Metastin Inhibits Migration and Invasion of Renal Cell Carcinoma with Overexpression of Metastin Receptor
Metastin Inhibits Migration and Invasion of Renal Cell Carcinoma with Overexpression of Metastin Receptor
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DOI：
10.1016/j.eururo.2008.02.048
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发表时间：
2009-02-01
期刊：
影响因子：
23.4
通讯作者：
Terachi, Toshiro
中科院分区：
文献类型：
--
作者：
Shoji, Sunao;Tang, Xian Yan;Terachi, Toshiro
背景：KiSS-1基因的终端肽Metastin被认为具有抑制细胞活性的作用。目的：研究肾癌组织是否表达Metastin或其受体，阐明Metastin能否抑制肾癌细胞在体外的迁移和/或侵袭和/或增殖。设计、地点和对象：25例肾细胞癌标本。新鲜肾癌组织制备实时定量RT-PCR，石蜡包埋和福尔马林固定的组织块进行免疫组织化学检测。肾癌细胞株CAKY-1和ACHN用于细胞迁移、侵袭和增殖检测。免疫组织化学采用EnVision系统。用创伤愈合实验和Matrigel实验检测肾癌细胞系的迁移和侵袭能力。细胞计数试剂盒检测细胞增殖情况。在Meta系统下观察细胞形态。结果与局限性：实时定量RT-PCR25例肾癌组织中Metastin受体的表达水平显著高于非肿瘤性肾皮质组织(p=0.011)。免疫组织化学检测所有肾细胞癌组织中均有Metastin受体蛋白表达。体外实验表明，Metastin可抑制Caki-1和ACHN细胞的迁移和侵袭。相反，它对细胞增殖没有影响。胃动素(10MU/L)诱导CAKI-1和ACHN细胞过度粘连和应力纤维的形成，这种现象可被药物Rho-Kinase抑制剂Y-27632抑制。我们证明，Metastin可以抑制肾癌细胞系的迁移和侵袭，这是由Rho-Kinase抑制剂调节的。因此，Metastin及其受体可能是抑制肾细胞癌的靶点。(C)2008年欧洲泌尿外科协会。爱思唯尔出版，版权所有。
Background: Metastin, the final peptide of the KiSS-1 gene, has been proposed to suppress cell motility.Objective: This study investigated whether renal cell carcinoma (RCC) tissue expresses metastin or its receptor, and clarified whether metastin can suppress migration and/or invasion and/or proliferation of RCC cells in vitro.Design, Setting, and Participants: Twenty-five RCC samples were submitted. Fresh RCC tissues were prepared for real-time RT-PCR, and formalin-fixed and paraffin-embedded tissues blocks were examined by immunohistochemistry. RCC cell lines Caki-1 and ACHN were supplied for cell migration, invasion, and proliferation assays.Measurements: Real-time RT-PCR was performed by using Taq Man gene expression system. ENVISION system was used in immunohistochemistry. Wound-healing assay and matrigel assays were used to identify migration and invasion abilities of RCC cell lines. Cell Counting Kit-8 was applied to measure the cell proliferation. Cell morphology was examined under a META system. Statistical analysis was performed with SPSS(R)15.0J.Results and Limitations: In twenty-five RCC samples, the mRNA level of metastin receptor was identified to be significantly higher than non-neoplastic renal cortex by real-time RT-PCR (p = 0.011). Immunohistochemical study also detected metastin receptor protein in all RCC tumors. In vitro, this study showed that metastin inhibited migration and invasion of Caki-1 and ACHN cells. In contrast, it had no effects on cell proliferation. Metastin (10 mu mol/l) induced excessive formation of focal adhesions and stress fibers in Caki-1 and ACHN cells; this phenomenon was inhibited by pretreating pharmacological Rho-kinase inhibitor (Y-27632) to those cells.Conclusion: This is the first report regarding overexpression of the metastin receptor hOT7T175 in human RCC. We demonstrate that metastin can inhibit migration and invasion of the RCC cell line, which is regulated by a Rho-kinase inhibitor. Metastin and its receptor are therefore probable targets for suppressing RCC. (C) 2008 European Association of Urology. Published by Elsevier B.V. All rights reserved.