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, 24 (1), 841-849

MicroRNA-29b-3p Suppresses Oral Squamous Cell Carcinoma Cell Migration and Invasion via IL32/AKT Signalling Pathway

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MicroRNA-29b-3p Suppresses Oral Squamous Cell Carcinoma Cell Migration and Invasion via IL32/AKT Signalling Pathway

Jianya He et al. J Cell Mol Med.

Abstract

Oral squamous cell carcinoma (OSCC) is aggressive accompanied with poor prognosis. We previously isolated the most invasive cells resembling the invasive tumour front by microfluidic technology and explored their differentially expressed microRNAs (miRNAs) in our previous work. Here, we verified the miR-29b-3p as a guarder that suppressed migration and invasion of OSCC cells and was down-regulated in the most invasive cells. Besides that, the invasion suppression role of miR-29b-3p was achieved through the IL32/AKT pathway. Thus, miR-29b-3p and IL32 might serve as therapeutic targets for blocking the progression and improving the outcome of OSCC.

Keywords: AKT; IL32; OSCC; miR-29b-3p; migration and invasion.

Conflict of interest statement

The authors confirm that there are no conflicts of interest.

Figures

Figure 1
Figure 1
MiR‐29b‐3p showed low expression level in high invasive cell line UM‐SCC6‐M. A, Transwell invasion assay for UM‐SCC6 and UM‐SCC6‐M cells. B, Western blot of migration/invasion biomarkers' expression in UM‐SCC6 and UM‐SCC6‐M cells. C, qPCR analysis of RNA level of miR‐29b‐3p in UM‐SCC6 and UM‐SCC6‐M cells. Data represent the mean ± standard error of three independent experiments. *, P < .05; **, P < .01; ***, P < .001; ns, not significant
Figure 2
Figure 2
MiR‐29b‐3p suppressed OSCC cell migration and invasion. A, B, Effect of miR‐29b‐3p mimic on UM‐SCC6‐M cells' migration (A) and invasion (B), as evaluated with wound healing and transwell invasion assays, respectively. C, D, Effect of miR‐29b‐3p inhibitor on UM‐SCC6‐M cells' migration (C) and invasion (D), as evaluated with wound healing and transwell invasion assays, respectively. Scale Bar = 100 μm. Data represent the mean ± standard error of three independent experiments. *, P < .05; **P < .01; ***, P < .001; ns, P > .05. Ctrl, control; In, inhibitor; Mi, mimic
Figure 3
Figure 3
AKT signalling mediated the migration and invasion of OSCC cells. A, B, Effect of MK‐2206 on UM‐SCC6‐M cell migration (A) and invasion (B), as evaluated with wound healing and transwell invasion assays, respectively. C, D, Effect of AKT inhibitor MK‐2206 on the migration (C) and invasion (D) of UM‐SCC6 cells transfected with miR‐29b‐3p inhibitor as evaluated with wound healing and transwell invasion assays, respectively. Scale Bar = 100 μm. Data represent the mean ± standard error of three independent experiments. *, P < .05; **, P < .01; ***, P < .001; ns, not significant
Figure 4
Figure 4
IL32 was a downstream target of miR‐29b‐3p. A, Venn diagram shows the genes with high expression in UM‐SCC6‐M cells and down‐regulated by miR‐29b‐3p mimic. B, FPKM of IL32 in the mRNA sequencing data. C, D, mRNA (C) and protein (D) level of IL32 in UM‐SCC6 and UM‐SCC6‐M cells. E, Western blot showed the effects of miR‐29b‐3p mimic and inhibitor on IL32. Data represent the mean ± standard error of three independent experiments. ***, P < .001
Figure 5
Figure 5
IL32 promoted OSCC cell migration and invasion by activating AKT signalling. A, B, Effect of IL32 overexpression and knockdown on UM‐SCC6 cells' migration (A) and invasion (B), as evaluated with wound healing and transwell invasion assays, respectively. C, Effect of IL32 overexpression and knockdown on protein level of AKT and pAKT. D, E, Effect of AKT inhibitor MK‐2206 on the migration (D) and invasion (E) of UM‐SCC6 cells with IL32 overexpressed, as evaluated with wound healing and transwell invasion assays, respectively. Scale Bar = 100 μm. Data represent the mean ± standard error of three independent experiments. **, P < .01; ***, P < .001; KD, knockdown; ns, not significant; OE, overexpression
Figure 6
Figure 6
MiR‐29b‐3p suppressed migration and invasion via the IL32/AKT pathway. A, B, Effect of IL32 overexpression on the migration (A) and invasion (B) of UM‐SCC6‐M cells with miR‐29b‐3p mimic transfected, as evaluated with wound healing and transwell invasion assays, respectively. C, Western blot showed the AKT and pAKT level under the effect of IL32 and miR‐29b‐3p. Scale Bar = 100 μm. Data represent the mean ± standard error of three independent experiments. **, P < .01; ***, P < .001. Ctrl, control; In, inhibitor; KD, knockdown; Mi, mimic; OE, overexpression

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