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微小 RNA-17 在后转录水平上调节多囊肾病 2 基因并促进细胞增殖。

MicroRNA-17 post-transcriptionally regulates polycystic kidney disease-2 gene and promotes cell proliferation.

机构信息

Core Facility of Genetically Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, 1# Keyuan the Fourth Road, The District of Hi&Tech, Chengdu, Sichuan 610041, People's Republic of China.

出版信息

Mol Biol Rep. 2010 Jul;37(6):2951-8. doi: 10.1007/s11033-009-9861-3. Epub 2009 Oct 10.

DOI:10.1007/s11033-009-9861-3
PMID:19821056
Abstract

To identify the possible microRNAs (miRNAs) which target the polycystic kidney disease-2 gene (PKD2), and clarify effects of the miRNAs on PKD2. We preliminarily used bioinformatics to analyze 3'UTR (3'untranslated regions) of PKD1 and PKD2 in order to predict the potential microRNAs targeted on them. Subsequently, the stable cell lines with overexpression of microRNA-17 (miR-17) were screened, and luciferase assay combined with the mutation 3'UTR of PKD2 were performed to verify PKD2 is the target of miR-17. Moreover, RT-PCR and Western Blotting were used to determine the post-transcriptionally regulation of PKD2 by miR-17. Finally, MTT cell assays allied with PKD2 rescued strategy were employed to evaluate cell proliferation effects. Our study firstly found that the 3'UTR of PKD2 was more conservation than that of PKD1, and microRNA-17 directly targets the 3'UTR of PKD2 and post-transcriptionally repress the expression of PKD2. Moreover, our findings also demonstrated that overexpression of miR-17 may promote cell proliferation via post-transcriptionally repression of PKD2 in HEK 293T. This suggested that microRNA might be a novel mechanism for cystogenesis as well as a potential therapeutic target for the cell proliferation of autosomal dominant polycystic kidney disease (ADPKD).

摘要

为了鉴定可能靶向多囊肾病 2 基因(PKD2)的 microRNAs(miRNAs),并阐明这些 miRNAs 对 PKD2 的影响。我们初步使用生物信息学方法分析了 PKD1 和 PKD2 的 3'UTR(非翻译区),以预测它们可能靶向的潜在 microRNAs。随后,筛选出 miRNA-17(miR-17)过表达的稳定细胞系,并通过荧光素酶测定和 PKD2 3'UTR 突变验证 PKD2 是 miR-17 的靶基因。此外,还通过 RT-PCR 和 Western Blotting 确定了 miR-17 对 PKD2 的转录后调控作用。最后,采用 MTT 细胞测定和 PKD2 挽救策略来评估细胞增殖效应。本研究首次发现 PKD2 的 3'UTR 比 PKD1 的更保守,miR-17 直接靶向 PKD2 的 3'UTR,并通过转录后抑制 PKD2 的表达。此外,我们的研究结果还表明,miR-17 的过表达可能通过 PKD2 的转录后抑制促进 HEK 293T 细胞的增殖。这表明 microRNA 可能是多囊肾病发生的一种新机制,也是常染色体显性多囊肾病(ADPKD)细胞增殖的潜在治疗靶点。

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