MicroRNA-31 controls G protein alpha-13 (GNA13) expression and cell invasion in breast cancer cells.
MicroRNA-31 controls G protein alpha-13 (GNA13) expression and cell invasion in breast cancer cells.
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DOI：
10.1186/s12943-015-0337-x
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发表时间：
2015-03-26
期刊：
影响因子：
37.3
通讯作者：
Casey PJ
中科院分区：
文献类型：
--
作者：
Rasheed SA;Teo CR;Beillard EJ;Voorhoeve PM;Zhou W;Ghosh S;Casey PJ
Gα13（GNA 13）是异源三聚体G蛋白的α亚基，通过特异性G蛋白偶联受体（GPCR）介导信号传导。我们最近的研究表明，通过特定的microRNAs（miRNAs或miRs）控制GNA 13的表达对于前列腺癌细胞的侵袭是重要的。然而，对乳腺癌中GNA 13表达的控制知之甚少。进行该项目以确定（i）增强的GNA 13表达对于乳腺癌细胞侵袭是否重要，以及（ii）如果是，则确定乳腺癌中GNA 13表达失调的机制。为了确定可能调控GNA 13的miRNA，使用在线miRNA靶标预测工具Targetscan和GNA 13 -3′-UTR的荧光素酶测定。采用RT-PCR、Western blotting和体外Boyden chamber法研究miRNAs对GNA 13 mRNA、蛋白表达及侵袭能力的影响。使用MTT测定进行细胞增殖。GNA 13在MCF-10a细胞中的过表达诱导侵袭，而在MDA-MB-231细胞中GNA 13表达的敲低抑制侵袭。预测与GNA 13的3′-UTR结合的miRNA的表达分析显示，miR-31在乳腺癌细胞中表现出与GNA 13蛋白表达的负相关性。miR-31在MDA-MB-231细胞中的异位表达显著降低了GNA 13 mRNA和蛋白水平，以及GNA 13 -3′-UTR报告基因活性。相反，在MCF-10a细胞中阻断miR-31活性可诱导GNA 13 mRNA、蛋白和3′-UTR报告基因活性。此外，miR-31的表达显著抑制了MDA-MB-231细胞的侵袭，并且这种作用部分地被这些细胞中GNA 13的异位表达所挽救。对48例人乳腺癌组织的检查显示，GNA 13 mRNA水平与miR-31水平呈负相关。这些数据提供了强有力的证据表明，乳腺癌细胞中GNA 13的表达是由涉及miR-31的转录后机制调控的。此外，我们的数据显示，miR-31部分通过靶向乳腺癌细胞中的GNA 13表达来调节乳腺癌细胞侵袭。miR-31表达缺失和GNA 13表达增加可作为乳腺癌进展的生物标志物。本文的在线版本（doi：10.1186/s12943-015-0337-x）包含补充材料，可供授权用户使用。
Gα13 (GNA13) is the α subunit of a heterotrimeric G protein that mediates signaling through specific G protein-coupled receptors (GPCRs). Our recent study showed that control of GNA13 expression by specific microRNAs (miRNAs or miRs) is important for prostate cancer cell invasion. However, little is known about the control of GNA13 expression in breast cancers. This project was carried out to determine (i) whether enhanced GNA13 expression is important for breast cancer cell invasion, and (ii) if so, the mechanism of deregulation of GNA13 expression in breast cancers. To determine the probable miRNAs regulating GNA13, online miRNA target prediction tool Targetscan and Luciferase assays with GNA13-3′-UTR were used. Effect of miRNAs on GNA13 mRNA, protein and invasion was studied using RT-PCR, western blotting and in vitro Boyden chamber assay respectively. Cell proliferation was done using MTT assays. Overexpression of GNA13 in MCF-10a cells induced invasion, whereas knockdown of GNA13 expression in MDA-MB-231 cells inhibited invasion. Expression analysis of miRNAs predicted to bind the 3′-UTR of GNA13 revealed that miR-31 exhibited an inverse correlation to GNA13 protein expression in breast cancer cells. Ectopic expression of miR-31 in MDA-MB-231 cells significantly reduced GNA13 mRNA and protein levels, as well as GNA13-3′-UTR-reporter activity. Conversely, blocking miR-31 activity in MCF-10a cells induced GNA13 mRNA, protein and 3′-UTR reporter activity. Further, expression of miR-31 significantly inhibited MDA-MB-231 cell invasion, and this effect was partly rescued by ectopic expression of GNA13 in these cells. Examination of 48 human breast cancer tissues revealed that GNA13 mRNA levels were inversely correlated to miR-31 levels. These data provide strong evidence that GNA13 expression in breast cancer cells is regulated by post-transcriptional mechanisms involving miR-31. Additionally our data shows that miR-31 regulates breast cancer cell invasion partially via targeting GNA13 expression in breast cancer cells. Loss of miR-31 expression and increased GNA13 expression could be used as biomarkers of breast cancer progression. The online version of this article (doi:10.1186/s12943-015-0337-x) contains supplementary material, which is available to authorized users.
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