在非小细胞肺癌中，miR-193a的病理表达降低通过靶向WT1-E-钙黏蛋白轴促进转移。

Pathologically decreased expression of miR-193a contributes to metastasis by targeting WT1-E-cadherin axis in non-small cell lung cancers.

作者信息

Chen Junjie, Gao Shenmeng, Wang Chunjing, Wang Zhonggai, Zhang Huxiang, Huang Kate, Zhou Bin, Li Haiying, Yu Zhijie, Wu Jianbo, Chen Chengshui

机构信息

Department of Respiration, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Ouhai District, Wenzhou, 325000, Zhejiang Province, China.

Laboratory of Internal Medicine, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang, Ouhai District, Wenzhou, Zhejiang Province, China.

出版信息

J Exp Clin Cancer Res. 2016 Nov 7;35(1):173. doi: 10.1186/s13046-016-0450-8.

DOI:10.1186/s13046-016-0450-8

PMID:27821145

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5100283/

Abstract

BACKGROUND

The metastatic cascade is a complex and multistep process with many potential barriers. Recently, miR-193a has been reported to be a suppressive miRNA in multiple types of cancers, but its underlying anti-oncogenic activity in non-small cell lung cancers (NSCLC) is not fully elucidated.

METHODS

The expressions of miR-193a (miR-193a-5p) in human lung cancer tissues and cell lines were detected by real-time PCR. Dual-luciferase reporter assay was used to identify the direct target of miR-193a. Cell proliferation, apoptosis, and metastasis were assessed by CCK-8, flow cytometry, and Transwell assay, respectively.

RESULTS

The expression of miR-193a in lung cancer tissues was decreased comparing to adjacent non-tumor tissues due to DNA hypermethylation in lung cancer tissues. Ectopic expression of miR-193a inhibited cell proliferation, colony formation, migration, and invasion in A549 and H1299 cells. Moreover, overexpression of miR-193a partially reversed tumor growth factor-β1 (TGF-β1)-induced epithelial-to-mesenchymal transition (EMT) in NSCLC cells. Mechanistically, miR-193a reduced the expression of WT1, which negatively regulated the protein level of E-cadherin, suggesting that miR-193a might prevent EMT via modulating WT1-E-cadherin axis. Importantly, knockdown of WT1 resembled the anti-cancer activity by miR-193a and overexpression of WT1 partially reversed miR-193a-induced anti-cancer activity, indicating that WT1 plays an important role in miR-193a-induced anti-cancer activity. Finally, overexpression of miR-193a decreased the growth of tumor xenografts in mice.

CONCLUSION

Collectively, our results have revealed an important role of miR-193a-WT1-E-cadherin axis in metastasis, demonstrated an important molecular cue for EMT, and suggested a therapeutic strategy of restoring miR-193a expression in NSCLC.

摘要

背景

转移级联是一个复杂的多步骤过程，存在许多潜在障碍。最近，据报道miR-193a在多种癌症中是一种抑制性miRNA，但其在非小细胞肺癌（NSCLC）中的潜在抗癌活性尚未完全阐明。

方法

通过实时PCR检测人肺癌组织和细胞系中miR-193a（miR-193a-5p）的表达。使用双荧光素酶报告基因检测法鉴定miR-193a的直接靶点。分别通过CCK-8、流式细胞术和Transwell检测评估细胞增殖、凋亡和转移。

结果

由于肺癌组织中的DNA高甲基化，与相邻非肿瘤组织相比，肺癌组织中miR-193a的表达降低。miR-193a的异位表达抑制了A549和H1299细胞的增殖、集落形成、迁移和侵袭。此外，miR-193a的过表达部分逆转了肿瘤生长因子-β1（TGF-β1）诱导的NSCLC细胞上皮-间质转化（EMT）。机制上，miR-193a降低了WT1的表达，WT1负向调节E-钙黏蛋白的蛋白水平，表明miR-193a可能通过调节WT1-E-钙黏蛋白轴来预防EMT。重要的是，敲低WT1类似于miR-193a的抗癌活性，而WT1的过表达部分逆转了miR-193a诱导的抗癌活性，表明WT1在miR-193a诱导的抗癌活性中起重要作用。最后,miR-193a的过表达降低了小鼠肿瘤异种移植瘤的生长。