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原代肺癌细胞条件培养液通过 TGF-β1 和 miRNA21 合作诱导 A549 肺癌细胞系 EMT。

Conditioned medium of primary lung cancer cells induces EMT in A549 lung cancer cell line by TGF-ß1 and miRNA21 cooperation.

机构信息

SC Cell Biology and Biotherapy, Istituto Nazionale Tumori IRCCS, Fondazione G. Pascale, Naples, Italy.

reiThera srl, Rome, Italy.

出版信息

PLoS One. 2019 Jul 25;14(7):e0219597. doi: 10.1371/journal.pone.0219597. eCollection 2019.

DOI:10.1371/journal.pone.0219597
PMID:31344049
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6657837/
Abstract

The epithelial-mesenchymal transition (EMT) plays a key role in tumor progression, drug resistance and metastasis. Recently, numerous microRNA (miRNA) have been described to regulate EMT in tumor progression. In this study, we found that conditioned medium from the LC212 non-small-cell lung cancer (NSCLC) cell line (LC212-CM) induces morphological changes and overexpression of Vimentin, CD90, SMAD 2/3, SLUG and TWIST in A549 NSCLC cells, consistent with a mesenchymal phenotype. To identify the soluble mediators in LC212-CM involved in this phenomenon, we performed miRNA profiling and TGF-β1 quantification. We found that LC212-CM contains high levels of TGF-β1 as well as different secreted miRNAs. We focused our attention on Homo sapiens-microRNA21 (hsa-miR21), one of most relevant miRNA associated with lung cancer progression, metastasis and EMT. An hsa-miR21 antagomiR was able to prevent the LC212-CM-induced EMT phenotype in A549 cells. Furthermore, we found that TGF-β1 and hsa-miR21 cooperate in the induction of EMT in A549 cells. Intriguingly, TGF-β1 was found to induce hsa-miR21 expression in A549 cell, thus suggesting that the hsa-miR21 mediates at least in part the pro-EMT effects of TGF-β1. In conclusion, hsa-miR21 and TGF-β1 are involved in autocrine and paracrine circuits that regulate the EMT status of lung cancer cells.

摘要

上皮-间充质转化（EMT）在肿瘤进展、耐药性和转移中起着关键作用。最近，大量的 microRNA（miRNA）被描述为调节肿瘤进展中的 EMT。在本研究中，我们发现 LC212 非小细胞肺癌（NSCLC）细胞系的条件培养基（LC212-CM）诱导 A549 NSCLC 细胞发生形态变化和波形蛋白、CD90、SMAD2/3、SLUG 和 TWIST 的过表达，表现出间充质表型。为了鉴定 LC212-CM 中参与这种现象的可溶性介质，我们进行了 miRNA 谱分析和 TGF-β1 定量。我们发现 LC212-CM 含有高水平的 TGF-β1 和不同的分泌 miRNA。我们将注意力集中在 Homo sapiens-microRNA21（hsa-miR21）上，它是与肺癌进展、转移和 EMT 最相关的 miRNA 之一。hsa-miR21 反义寡核苷酸能够阻止 LC212-CM 在 A549 细胞中诱导的 EMT 表型。此外，我们发现 TGF-β1 和 hsa-miR21 合作诱导 A549 细胞 EMT。有趣的是，我们发现 TGF-β1 诱导 A549 细胞中 hsa-miR21 的表达，因此表明 hsa-miR21 至少部分介导了 TGF-β1 的 EMT 促进作用。总之，hsa-miR21 和 TGF-β1 参与调节肺癌细胞 EMT 状态的自分泌和旁分泌循环。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/a3b681823996/pone.0219597.g006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/a3b681823996/pone.0219597.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/84f1a45b8665/pone.0219597.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/358384b85567/pone.0219597.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/96c184725f3f/pone.0219597.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccb/6657837/d7de7c83ba73/pone.0219597.g004.jpg
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