Aberrant expression of microRNAs involved in epithelial-mesenchymal transition of HT-29 cell line
Aberrant expression of microRNAs involved in epithelial-mesenchymal transition of HT-29 cell line
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DOI：
10.1002/cbin.10087
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发表时间：
2013-07-01
影响因子：
3.9
通讯作者：
Xu, Xiao-Ping
中科院分区：
文献类型：
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作者：
Cai, Zhi-Gang;Zhang, Shao-Ming;Xu, Xiao-Ping
上皮-间质转化（EMT）是肿瘤转移的重要环节。microRNAs（miRNAs）是一类在转录后调控靶基因表达的非编码小分子RNA。miRNA在HT-29结肠细胞EMT中的表达和功能仍然难以捉摸。本研究观察了miRNA在EMT中的表达，并探讨了miRNA对HT-29细胞系EMT的影响。用TGF处理HT-29以建立EMT模型，其中通过实时PCR（qPCR）分析动态调节一系列miRNA。其中，miR-21和miR-27表达显著上调，而miR-22、miR-26、miR-30、miR-181、miR-200 b、miR-200 c和miR-214表达显著下调。使用miRNA抑制剂敲除HT-29中的miRNA，并通过qPCR测定EMT标记物以监测miRNA对EMT过程的影响。结果显示，miR-22不能改变EMT标志物的表达，而miR-200 b的敲低可显著增加上皮标志物N-cadherin、Vimentin、-Sma和Twist 1的表达，降低间充质标志物E-cadherin的表达。生物信息学分析和Western blot结果显示，miR-200 b可直接抑制ZEB 1的表达。总之，miR-200 b通过抑制HT-29中ZEB 1的表达，在TGF -诱导的HT-29 EMT中受到动态调节。
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 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, -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 -induced EMT of HT-29 and miR-200b was essential for EMT by suppressing the expression of ZEB1 in HT-29.