Mir-186-5p通过靶向TCF4转录因子调控WNT信号通路

Z. Bayat, B. Soltani
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引用次数: 0

摘要

进化上保守的Wnt信号通路在胚胎发育、组织稳态和分化过程中发挥着重要作用。这种途径在许多癌症中被解除调节,尤其是在癌症中。miRNA是一类小型非编码RNA,可能在许多基因和信号通路(如WNT信号通路)的转录后调控中发挥重要作用。在这里,我们打算研究miR-186-5p是否能够通过抑制TCF4基因表达来调节WNT信号通路。miR-186-5p被生物信息学预测为TCF4基因表达的候选调节因子,然后,在本实验研究中,miR-186-5p在SW480细胞系中过表达,并通过定量逆转录聚合酶链反应(RT-qPCR)检测其表达增加。在SW480细胞系中用TOP/FOP闪烁分析法分析了miR-186-5p对WNT通路的影响。最后,用流式细胞仪研究了miR-186-5p过表达对SW480细胞系细胞周期进程的影响。miR-186-5p在SW480细胞系中过表达,其过表达导致TCF4 mRNA水平显著降低。TOP/FOP快速测定证实了miR-186-5p对SW480细胞中Wnt途径的负面影响。最后,通过流式细胞术检测,miR186-5p在SW480细胞中的过表达导致细胞周期停滞在亚G1期。总之,累积结果表明,靶向TCF4的miR-186-5p可能是WNT信号通路的调节因子之一。
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Mir-186-5p Regulates WNT Signaling Pathway by Targeting TCF4 Transcription Factor
The evolutionarily conserved Wnt signaling pathway plays essential roles during embryonic development, tissue homeostasis and differentiation. This pathway is deregulated in many cancers especially colorectal cancer. MiRNAs are a class of small noncoding RNAs that may play a major role in post transcriptional regulation of many genes and signaling pathway such as WNT signaling pathway. Here, we intended to investigate if miR-186-5p is capable of regulating WNT signaling pathway wia suppression TCF4 gene expression. miR-186-5p was bioinformatically predicted as a candidate regulator of TCF4 gene expression and then, in this experimental study, miR-186-5p was overexpressed in SW480 cell line and its increased expression was detected through quantitative reverse transcription polymerase chain reaction (RT-qPCR). The effect of miR-186-5p on WNT pathway was analysied with TOP/FOP flash assay in SW480 cell line. Finally, flow cytometery was used to inves tigate the effect of miR-186-5p overexpression on cell cycle progression in SW480 cell line. miR-186-5p was overexpressed in the SW480 cell line and its overexpression resulted in significant reduction of the TCF4 mRNA level. TOP/FOP flash assay, confirmed the negative effect of miR-186-5p on the Wnt pathway in SW480 cells. Finally, Overexpression of miR186-5p in SW480 cells resulted in cell cycle arrest in subG1 phase, detected by flow cytometry. Overall, accumulative results indi-cated that miR-186-5p by targeting TCF4 is potentially one of the regulators of the WNT signaling pathway.
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