miRNA-138与EZH2在肾透明细胞癌发生发展中作用机制的研究

秦洁; 李晓庆; 赵春娟; 漆正宇; 郭新; 龙霞; 余振东; 桂耀庭

doi:10.13201/j.issn.1001-1420.2014.09.024

miRNA-138与EZH2在肾透明细胞癌发生发展中作用机制的研究

- 1. 广东省男性生殖与遗传重点实验室, 北京大学深圳医院(广东深圳, 518036);
- 2. 北京大学深圳医院检验科;
- 3. 汕头大学医学院
基金项目:
国家自然科学基金（编号81100471，81270748）；深圳市科技计划项目（编号201303041，201302042，201302043）；深圳市科技研发资金基础研究计划资助项目（编号JCYJ20130402114702127）

详细信息

通讯作者: 秦洁,E-mail:yzqinjie@126.com

中图分类号: R737.11

收稿日期: 2014-03-04

Study on mechanism of miRNA-138 and EZH2 in the occurrence and development of renal clear cell carcinoma

- 1. Key Laboratory of Male Reproductive and Genetic of Guangdong Province, Peking University Shenzhen Hospital, Shenzhen, Guangdong, 518036, China;
- 2. Department of Clinlical Laboratory of Peking University Shenzhen Hospital;
- 3. Shantou University Medical College

More Information

Corresponding author: QIN Jie, E-mail: yzqinjie@126.com

Received Date: 04 March 2014

摘要

摘要: 目的：探讨miRNA-138及其调控靶基因EZH2在肾透明细胞癌（RCC）发生发展中的相关作用机制。方法：利用第二代大规模平行测序技术对10例RCC及对应癌旁组织进行全基因组测序，在肾癌组织中筛选出表达显著上调的miRNA-138；其次在人肾癌细胞株786-O、ACHN、Caki-1、769-P、os-rc-2及对照细胞人胚肾细胞293T中对miRNA-138进行转染，采用划痕实验检测miRNA-138对细胞株侵袭和迁移能力的影响；进而用多种预测软件预测靶基因，对预测的候选靶基因EZH2进行PCR验证。结果：发现miRNA-138在RCC中表达上调，并能够显著促进人肾癌细胞株786-O、ACHN的迁移；而其候选靶基因EZH2的表达在临床组织标本中显著上调；相应蛋白变化需要进一步验证。结论：EZH2表达在RCC中明显高于正常组织，提示EZH2可能是RCC一个敏感的生物学标志。推测miRNA-138与其靶基因EZH2可能参与肿瘤代谢和细胞凋亡等信号通路，在RCC的发生发展中发挥重要作用。
- 肾透明细胞癌 /
- miRNA-138 /
- EZH2
Abstract: Objective: To investigate the mechanism of miRNA-138 and its target gene enhancer of zeste homolog 2 (EZH2) in the occurrence and development of renal clear cell carcinoma (RCC). Method: The second-generation massively parallel signature sequencing examining the miRNA expression in tumor and the corresponding paracancerous tissues of ten RCC patients was used to screen that miRNA-138 was notably up-regulated in RCC tissues. The scratch assay was performed to detect the invasion and migration, after that the renal cell carcinoma cell lines, 786-O, ACHN, Caki-1, 769-P, os-rc-2 and the control cell line 293T were transfected with miRNA-138. Finally bioinformatics analysis was used to predict EZH2 as a candidate target gene and PCR was performed to validate it.Result: MiRNA-138 was significantly up-regulated in RCC and could promote the migration of 786-O and ACHN cell lines remarkably. While its candidate target gene EZH2 was dramatically up-regulated in RCC specimens and its corresponding protein change need further validation. Conclusion: The expression of EZH2 is obviously higher in RCC than in the adjacent tissue, which indicates that EZH2 may be a sensitive biomarker in RCC. MiRNA-138 and its target gene EZH2 may be involved in the tumor metabolism and apoptosis signaling pathways and play important roles in the occurrence and development of RCC.
- renal clear cell carcinoma /
- miRNA-138 /
- enhancer of zeste homolog 2

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参考文献(19)

