Decreased expression of REIC/Dkk-3 in human renal clear cell carcinoma
Decreased expression of REIC/Dkk-3 in human renal clear cell carcinoma
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DOI：
10.1097/01.ju.0000101047.64379.d4
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发表时间：
2004-03-01
影响因子：
6.6
通讯作者：
Huh, NH
中科院分区：
文献类型：
--
作者：
Kurose, K;Sakaguchi, M;Huh, NH
目的：检测可能的抑癌基因REIC/DKK-3在人肾透明细胞癌(RCCC)细胞系和散发性肾透明细胞癌手术标本中的表达。材料和方法：以人肾透明细胞癌细胞系(CAKI-1、CAKI-2、ACHN和KPK-1)和几个对照细胞系为对照，检测REIC/DKK-3基因的表达，并鉴定一种新的针对REIC/DKK-3蛋白的抗体。对20例肾癌患者的癌组织和癌旁组织进行分析。对其中17例和7例分别进行实时定量逆转录聚合酶链式反应、Western印迹分析和/或免疫组织化学分析。结果：证实了RCCC细胞株中ReIC/DKK-3mRNA和蛋白的表达降低，并证实了新抗体的特异性。在一项实时定量逆转录聚合酶链式反应研究中，17对肾癌组织及癌旁正常组织中REIC/DKK-3的mRNA表达水平在癌组织中显著降低(在15对中，下降了25%至约95%)。免疫组织化学和Western印迹分析显示，在所分析的14例肾癌中，REIC/DKK-3蛋白表达水平下降的有6例，分别为6例和13例。结论：REIC/DKK-3mRNA和蛋白水平的降低与肿瘤的分级和分期无关，提示REIC/DKK-3参与了肿瘤恶性转化的初始阶段。因此，REIC/DKK-3有可能成为治疗肾癌的新分子靶点。
Purpose: We examined the expression of REIC/Dkk-3, a possible candidate for a tumor suppressor gene, in human renal clear cell carcinoma (RCCC) cell lines and sporadic RCCC surgical specimens.Materials and Methods: Human RCCC cell lines (Caki-1, Caki-2, ACHN and KPK-1) and several control cell lines were used to examine the expression of REIC/Dkk-3 mRNA and characterize a newly raised antibody specific for REIC/Dkk-3 protein. Pairs of cancerous and adjacent noncancerous tissues were obtained from 20 patients with RCCC. Of them 17 and 7 cases were analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction, and by Western blot analysis and/or immunohistochemical analysis, respectively.Results: The decreased expression of REIC/Dkk-3 mRNA and protein in human RCCC cell lines, and the specificity of the new antibody were confirmed. In a real-time quantitative reverse transcriptase-polymerase chain reaction study using 17 pairs of RCCC and adjacent normal tissues REIC/Dkk-3 mRNA levels were significantly decreased in carcinoma tissues (by 25% to approximately 95% in 15 pairs). Western blot analysis and immunohistochemistry revealed a significant decrease in REIC/Dkk-3 protein levels in 6 of the 7 and 13 of the 14 RCCC cases analyzed, respectively.Conclusions: The decrease in REIC/Dkk-3 mRNA and protein levels was observed irrespective of tumor grade and stage, indicating the involvement of REIC/Dkk-3 in an initial step of malignant conversion. Consequently REIC/Dkk-3 could be a new molecular target for therapeutic measures against RCCC.