MiR-130a-3p regulates cell migration and invasion via inhibition of Smad4 in gemcitabine resistant hepatoma cells.
MiR-130a-3p regulates cell migration and invasion via inhibition of Smad4 in gemcitabine resistant hepatoma cells.
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MiR-130a-3p 通过抑制吉西他滨耐药肝癌细胞中的 Smad4 调节细胞迁移和侵袭
DOI：
10.1186/s13046-016-0296-0
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发表时间：
2016-01-27
期刊：
影响因子：
--
通讯作者：
Wu Q
中科院分区：
文献类型：
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作者：
Liu Y;Li Y;Wang R;Qin S;Liu J;Su F;Yang Y;Zhao F;Wang Z;Wu Q
研究背景近年来研究表明，microRNAs（miRNAs）通过抑制其靶基因的表达，在调节细胞生长、侵袭和转移等方面发挥重要作用。据报道，miR-130 a-3 p在多种癌细胞中控制细胞生长、迁移和侵袭。然而，目前尚不清楚miR-130 a-3 p是否调节耐药癌细胞中的上皮-间充质转化（EMT）。因此，本研究旨在探讨miR-130 a-3 p在吉西他滨耐药（GR）肝癌细胞中的作用及其分子机制。进行伤口愈合测定以确定用miR-130 a-3 p模拟物处理的GR HCC细胞中的细胞迁移活性。通过迁移和侵袭实验探讨miR-130 a-3 p在GR HCC细胞中的作用。Western blotting分析检测Smad 4缺失后GR HCC细胞中Smad 4、E-cadherin、Vimentin和MMP-2的表达。进行荧光素酶测定以验证Smad 4是否是miR-130 a-3 p的靶标。结果发现miR-130 a-3 p在GR肝癌细胞中表达下调。此外，我们验证了Smad 4是miR-130 a-3 p的潜在靶点。此外，过表达miR-130 a-3 p抑制Smad 4表达，而抑制miR-130 a-3 p增加Smad 4表达。结论miR-130 a-3 p的过表达或Smad 4的下调抑制了GR HCC细胞的脱落、粘附、迁移和侵袭。因此，激活miR-130 a-3 p或失活Smad 4可能是治疗HCC的新方法。
BackgroundEmerging evidence demonstrates that microRNAs (miRNAs) play an important role in regulation of cell growth, invasion and metastasis through inhibiting the expression of their targets. It has been reported that miR-130a-3p controls cell growth, migration and invasion in a variety of cancer cells. However, it is unclear whether miR-130a-3p regulates epithelial-mesenchymal transition (EMT) in drug resistant cancer cells. Therefore, in the current study, we explore the role and molecular mechanisms of miR-130a-3p in gemcitabine resistant (GR) hepatocellular carcinoma (HCC) cells.MethodsThe real-time RT-PCR was used to measure the miR-130a-3p expression in GR HCC cells compared with their parental cells. The wound healing assay was conducted to determine the cell migratory activity in GR HCC cells treated with miR-130a-3p mimics. The migration and invasion assays were also performed to explore the role of miR-130a-3p in GR HCC cells. Western blotting analysis was used to measure the expression of Smad4, E-cadherin, Vimentin, and MMP-2 in GR HCC cells after depletion of Smad4. The luciferase assay was conducted to validate whether Smad4 is a target of miR-130a-3p. The studentt-test was used to analyze our data.ResultsWe found the down-regulation of miR-130a-3p in GR HCC cells. Moreover, we validate the Smad4 as a potential target of miR-130a-3p. Furthermore, overexpression of miR-130a-3p suppressed Smad4 expression, whereas inhibition of miR-130a-3p increased Smad4 expression. Consistently, overexpression of miR-130a-3p or down-regulation of Smad4 suppressed the cell detachment, attachment, migration, and invasion in GR HCC cells.ConclusionsOur findings provide a molecular insight on understanding drug resistance in HCC cells. Therefore, activation of miR-130a-3p or inactivation of Smad4 could be a novel approach for the treatment of HCC.