上调的 microRNA miR-21 通过抑制 KIBRA 和 Hippo 信号通路促进肺腺癌的进展。

Upregulated microRNA miR-21 promotes the progression of lung adenocarcinoma through inhibition of KIBRA and the Hippo signaling pathway.

机构信息

Department of Respiratory Diseases, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Zhengzhou, 450000, Henan, China.

出版信息

Biomed Pharmacother. 2018 Dec;108:1845-1855. doi: 10.1016/j.biopha.2018.09.125. Epub 2018 Oct 20.

DOI:10.1016/j.biopha.2018.09.125

PMID:30372890

Abstract

PURPOSE

In this study, we aimed to identify the key pathways and hub genes in lung adenocarcinoma (LAD) through bioinformatics analysis and to identify the miRNAs that targeted the selected hub gene. The present study was conducted to explore the effect of the hub gene KIBRA, the Hippo signaling pathway and miR-21 on LAD progression.

METHODS

Through gene set enrichment analysis (GSEA), the enriched KEGG pathways involved in LAD were identified. Weighted correlation network analysis (WGCNA) was employed to screen out hub genes. The differentially expressed miRNAs related to the hub gene were then screened by the network analysis. The mRNA expression levels of miR-21 and KIBRA were detected by qRT-PCR. The protein expression levels of KIBRA and the pathway related proteins LATS2 and YAP were determined by Western blot assay. The target relationship between miR-21 and KIBRA was confirmed by the dual luciferase reporter assay. Through colony formation assay, the viability of the LAD cells was determined. In addition, the mobility of LAD cells was detected by wound healing assays, and flow cytometry was employed to detect apoptotic cancer cells.

RESULTS

The hub gene identified in the black module was KIBRA, and suppression of the Hippo signaling pathway was detected in LAD. KIBRA was downregulated and miR-21 was upregulated in LAD tissues and cells; moreover, miR-21 was found to target KIBRA. KIBRA reduced the proliferative and invasive ability of LAD cells and induced apoptosis. KIBRA also activated the Hippo signaling pathway in LAD. The role of MiR-21 was opposite that of KIBRA in LAD.

CONCLUSION

MiR-21 suppressed the Hippo signaling pathway and promoted the progression of LAD through targeting KIBRA.

摘要

目的

本研究通过生物信息学分析，旨在鉴定肺腺癌（LAD）中的关键途径和枢纽基因，并鉴定靶向选定枢纽基因的 miRNAs。本研究旨在探讨枢纽基因 KIBRA、Hippo 信号通路和 miR-21 对 LAD 进展的影响。

方法

通过基因集富集分析（GSEA），鉴定与 LAD 相关的富集 KEGG 途径。采用加权相关网络分析（WGCNA）筛选枢纽基因。然后通过网络分析筛选与枢纽基因相关的差异表达 miRNA。通过 qRT-PCR 检测 miR-21 和 KIBRA 的 mRNA 表达水平。通过 Western blot 检测 KIBRA、通路相关蛋白 LATS2 和 YAP 的蛋白表达水平。通过双荧光素酶报告基因实验验证 miR-21 和 KIBRA 之间的靶关系。通过集落形成实验测定 LAD 细胞的活力。此外，通过划痕愈合实验检测 LAD 细胞的迁移能力，并用流式细胞术检测凋亡癌细胞。

结果

黑色模块中鉴定的枢纽基因是 KIBRA，并且在 LAD 中检测到 Hippo 信号通路被抑制。KIBRA 在 LAD 组织和细胞中下调，miR-21 上调；此外，发现 miR-21 靶向 KIBRA。KIBRA 降低了 LAD 细胞的增殖和侵袭能力，并诱导细胞凋亡。KIBRA 还激活了 LAD 中的 Hippo 信号通路。miR-21 在 LAD 中的作用与 KIBRA 相反。