Dicer is down-regulated in clear cell renal cell carcinoma and in vitro Dicer knockdown enhances malignant phenotype transformation

被引:8
|
作者
Ma, Xin [1 ]
Fan, Yang [1 ]
Gao, Yu [1 ]
Zhang, Yu [1 ]
Huang, Qingbo [1 ]
Ai, Qing [1 ]
Ni, Dong [2 ]
Chen, Weihao [1 ]
Zhang, Peng [1 ]
Song, Erlin [1 ]
Wang, Baojun [1 ]
Shi, Taoping [1 ]
Zheng, Tao [1 ]
Zhang, Xu [1 ]
机构
[1] Chinese Peoples Liberat Army Gen Hosp, Mil Postgrad Med Coll, Dept Urol, State Key Lab Kidney Dis, Beijing, Peoples R China
[2] Nankai Univ, Sch Med, Tianjin 300071, Peoples R China
基金
中国国家自然科学基金;
关键词
Dicer; Clear cell renal cell carcinoma; Metastasis; Proliferation; Migration; Invasion; MICRORNA EXPRESSION; POSTTRANSCRIPTIONAL REGULATION; CANCER; METASTASIS; MIRNAS;
D O I
10.1016/j.urolonc.2013.06.011
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Objectives: Although emerging evidence has shown that the deregulation of tnicro-ribonucleic acid (RNA) biogenesis machinery is involved in various human malignancies, this role has not been investigated in clear cell renal cell carcinoma (ccRCC). This study aims to determine whether Dicer, a key enzyme responsible for biogenesis of microRNA, is deregulated in ccRCC. The biological roles of Dicer in vitro are also determined. Materials and methods: The expression of Dicer at messenger RNA and protein levels was detected by real-time quantitative polymerase chain reaction and western blot, respectively, in human kidney tubule epithelial cell line, nonmetastatic 786-0 ceRCC cell line, and metastatic ACI IN ccRCC cell line, as well as in 42 cases of ccRCC surgical specimens including 14 cases with distant metastasis and their corresponding adjacent normal renal tissues. Dicer expression levels in specimens were also measured by immunohistochemical staining. Knockdown of Dicer expression in 786-0 and ACHN ccRCC cell lines was achieved by transfecting short interfering RNA against Dicer. The effects of Dicer on cell proliferation, migration, and invasion were detected by 3-(4,5-dimethylthiazol-2-y1)-5-(3-carboxymethoxypheny1)-2-0-sulfophenyD-211-tetrazolium, inner salt (MTS) assay, flow cytometric analyses, and Boyden chamber Transwell assay, respectively. Results: Compared with human kidney tubule epithelial cell line. Dicer expression levels were significantly down-regulated in 786-0 and ACHN ccRCC cell lines, with the metastatic ACHN ccRCC cell line having even lower levels. Meanwhile. Dicer expression levels were significantly down-regulated in ccRCC surgical specimens compared with adjacent normal renal tissues, with the metastatic ones further reduced, and Dicer messenger RNA levels were significantly correlated with overall tumor-node-metastasis stage of ccRCC. In vitro, the knockdown of Dicer significantly promoted cell proliferation, migration, and invasion. Conclusions: Reduced expression of Dicer may play a role in the tumorigenesis of ccRCC and further decline may be associated with distant metastasis of ccRCC. (C) 2014 Elsevier Inc. All rights reserved.
引用
收藏
页码:46.e9 / 46.e17
页数:9
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