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. 2017 Jun;50(3):e12336.
doi: 10.1111/cpr.12336. Epub 2017 Mar 20.

MicroRNA-802 plays a tumour suppressive role in tongue squamous cell carcinoma through directly _targeting MAP2K4

Xiaozhen Wu  1 , Zuode Gong  2 , Lanying Sun  3 , Long Ma  2 , Qibao Wang  2
Affiliations

MicroRNA-802 plays a tumour suppressive role in tongue squamous cell carcinoma through directly _targeting MAP2K4

Xiaozhen Wu et al. Cell Prolif. 2017 Jun.

Abstract

Objectives: Tongue squamous cell carcinoma (TSCC) is the most common oral tumours. MicroRNAs play crucial roles in many cell processes including cell viability, development, apoptosis, migration and invasion. The role of miR-802 in the TSCC is still unknown.

Materials and methods: The miR-802 expression in TSCC tissues and cell lines was determined by quantitative real-time polymerase chain reaction. CCK-8 assay was performed to measure the cell viability, while the cell invasion assay was used to determine the cell invasion. Dual-luciferase reporter and western blot were used to confirm the potential _target gene of miR-802.

Results: In our study, we demonstrated that miR-802 expression was downregulated in TSCC tissues and cell lines. Elevated expression of miR-802 suppressed the TSCC cell viability and invasion. Moreover, enforced expression of miR-802 increased the expression of E-cadherin, while suppressed the expression of N-cadherin, Snail and Vimentin in the TSCC cell. In addition, we identified the mitogen-activated protein kinase 4 (MAP2K4) as a direct _target gene of miR-802 in the TSCC cell. We also demonstrated that the expression of MAP2K4 was higher in the TSCC tissues than that in the adjacent normal tissues. Furthermore, the expression level of MAP2K4 was inversely associated with the expression of miR-802 in TSCC tissues. We also demonstrated that the MAP2K4 expression was upregulated in TSCC cell lines. Elevated expression of miR-802 inhibited TSCC cell viability and invasion through inhibiting MAP2K4 expression.

Conclusions: Our data revealed that miR-802 played as a tumour suppressor gene and might act as a therapeutic _target in TSCC patients.

Keywords: MAP2K4; miR-802; microRNAs; tongue squamous cell carcinoma.

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Figures

Figure 1
Figure 1
The expression of miR‐802 was decreased in tongue squamous cell carcinoma (TSCC) tissues. A, The expression of miR‐802 in 20 pairs of TSCC samples was shown. miR‐802 expression was detected by quantitative real‐time polymerase chain reaction. B, The miR‐802 expression level was downregulated in the 12 TSCC cases (12/20; 60%) compared to that in the adjacent normal tissues. C, The expression of miR‐802 was lower in the TSCC tissues than that in the adjacent normal tissues
Figure 2
Figure 2
Overexpression of miR‐802 suppressed the tongue squamous cell carcinoma (TSCC) cell viability. A, The miR‐802 expression in four TSCC cell lines (SCC1, SCC4, Cal27 and UM1) and normal oral keratinocyte cell culture (NHOK) and immortalized NOK16B cell line was determined by quantitative real‐time polymerase chain reaction. B, The expression level of miR‐802 was significantly increased in the SCC1 cell after treated with miR‐802 mimic. C, Elevated expression of miR‐802 suppressed SCC1 cell viability. D, Overexpression of miR‐802 decreased the ki‐67 expression in SCC1 cell. P<.01, and ***P<.001
Figure 3
Figure 3
Elevated expression of miR‐802 inhibited the TSCC cell invasion. A, Overexpression of miR‐802 suppressed SCC1 cell invasion. B, The relative invasive cells were shown. C, The expression of E‐cadherin was determined by quantitative real‐time polymerase chain reaction (qRT‐PCR). D, Overexpression of miR‐802 decreased the N‐cadherin expression. E, The expression of Snail was determined by qRT‐PCR. F, Elevated expression of miR‐802 suppressed the Vimentin expression. ***P<.001
Figure 4
Figure 4
MAP2K4 was a _target gene of miR‐802 in tongue squamous cell carcinoma (TSCC) cell and MAP2K4 expression was upregulated in TSCC tissues. A, The binding sites for miR‐802 within MAP2K4 were shown. B, The luciferase activity was decreased in SCC1 cell after treated with the miR‐802 mimic and MAP2K4‐WT plasmids. C, Overexpression of miR‐802 suppressed the MAP2K4 expression in SCC1 cell. D, The expression of MAP2K4 in 20 pairs of TSCC samples was shown. E, The MAP2K4 expression level was upregulated in 14 TSCC cases (14/20; 70%) compared to that in the adjacent normal tissues. F, The expression of MAP2K4 was higher in TSCC tissues than in the adjacent normal tissues. G, The expression level of MAP2K4 was inversely associated with miR‐802 in TSCC tissues
Figure 5
Figure 5
Elevated expression of miR‐802 inhibited tongue squamous cell carcinoma (TSCC) cell viability and invasion through _targeting MAP2K4. A, The MAP2K4 expression in four TSCC cell lines (SCC1, SCC4, Cal27 and UM1) and NHOK and NOK16B cell lines was determined by quantitative real‐time polymerase chain reaction (qRT‐PCR). B, The protein expression of MAP2K4 was detected by western blot. C, Overexpressed MAP2K4 promoted miR‐802‐transfected SCC1 cell viability. D, Overexpression of miR‐802 suppressed the TSCC cell invasion. E, The relative invasive cells were shown. F, The expression of E‐cadherin was determined by qRT‐PCR. G, Overexpression of MAP2K4 increased the N‐cadherin expression in the miR‐802‐transfected SCC1 cell. H, The expression of Snail was determined by qRT‐PCR. I, Elevated expression of MAP2K4 promoted the Vimentin expression in the miR‐802‐transfected SCC1 cell. *P<.05 and **P<.01
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