印迹法和实时荧光定量PCR检测上述细胞中CDCP1蛋白和mRNA的表达。采用Transwell实验检测HIF-2α对细胞侵袭能力的影响。此外，通过免疫组织化学染色检测人肝癌组织样本中CDCP1的表达。缺氧条件下HIF-2α和CDCP1均被诱导。缺氧条件下CDCP1的激活依赖于HIF-2α的表达。当HIF-2α过表达时，CDCP1的mRNA水平显著上调（5.92±0.28，P<0.05）。当用siRNA敲低HIF-2α 48小时和72小时时，CDCP1的表达显著下调（48小时：0.25±0.04；72小时：0.18±0.02，均P<0.05）。此外，对人肝癌样本的分析表明，CDCP1的表达与无瘤生存期相关（P<0.05）。本研究结果表明，CDCP1的表达受HIF-2α调控，且与肝癌进展相关。抑制HIF-2α/CDCP1可能对肝癌转移起一定的抑制作用。
