Regulating Effect Of Clock Gene PER1 On The Clock Gene Networks In Human Oral Squamous Cell Carcinoma

Objective: To investigate the effect of clock gene PER1 in oral squamous cell carcinoma cells on the expressions of other clock genes in clock gene networks.Methods: We used three groups PER1 short hairpin RNAs(shRNAs)to transfect SCC15 human oral squamous cell carcinoma cells.The best interference group was served as the experimental group with detection of western blot and quantitative real-time PCR(qRT-PCR).The negative control group was transfected scramble shRNA,which had no interference effect to any gene.The blank control group was SCC15 cells without making any treatment.We also used flow cytometry to detect proliferation and apoptosis level of the cell after PER1 knockdown,and qRT-PCR was used to detect the mRNA expressions of clock genes CLOCK,BMAL1,PER1,PER2,PER3,DEC1,DEC2,CRY1,CRY2,TIM,CKIε,RORα,NPAS2 and REV-ERBα in vitro and in vivo.Results: After PER1 knockdown in vitro and in vivo,the mRNA expressions of PER2,DEC1,DEC2,CRY1,CRY2 and NPAS2 were markedly decreased(P<0.05)and the mRNA expressions of PER3,TIM,RORα and REV-ERBα markedly were markedly increased(P<0.05).While mRNA expressions of CLOCK,BMAL1 and CKIε had no obvious alteration(P>0.05).After PER1 knockdown,apoptosis was decreased and cell proliferation and in vivo tumor formation were enhanced(P<0.05).Conclusion: Clock gene PER1 can regulate expressions of other clock genes in the clock gene networks,such as PER2,DEC1,DEC2,CRY1,CRY2,NPAS2,PER3,TIM,RORα and REV-ERBAα.PER1 gene plays an important role in regulating the clock gene networks.The role of PER1 in carcinogenesis is exerted not only by regulating downstream genes but also through the synergistic modulation of many other clock genes in the clock gene network.