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微小RNA-99b-5p靶向卷曲蛋白8以抑制非小细胞肺癌的增殖、迁移和侵袭。

miRNA-99b-5p targets FZD8 to inhibit non-small cell lung cancer proliferation, migration and invasion.

作者信息

Liu Rui, Chen Yajuan, Shou Tao, Hu Jing, Qing Chen

机构信息

Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming 650031, People's Republic of China,

Department of Oncology, The First People's Hospital of Yunnan Province, Kunming 650032, Yunnan, People's Republic of China.

出版信息

Onco Targets Ther. 2019 Apr 8;12:2615-2621. doi: 10.2147/OTT.S199196. eCollection 2019.

DOI:10.2147/OTT.S199196
PMID:31040702
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6459141/
Abstract

BACKGROUND

miRNAs were found to play crucial roles in regulating cellular behaviors. The aim of this study was to investigate the biological function of miRNA-99b-5p (miR-99b-5p) in non-small cell lung cancer (NSCLC).

MATERIALS AND METHODS

miR-99b-5p expression level in NSCLC cell lines was detected by quantitative real-time PCR (qRT-PCR). Cell proliferation, migration and invasion were examined by cell counting kit-8 (CCK-8) assay, wound-healing assay and Transwell invasion assay, respectively. Dual-luciferase activity reporter assay and Western blot assay were conducted to validate the target of miR-99b-5p.

RESULTS

The expression of miR-99b-5p was decreased in NSCLC cell lines compared with normal cell line. Overexpression of miR-99b-5p inhibits cell proliferation, migration and invasion in vitro. FZD8 was validated as a direct target of miR-99b-5p. Overexpression of FZD8 partially abolished the effects of miR-99b-5p mimic on NSCLC cell behaviors.

CONCLUSION

Collectively, our results demonstrated that miR-99b-5p inhibits NSCLC cell proliferation, migration and invasion through targeting FZD8. This newly identified miR-99b-5p/FZD8 axis provided novel insights into the mechanisms underlying NSCLC progression.

摘要

背景

微小RNA（miRNA）被发现可在调节细胞行为中发挥关键作用。本研究旨在探讨miRNA - 99b - 5p（miR - 99b - 5p）在非小细胞肺癌（NSCLC）中的生物学功能。

材料与方法

采用定量实时聚合酶链反应（qRT - PCR）检测NSCLC细胞系中miR - 99b - 5p的表达水平。分别通过细胞计数试剂盒 - 8（CCK - 8）检测、伤口愈合检测和Transwell侵袭检测来检测细胞增殖、迁移和侵袭情况。进行双荧光素酶活性报告基因检测和蛋白质免疫印迹法（Western blot）检测以验证miR - 99b - 5p的靶标。

结果

与正常细胞系相比，NSCLC细胞系中miR - 99b - 5p的表达降低。miR - 99b - 5p的过表达在体外抑制细胞增殖、迁移和侵袭。已证实卷曲蛋白8（FZD8）是miR - 99b - 5p的直接靶标。FZD8的过表达部分消除了miR - 99b - 5p模拟物对NSCLC细胞行为的影响。

结论

总体而言，我们的结果表明miR - 99b - 5p通过靶向FZD8抑制NSCLC细胞增殖、迁移和侵袭。这个新发现的miR - 99b - 5p/FZD8轴为NSCLC进展的潜在机制提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/dc555608ce3a/ott-12-2615Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/39fc5b45da89/ott-12-2615Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/ead7cb8b0cf6/ott-12-2615Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/43b4fa6b4a91/ott-12-2615Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/dc555608ce3a/ott-12-2615Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/39fc5b45da89/ott-12-2615Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/ead7cb8b0cf6/ott-12-2615Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/43b4fa6b4a91/ott-12-2615Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdb1/6459141/dc555608ce3a/ott-12-2615Fig4.jpg

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