Effect of micro RNA-296-3p regulated cell on proliferation, migration and invasion of tongue squamous cell carcinoma by targeting nucleoprotein 1 / 中国医师进修杂志

ABSTRACT

Objective:To investigate the effects of micro RNA(miR)-296-3p on cell proliferation, migration and invasion of tongue squamous cell carcinoma and its underlying mechanism.Methods:Normal oral epithelial mucosal keratinocytes HOK was used as the control. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of miR-296-3p and nucleoprotein 1(NUPR1) mRNA in tongue squamous cell carcinoma cell lines CAL27, SCC15 and SCC9, and Western blot was used to detect the expression of NUPR1 protein. CAL27 cells were divided into NC group, miR-con group, miR-296-3p group, si-con group, si-NUPR1 group, miR-296-3p+pcDNA group and miR-296-3p+pcDNA-NUPR1 group, then CCK8 method was used to detect the cell activity. Transwell was used to detect the cell migration and invasion. Western blot was used to detect the protein expression levels of proliferating cell nuclear antigen (PCNA), matrixmetallo proteinase-2(MMP-2) and matrixmetallo proteinase-9(MMP-9). Dual-luciferase reporter assay system was implemented to verify the relationship between miR-296-3pand NUPR1.Results:Compared with the HOK cells, the content of miR-296-3p in the tongue squamous cell line CAL27, SCC15 and SCC9 groups was significantly reduced (0.54 ± 0.08, 0.38 ± 0.05, 0.59 ± 0.07 vs. 1.04 ± 0.12, t = 10.401, 15.231 , 9.718, P<0.05), while the expression levels of NUPR1mRNA (5.94 ± 0.40, 4.48 ± 0.45, 5.19 ± 0.48 vs.0.94 ± 0.12, t = 35.918, 22.803, 25.769) and protein (0.79 ± 0.09, 0.54 ± 0.05, 0.62 ± 0.08 vs. 0.28 ± 0.04, t = 15.535, 12.182, 11.404) were significantly increased (all P<0.05). Compared with those in the NC group or miR-con group, CAL27 cell activity, migration number, invasion number and the protein expression of PCNA, MMP-2 and MMP-9 in the miR-296-3p group were decreased (all P<0.05). Compared with those of the NC group or the si-con group, the CAL27 cell activity, migration number, number of invasions and the protein expression of PCNA, MMP-2 and MMP-9 in the si-NUPR1 group were decreased (all P<0.05). miR-296-3p negatively regulatedthe expression of NUPR1 in CAL27 cells. Compared with the miR-296-3p+pcDNA group, CAL27 cell viability (138.34 ± 5.73 vs. 98.54 ± 5.89, t = 14.530), migration number (95.28 ± 7.56 vs. 67.92 ± 5.23, t = 8.929), invasion number (184.53 ± 17.57 to 101.26 ± 10.64, t = 12.162) and the protein expression of PCNA (0.68 ± 0.07 to 0.35 ± 0.06, t = 10.738), MMP-2 (0.43 ± 0.05 to 0.29 ± 0.04, t = 6.559) and MMP-9 (0.58 ± 0.06 vs. 0.33 ± 0.08, t = 7.500) in the miR-296-3p+pcDNA-NUPR1 group were increased (all P<0.05). Conclusions:miR-296-3p may inhibitthe proliferation, migration and invasion of tongue squamous cell carcinoma cells by negatively regulating NUPR1.