MiR-650 promotes proliferation and glycolysis of oral squamous cell carcinoma cells by targeting phosphoenolpyruvate carboxykinase 1
Objective:To analyze the mechanism of miR-650 on promoting proliferation and glycolysis of oral squamous cell carcinoma(OSCC)cells by targeting phosphoenolpyruvate carboxykinase 1(PCK1).Methods:The genes with differential expression and those related to cells proliferation and glycolysis were screened out by bioinformatics.CAL-27 and Tca8113 cell lines with stable overexpression PCK1 were constructed,and they were divided into Vector group and PCK1 group.OSCC cells with stable overexpression miR-650 were constructed,and they were divided into miR-NC group,miR-650 group and miR-650+PCK1 group.The cells proliferation ability in each group was detected by cell counting kit 8(CCK-8)and colony formation.The weight and volume of tumors were measured by tumor formation assay in nude mice.The expression level of PCK1 in tumor tissues was detected by immunohistochemistry.The expression level of PCK1 protein in each group was detected by Western blotting.The changes of glucose and lactic acid in cells were detected by kits of glucose and lactic acid.The database predicted PCK1 as the target and the dual-luciferase reporter experiment was performed to validate.The miR-650 in OSCC cells after transfection was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results:PCK1 was down-regulated in OSCC and was related to cells proliferation and glycolysis.Compared with the Vector group,the level of PCK1 protein was significantly increased in the PCK1 group.Overexpression of PCK1 could inhibit cells proliferation,inhibit growth of tumor volume and weight in nude mice.Overexpression of PCK1 could promote the production of glucose and inhibit its consumption,as well as inhibit the production of lactic acid.MiR-650 was up-regulated in OSCC compared with paracancerous tissues.Database prediction and dual-luciferase reporter assays confirmed the targeting relationship between miR-650 and PCK1.Compared with the miR-NC group,the expression levels of PCK1 mRNA and protein were significantly decreased in miR-650 group.The miR-650 could promote the proliferation of OSCC cells and regulate cells glycolysis by regulating the expression of PCK1.Conclusion:The miR-650 can promote the proliferation of OSCC cells and glycolysis by targeting PCK1.
万方数据
维普
