关键词:

miR?498, 磷酸烯醇丙酮酸羧激酶 1, 口腔鳞状细胞癌, 细胞增殖, 细胞侵袭, 细 胞凋亡

Abstract:

Objective To investigate the effect of miR⁃498 targeting phosphoenolpyruvate carboxykinase 1 （PCK1）on growth of oral squamous cell carcinoma（OSCC）cells. Methods QRT⁃PCR was used to detect expression of miR⁃498 in tumor tissues from 150 OSCC patients and different OSCC cell lines HSC⁃3，SCC⁃15 and SCC⁃9. LipofectamineTM2000 transfection kit was used to transfect miR⁃498 mimics and anti⁃miR⁃498 into SCC⁃15 cells respectively. qRT⁃PCR was used to detect expression of miR⁃498 in the cells，so were Western blot to detect expression of PCK1 protein in the cells，CCK⁃8 to detect cell proliferation，flow cytometry to detect cell apoptosis， Transwell migration assay to detect cell migration and invasion，and dual ⁃luciferase reporter gene assay to detect the targeting relationship between miR⁃498 and PCK1. Co⁃transfection of miR⁃498 and PCK1 was used to further verify the effects of miR⁃498 and PCK1 on proliferation ，apoptosis，migration and invasion of SCC⁃15 cells. Results Compared with human normal oral keratinocyte HOK（1.03 ± 0.14），the expression level of miR⁃498 in human OSCC cell lines HSC⁃3（2.05 ± 0.21），SCC⁃15（2.75 ± 0.31），and SCC⁃9（2.31 ± 0.28）was significantly higher（F = 53.639，P < 0.001），and expression of miR⁃498 was the highest in SCC⁃15 cells. SCC⁃15 cells were then selected for the subsequent research. Compared with the control group and miR⁃NC group ，the number of migration and invasion of SCC⁃15 cells in miR⁃498 mimics group increased significantly（P < 0.05），and the apopto⁃ sis rate decreased significantly（P < 0.05）；compared with the control group and anti⁃miR⁃NC group，the number of migration and invasion of SCC⁃15 cells in anti⁃miR⁃498 group decreased significantly（P < 0.05），and the apoptosisrate increased significantly（P < 0.05）. The expression level of PCK1 protein in SCC⁃15 cells in the miR⁃498 mimics group（0.23 ± 0.02）was significantly lower than that the control group（0.51 ± 0.08）and miR⁃NC group（0.49 ± 0.06，P < 0.05）；The expression level of PCK1 protein in SCC⁃15 cells in the anti⁃miR⁃498 group（1.03 ± 0.05）was significantly higher than that in the control group（0.51 ± 0.08）and anti⁃miR⁃NC group（0.48 ± 0.07，P < 0.05）. miR ⁃498 negatively regulated PCK1 expression. Up ⁃ regulation of PCK1 expression reversed the effects of overex⁃ pression of miR⁃498 on proliferation，apoptosis，migration and invasion of SCC⁃15 cells. Conclusions MiR⁃498 can accelerate the proliferation，migration and invasion of SCC⁃15 cells and inhibit apoptosis of SCC⁃15 by inhibit⁃ ing the expression of PCK1，probably providing new molecular targets for the clinical diagnosis and treatment of OSCC

Key words: miR ?498,  , phosphoenolpyruvate carboxykinase 1,  , oral squamous cell carcinoma,  , cell proliferation,  , cell invasion,  , cell apoptosis