[1]	Ramana J. RCDB:Renal Cancer Gene Database[J]. BMC Res Notes, 2012, 18(5):246.
[2]	Alexiev B A, Tavora F. Histology and immunohistochemistry of clear cell adenocarcinoma of the urethra:histogenesis and diagnostic problems[J]. Virchows Arch, 2013, 462(2):193-201.
[3]	Wotschofsky Z, Liep J, Meyer H A, et al. Identification of metastamirs as metastasis-associated microRNAs in clear cell renal cell carcinomas[J]. Int J Biol Sci, 2012, 8(10):1363-1374.
[4]	Osanto S, Qin Y, Buermans H P, et al. Genome-Wide MicroRNA expression analysis of clear cell renal cell carcinoma by next generation deep sequencing[J]. PLoS ONE, 2012, 7(6):e38298.
[5]	Girgis A H, Iakovlev V V, Beheshti B, et al. Multilevel whole-genome analysis reveals candidate biomarkers in clear cell renal cell carcinoma[J]. Cancer Res, 2012, 72(20):5273-5284.
[6]	Zhai Q, Zhou L, Zhao C, et al. Identification of miR-508-3p and miR-509-3p that are associated with cell invasion and migration and involved in the apoptosis of renal cell carcinoma[J]. Biochem Biophys Res Commun, 2012, 419(4):621-626.
[7]	Darwish O M, Kapur P,Youssef R F, et al. Cumulative number of altered biomarkers in mammalian target of rapamycin pathway is an Independent predictor of outcome in patients with clear cell renal cell carcinoma[J]. Urology, 2013, 81(3):581-586.
[8]	Wotschofsky Z, Busch J, Jung M, et al. Diagnostic and prognostic potential of differentially expressed miRNAs between metastatic and non-metastatic renal cell carcinoma at the time of nephrectomy[J]. Clin Chim Acta, 2013, 416:5-10.
[9]	Hua Y, Larsen N, Kalyana-Sundaram S, et al. miRConnect 2.0:identification of oncogenic, antagonistic miRNA families in three human cancers[J]. BMC Genomics, 2013, 14(1):179.
[10]	Lin J, Horikawa Y, Tamboli P, et al. Genetic variations in microRNA-related genes are associated with survival and recurrence in patients with renal cell carcinoma[J]. Carcinogenesis, 2010, 31(10):1805-1812.
[11]	Liu H, Brannon A R, Reddy A R, et al. Identifying mRNA targets of microRNA dysregulated in cancer:with application to clear cell renal cell carcinoma[J]. BMC Syst Biol, 2010, 4:51.
[12]	Jung M, Mollenkopf H J, Grimm C, et al. MicroRNA profiling of clear cell renal cell cancer identifies a robust signature to define renal malignancy[J]. J Cell Mol Med, 2009, 13(9B):3918-3928.
[13]	Weng L, Wu X, Gao H, et al. MicroRNA profiling of clear cell renal cell carcinoma by whole-genome small RNA deep sequencing of paired frozen and formalin-fixed, paraffin-embedded t issue specimens[J]. J Pathol, 2010, 222(1):41-51.
[14]	Lee H W, Choe M. Expression of EZH2 in renal cell carcinoma as a novel prognostic marker[J]. Pathol Int, 2012, 62(11):735-741.
[15]	Truax A D, Thakkar M, Greer S F. Dysregulated recruitment of the histone methyltransferase EZH2 to the class II transactivator (CIITA) promoter IV in breast cancer cells[J]. PLoS One, 2012, 7(4):e36013.
[16]	Kong D, Heath E, Chen W, et al. Loss of let-7 up-regulates EZH2 in prostate cancer consistent with the acquisition of cancer stem cell signatures that are attenuated by BR-DIM[J]. PLoS One, 2012, 7(3):e33729.
[17]	成磊. miRNA介导靶基因沉默的分析与识别[D]. 南京:南京航空航天大学,2011.
[18]	Hinz S, Weikert S, Magheli A, et al. Expression profile of the polycomb group protein enhancer of Zeste homologue 2 and its prognostic relevance in renal cell carcinoma[J]. J Urol, 2009, 182(6):2920-2925.
[19]	Qiu S, Huang D, Yin D, et al. Suppression of tumorigenicity by microRNA-138 through inhibition of EZH2-CDK4/6-pRb-E2F1 signal loop in glioblastoma multiforme[J]. Biochim Biophys Acta, 2013, 1832(10):1697-1707.