Liu Yan, Qin Zhenbang, Yang Kuo, Liu Ranlu, Xu Yong
Prostate Disease Laboratory, Tianjin Institute of Urology, Tianjin 300211, P.R. China.
Oncol Rep. 2017 Mar;37(3):1521-1528. doi: 10.3892/or.2017.5378. Epub 2017 Jan 17.
The Cripto-1 (CR-1) derived EGF-CFC family was overexpressed in tumor development enhancing proliferation, epithelial-mesenchymal transition (EMT) and migration of tumor cells. However, correlation between CR-1 and prostate cancer (PCa) remains still unclear. In the present study, we proved that CR-1 was expressed in PCa and its function was in the progression of PCa. Compared with benign prostatic hyperplasia (BPH) tissues, we confirmed that PCa tissues had high expression of CR-1 by immunohistochemistry and statistical data showed that CR-1 promoted properties of EMT in PCa tissues, including the downregulation of the cell adhesion molecules β-catenin (membrane) and E-cadherin while upregulating transcription factors β-catenin. Overexpression of CR-1 had close relationship with PSA, Gleason, clinical staging and lymph node metastasis in PCa patients. Then, we found that PC-3 cells transfected with CR-1-shRNA inhibited EMT using RT-PCR, RT-qPCR, western blotting and immunofluorescence. Also, we evaluated cell invasive ability in vitro by transwell and wound-healing assay. Our data showed that transfected CR-1-shRNA altered EMT including β-catenin, E-cadherin, c-myc, GSK-3, p-GSK and Wnt/β-catenin pathway in PC-3. It also suppressed PC-3 cell migration. Additionally, our results displayed that Licl had antitumor activity against PC-3 through the inhibition of Wnt/β-catenin pathway. Inhibition of cell viability was dose-time dependent. The present study proved that CR-1 regulates EMT of PCa by Wnt/β-catenin pathway. Hence, CR-1 may provide a new biological marker, and possibly contributes to clinical treatment against PCa.
源自Cripto-1（CR-1）的表皮生长因子-CFC家族在肿瘤发生发展过程中过表达，可促进肿瘤细胞增殖、上皮-间质转化（EMT）及迁移。然而，CR-1与前列腺癌（PCa）之间的相关性仍不明确。在本研究中，我们证实CR-1在PCa中表达，且其功能与PCa进展相关。通过免疫组织化学，我们确认与良性前列腺增生（BPH）组织相比，PCa组织中CR-1高表达，统计数据表明CR-1促进PCa组织中的EMT特性，包括细胞黏附分子β-连环蛋白（膜）和E-钙黏蛋白下调，同时上调转录因子β-连环蛋白。CR-1过表达与PCa患者的前列腺特异性抗原（PSA）、 Gleason评分、临床分期及淋巴结转移密切相关。然后，我们发现用CR-1短发夹RNA（shRNA）转染的PC-3细胞通过逆转录-聚合酶链反应（RT-PCR）、实时定量RT-PCR（RT-qPCR）、蛋白质免疫印迹法和免疫荧光抑制EMT。此外，我们通过Transwell小室和伤口愈合试验评估细胞体外侵袭能力。我们的数据显示，转染CR-1-shRNA改变了PC-3细胞中的EMT，包括β-连环蛋白、E-钙黏蛋白、c-myc、糖原合成酶激酶-3（GSK-3）、磷酸化GSK（p-GSK）和Wnt/β-连环蛋白信号通路。它还抑制了PC-3细胞迁移。此外，我们的结果显示氯化锂（Licl）通过抑制Wnt/β-连环蛋白信号通路对PC-3具有抗肿瘤活性。细胞活力的抑制呈剂量-时间依赖性。本研究证明CR-1通过Wnt/β-连环蛋白信号通路调节PCa的EMT。因此，CR-1可能提供一种新的生物标志物，并可能有助于PCa的临床治疗。
