Department of Laboratory Medicine and Keenan Research Centre, Li Ka Shing Knowledge Institute, Toronto, Ontario, Canada.
J Urol. 2010 Feb;183(2):743-51. doi: 10.1016/j.juro.2009.09.086.
miRNAs are small, nonprotein coding RNAs that are differentially expressed in many malignancies. We previously identified 80 miRNAs that are dysregulated in clear cell renal cell carcinoma. In this study we validated over expression of the miR-17-92 cluster in clear cell renal cell carcinoma and tested the effect of 2 members of this cluster (miR-17-5p and miR-20a) on tumor proliferation. We also elucidated the role of miRNA in clear cell renal cell carcinoma pathogenesis with bioinformatics.
miRNA expression was validated by quantitative reverse transcriptase-polymerase chain reaction. The cell proliferation effect of miR-17-5p and miR-20a was tested in a renal adenocarcinoma cell line model. Multiple in silico analyses were done of dysregulated miRNAs.
We validated miR-71-92 cluster over expression in clear cell renal cell carcinoma by quantitative reverse transcriptase-polymerase chain reaction. Transfection of miR-20a inhibitor significantly decreased cell proliferation in a dose dependent manner. Transfection of miR-17-5p, which is not endogenously expressed in the ACHN cell line, led to increased cell proliferation compared to control values. This effect was suppressed by miR-17-5p inhibitor. Bioinformatics analysis identified 10 clusters of miRNAs dysregulated in clear cell renal cell carcinoma that followed the same expression patterns. We also identified matching patterns between reported chromosomal aberration in clear cell renal cell carcinoma and miRNA dysregulation for 37.5% of the miRNAs. Target prediction analysis was done using multiple algorithms. Many key molecules in clear cell renal cell carcinoma pathogenesis, including HIFs, mTOR, VEGF and VHL, were potential targets for dysregulated miRNAs.
A significant number of dysregulated proteins in clear cell renal cell carcinoma are potential miRNA targets. Also, many clear cell renal cell carcinoma dysregulated miRNAs are phylogenetically conserved.
miRNA 是一种小的非编码 RNA，在许多恶性肿瘤中表达差异。我们之前鉴定了 80 个在透明细胞肾细胞癌中失调的 miRNA。在这项研究中，我们验证了 miR-17-92 簇在透明细胞肾细胞癌中的过表达，并测试了该簇的 2 个成员（miR-17-5p 和 miR-20a）对肿瘤增殖的影响。我们还通过生物信息学阐明了 miRNA 在透明细胞肾细胞癌发病机制中的作用。
通过定量逆转录聚合酶链反应验证 miRNA 表达。在肾腺癌细胞系模型中测试 miR-17-5p 和 miR-20a 的细胞增殖效应。对失调 miRNA 进行了多次计算机分析。
我们通过定量逆转录聚合酶链反应验证了 miR-71-92 簇在透明细胞肾细胞癌中的过表达。miR-20a 抑制剂的转染以剂量依赖的方式显著降低细胞增殖。转染 ACHN 细胞系中内源性表达的 miR-17-5p 导致细胞增殖增加，与对照值相比。这种效应被 miR-17-5p 抑制剂抑制。生物信息学分析鉴定出 10 个簇的 miRNA 在透明细胞肾细胞癌中失调，遵循相同的表达模式。我们还发现了透明细胞肾细胞癌中报道的染色体畸变与 miRNA 失调之间的匹配模式，对于 37.5%的 miRNA 是如此。使用多种算法进行了靶预测分析。透明细胞肾细胞癌发病机制中的许多关键分子，包括 HIFs、mTOR、VEGF 和 VHL，都是失调 miRNA 的潜在靶标。
透明细胞肾细胞癌中大量失调的蛋白质是潜在的 miRNA 靶标。此外，许多透明细胞肾细胞癌失调的 miRNA 是系统发育保守的。
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