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. 2017 Jun;6(6):1323-1330.
doi: 10.1002/cam4.1026. Epub 2017 Apr 24.

MiR-372-3p promotes cell growth and metastasis by targeting FGF9 in lung squamous cell carcinoma

Qing Wang  1 , Siyang Liu  1 , Xitong Zhao  1 , Yuan Wang  1 , Dali Tian  1 , Wenjun Jiang  1
Affiliations

MiR-372-3p promotes cell growth and metastasis by targeting FGF9 in lung squamous cell carcinoma

Qing Wang et al. Cancer Med. 2017 Jun.

Abstract

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.

Keywords: LSCC; FGF9; MiR-372-3p.

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Figures

Figure 1
Figure 1
MiR‐372‐3p and FGF9 levels in tissues or cell lines. (A) The relative expression level of miR‐372‐3p in LSCC tissues and adjacent tissues. (B) The relative expression level of miR‐372‐3p in different LSCC cell lines. (C) The relative expression level of FGF9 mRNA in LSCC tissues and adjacent normal tissues. *P < 0.05.
Figure 2
Figure 2
MiR‐372‐3p binding to FGF9. (A) The complimentary sequences on miR‐372‐3p, wild‐type FGF9 3′UTR, and mutated type FGF9 3′UTR. (B) Relative luciferase activity of NCI‐H520 cells. (C) Relative expression of FGF9 in LSCC cells transfected with miR‐372‐3p inhibitors or NCs. *P < 0.05.
Figure 3
Figure 3
MiR‐372‐3p promoted LSCC cell growth and mobility. (A) The viability of NCI‐H520 in different groups. (B) The proliferation of NCI‐H520 cells in different groups. Flow cytometry results demonstrating cell cycle distribution (C) and apoptosis rates (D) of NCI‐H520 cells. (E) Transwell assay results showing LSCC the invasion of cells in different groups. All the experiments were individual and repeated for three times. *P < 0.05.
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