microRNA-210-3p depletion by CRISPR/Cas9 promoted tumorigenesis through revival of TWIST1 in renal cell carcinoma.
microRNA-210-3p depletion by CRISPR/Cas9 promoted tumorigenesis through revival of TWIST1 in renal cell carcinoma.
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DOI：
10.18632/oncotarget.14930
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发表时间：
2017-03-28
期刊：
影响因子：
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通讯作者：
Enokida H
中科院分区：
文献类型：
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作者：
Yoshino H;Yonemori M;Miyamoto K;Tatarano S;Kofuji S;Nohata N;Nakagawa M;Enokida H
先前的研究表明，5种miRNA（miR-885- 5 p、miR-1274、miR-210- 3 p、miR-224和miR-1290）在肾透明细胞癌（ccRCC）中上调最多。我们的重点是从临床角度理解上调miR-210- 3 p的功能后果。为此，我们利用CRISPR/Cas9基因编辑系统耗尽RCC细胞系（786-o、A498和Caki 2）中的miR-210- 3 p并表征结果。我们观察到miR-210- 3 p缺失显著增加了肿瘤发生，包括以上皮-间质转化（EMT）特征性的方式改变A498和Caki 2细胞的形态。这些结果得到了体内异种移植研究的证实，该研究显示来自miR-210- 3 p耗尽的A498细胞的肿瘤生长增强。我们确定Twist相关蛋白1（TWIST 1）是miR-210- 3 p的关键靶点。对癌症基因组图谱数据库中ccRCC患者数据的分析显示，miR-210- 3 p和TWIST 1表达之间呈负相关。与低TWIST 1和高miR-210- 3 p表达相比，高TWIST 1和低miR-210 - 3 p表达与较差的总体生存期和无病生存期相关。这些发现表明，肾细胞癌的进展是由miR-210- 3 p介导的TWIST 1抑制促进的。
Previous studies showed that five miRNAs (miR-885-5p, miR-1274, miR-210-3p, miR-224 and miR-1290) were upregulated the most in clear cell renal cell carcinoma (ccRCC). Our focus was to understand from a clinical standpoint the functional consequences of upregulating miR-210-3p. Towards this, we utilized the CRISPR/Cas9 gene editing system to deplete miR-210-3p in RCC cell lines (786-o, A498 and Caki2) and characterized the outcomes. We observed that miR-210-3p depletion dramatically increased tumorigenesis, including altering the morphology of A498 and Caki2 cells in a manner characteristic of epithelial-mesenchymal transition (EMT). These results were corroborated by in vivo xenograft studies, which showed enhanced growth of tumors from miR-210-3p-depleted A498 cells. We identified Twist-related protein 1 (TWIST1) as a key target of miR-210-3p. Analysis of the ccRCC patient data in The Cancer Genome Atlas database showed a negative correlation between miR-210-3p and TWIST1 expression. High TWIST1 and low miR-210-3p expression associated with poorer overall and disease-free survival as compared to low TWIST1 and high miR-210-3p expression. These findings suggest that renal cell carcinoma progression is promoted by TWIST1 suppression mediated by miR-210-3p.