MicroRNA-140-5p inhibits the tumorigenesis of oral squamous cell carcinoma by targeting p21-activated kinase 4.

Abstract

Oral squamous cell carcinoma (OSCC) is a serious global health problem. Recently, accumulating microRNA (miRNA) has emerged as crucial players in the development and progression of carcinomas including OSCC. Our study aimed to further investigate the roles of miR-140-5p in OSCC tumorigenesis and related molecular basis. In this study, OSCC tissues and adjacent normal tissues were isolated from 34 OSCC patients who suffered from surgical resection at our hospital. MiR-140-5p level was measured by reverse-transcription quantitative polymerase chain reaction assay. p21-activated kinase 4 (PAK4) protein level was determined by western blot assay in OSCC cells at 48 h posttransfection or OSCC xenograft tumors at day 35 after OSCC cell injection. The cell proliferative ability was assessed by cell counting kit-8 assay in OSCC cells at 0, 24, 48, 72 h after transfection. Cell apoptosis and cell-cycle analysis was conducted using a flow cytometry in OSCC cells at 48 h after transfection. The interaction between miR-140-5p and PAK4 3'-untranslated region was tested by bioinformatics analysis and luciferase reporter assay in OSCC cells at 48 h after transfection. Mouse xenograft models of OSCC were established to examine the influence of miR-140-5p on OSCC tumorigenesis in vivo during 35 days after OSCC cell injection. Our data showed that miR-140-5p expression was notably downregulated in OSCC tissues and cell lines. MiR-140-5p inhibited the expression of PAK4 by direct interaction in OSCC cells. Functional analysis disclosed that miR-140-5p overexpression or PAK4 knockdown suppressed cell proliferation, promoted cell apoptosis, and induced cell-cycle arrest in OSCC. Moreover, PAK4 upregulation rescued the detrimental effects of miR-140-5p on cell proliferation and cell-cycle progression and hampered cell apoptosis induced by miR-140-5p in OSCC. In vivo experiments demonstrated that miR-140-5p overexpression suppressed the growth of OSCC xenograft tumors by downregulating PAK4. In conclusion, our data revealed miR-140-5p suppressed OSCC tumorigenesis by targeting PAK4 in vitro and in vivo, deepening our understanding on the function and molecular basis of miR-140-5p in the development of OSCC.