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Circadian rhythm variation of the clock genes Per1 and cell cycle related genes in different stages of carcinogenesis of buccal mucosa in animal model

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Author:
No author available
Journal Title:
Chinese Journal of Stomatology
Issue:
7
DOI:
10.3760/cma.j.issn.1002-0098.2015.07.003
Key Word:
基因表达;口腔肿瘤;癌,鳞状细胞;昼夜节律;细胞周期蛋白类;Gene expression;Mouth neoplasms;Carcinoma,squamous cell;Circadian rhythm;Cyclins

Abstract: Objective To investigate the expression and circadian rhythm variation of biological clock gene Per1 and cell cycle genes p53,CyclinD1,cyclin-dependent kinases (CDK1),CyclinB1 in different stages of carcinogenesis in buccal mucosa and its relationship with the development of buccal mucosa carcinoma.Methods Ninety golden hamsters were housed under 12 hours light-12 hours dark cycles,and the model of buccal squamous cell carcinoma was established by using the dimethylbenzanthracene(DMBA) to smear the golden hamster buccal mucosa.Before the DMBA was used and after DMBA was used 6 weeks and 14 weeks respectively,the golden hamsters were sacrificed at 6 different time points (5 rats per time point) within 24 hour,including 4,8,12,16,20 and 24 hour after lights onset(HALO),and the normal buccal mucosa,precancerous lesions and cancer tissue were obtained,respectively.HE stained sections were prepared to observe the canceration of each tissue.Real time RT-PCR was used to detect the mRNA expression of Per1,p53,CyclinD1,CDK1 and CyclinB1,and a cosine analysis method was applied to determine the circadian rhythm variation of Per1,p53,CyclinD1,CDK1 and CyclinB1 mRNA expression,which were characterized by median,amplitude and acrophase.Results The expression of Per1,p53,CDK1 and CyclinD1 mRNA in 6 different time points within 24 hours in the tissues of three different stages of carcinogenesis had circadian rhythm,respectively.However,the CyclinB 1 mRNA was expressed with circadian rhythm just in normal and cancer tissue (P<0.05),while in precancerous lesions the circadian rhythm was in disorder (P>0.05).As the development of carcinoma,the median of Per 1 and p53 mRNA expression were significantly decreased (P<0.05),yet the median of CDK1,CyclinB1 and CyclinD1 mRNA expression were significantly increased (P<0.05).The amplitude of Per1,p53 and CyclinD1 mRNA expression was significantly decreased as the development of carcinoma (P<0.05),however the amplitude of CDK1 mRNA expression was significantly increased (P<0.05).In addition,there was no significant difference in the amplitude of CyclinB1 mRNA expression.Tbe time that the peak expression value of Per1 and CDK1 mRNA appeared (Acrophase) in precancerous lesions was remarkably earlier than that in normal tissues,but the acrophase of p53 and CyclinD1 mRNA expression was remarkably delayed.Moreover,the acrophase of CDK1 and CyclinB1 mRNA expression in cancer tissues was obviously earlier than that in normal tissues,yet there was no significant variation in acrophase of Per1,p53,CyclinD1 mRNA expression between normal tissues and cancer tissues.Conclusions The circadian rhythm of clock gene Per1 and cell cycle genes p53,CyclinD1,CDK1,CyclinB1 expression remarkably varied with the occurrence and development of carcinoma.Further research into the interaction between circadian and cell cycle of two cycle activity and relationship with the carcinogenesis may provide new ideas and methods of individual treatment and the mechanism of carcinogenesis.

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