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Aberrant expression of microRNAs involved in epithelial-mesenchymal transition of HT-29 cell line.

机译:参与HT-29细胞系上皮-间质转化的microRNA异常表达。

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Epithelial-mesenchymal transition (EMT) is an essential step for cancer metastasis. MicroRNAs (miRNAs) are small non-coding RNAs that regulate target-mRNAs post-transcriptionally. The expression and function of miRNAs in EMT of HT-29 colonic cells remain elusive. This study looks at expression of miRNAs in EMT and explores the effects of miRNAs on EMT in HT-29 cell line. HT-29 was treated with TGF β to establish an EMT model, in which a collection of miRNAs was dynamically regulated by real-time PCR (qPCR) analysis. Among them, miR-21 and miR-27 were significantly upregulated, while miR-22, miR-26, miR-30, miR-181, miR-200b, miR-200c and miR-214 were markedly downregulated. MiRNA-inhibitors were used to knockdown miRNAs in HT-29 and EMT markers were determined by qPCR to monitor the effects of miRNAs on EMT process. Results showed that miR-22 could not alter the expression of EMT markers, while knockdown of miR-200b could significantly increase that of epithelial markers, N-cadherin, Vimentin, α-Sma and Twist1 and decrease that of mesenchymal marker, E-cadherin. Bioinformatic analysis and Western blot showed that ZEB1 was directly suppressed by miR-200b. In conclusion, miRNAs are dynamically regulated in TGF β-induced EMT of HT-29 and miR-200b was essential for EMT by suppressing the expression of ZEB1 in HT-29.
机译:上皮-间质转化(EMT)是癌症转移的重要步骤。 MicroRNA(miRNA)是小的非编码RNA,可在转录后调节靶标mRNA。 miRNA在HT-29结肠细胞EMT中的表达和功能仍然难以捉摸。这项研究着眼于EMT中miRNA的表达,并探讨了HT-29细胞系中miRNA对EMT的影响。用TGFβ处理HT-29以建立EMT模型,其中通过实时PCR(qPCR)分析动态调节miRNA的集合。其中,miR-21和miR-27显着上调,而miR-22,miR-26,miR-30,miR-181,miR-200b,miR-200c和miR-214显着下调。使用miRNA抑制剂敲低HT-29中的miRNA,并通过qPCR确定EMT标记,以监测miRNA对EMT过程的影响。结果表明,miR-22不能改变EMT标记的表达,而敲低miR-200b则可以显着增加上皮标记N-钙黏着蛋白,波形蛋白,α-Sma和Twist1的表达,而降低间充质标记E-钙黏着蛋白的表达。 。生物信息学分析和Western印迹表明,miR-200b直接抑制ZEB1。总之,miRNA在TGFβ诱导的HT-29的EMT中被动态调节,而miR-200b通过抑制HT-29中ZEB1的表达对EMT至关重要。

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