Department of Thoracic Surgery, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning, 110032, China.
Cancer Med. 2017 Jun;6(6):1323-1330. doi: 10.1002/cam4.1026. Epub 2017 Apr 24.
The aim of this study was to study the role of miR-372-3p in lung squamous cell carcinoma (LSCC) cell proliferation and invasion by suppressing FGF9. RT-PCR was used to determine miR-372-3p and FGF9 mRNA expression in tissues and cells. Western blot was used to determine FGF9 expression in tissues and NCI-H520 cell line. Dual luciferase reporter gene assay was conducted to confirm that FGF9 can be directly targeted by miR-372-3p. MTT, colony formation assays were conducted to investigate the effects of ectopic miR-372-3p and FGF9 expression on NCI-H520 cell growth. Flow cytometry was used to analyze the influence of miR-372-3p and FGF9 expression on cell cycle distribution and apoptosis. Transwell assay was also conducted to see the effects of miR-372-3p and FGF9 expression on NCI-H520 cell invasiveness. MiR-372-3p was found significantly overexpressed in both LSCC tissues and cell lines, whereas FGF9 mRNA was found underexpressed in LSCC tissues. MiR-372-3p directly bound to wild-type FGF9 mRNA 3'UTR, therefore led to the reduction in FGF9 expression. The upregulation of FGF9 or the downregulation of miR-372-3p substantially retarded LSCC cell growth, mitosis, and invasion. MiR-372-3p enhanced LSCC cell proliferation and invasion through inhibiting FGF9.
本研究旨在通过抑制 FGF9 研究 miR-372-3p 在肺鳞状细胞癌（LSCC）细胞增殖和侵袭中的作用。使用 RT-PCR 测定组织和细胞中 miR-372-3p 和 FGF9 mRNA 的表达。Western blot 用于测定组织和 NCI-H520 细胞系中 FGF9 的表达。双荧光素酶报告基因实验证实 FGF9 可被 miR-372-3p 直接靶向。MTT、集落形成实验用于研究外源性 miR-372-3p 和 FGF9 表达对 NCI-H520 细胞生长的影响。流式细胞术用于分析 miR-372-3p 和 FGF9 表达对细胞周期分布和凋亡的影响。Transwell 实验也用于观察 miR-372-3p 和 FGF9 表达对 NCI-H520 细胞侵袭能力的影响。发现 miR-372-3p 在 LSCC 组织和细胞系中均明显过表达，而 FGF9 mRNA 在 LSCC 组织中表达下调。miR-372-3p 可直接结合野生型 FGF9 mRNA 3'UTR，从而导致 FGF9 表达减少。FGF9 的上调或 miR-372-3p 的下调显著抑制 LSCC 细胞生长、有丝分裂和侵袭。miR-372-3p 通过抑制 FGF9 增强 LSCC 细胞的增殖和侵袭。
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