|Table of Contents|

Biological clock gene PER1 regulates circadian rhythms of BAX, BCL2 and PCNA expression in oral squamous carcinoma cells



Research Field:
Publishing date:



Biological clock gene PER1 regulates circadian rhythms of BAX, BCL2 and PCNA expression in oral squamous carcinoma cells


YIN Shilin ZHANG Zhiwei TANG Hong YANG Kai

Department of Oral and Maxillofacial Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

PER1 circadian rhythm oral squamous cell carcinoma cell proliferation cell apoptosis

R394.3; R730.23; R739.85

ObjectiveTo explore the regulative role of biological clock gene PER1 in the circadian rhythms of proliferation, apoptosis and expression of intracellular BAX, BCL2 and PCNA in oral squamous carcinoma cells (OSCC). MethodsThe SCC15 cells stably overexpressing PER1 gene (OE-PER1-SCC15) and negative control SCC15 cells (NC-SCC15) infected with lentiviral virus without PER1 sequence were established. The TSCCA cells with stable PER1 knockdown (RNAi-PER1-TSCCA) and the negative control TSCCA cells (Scramble-TSCCA) of scramble plasmid lentivirus infection without PER1 fragment were also constructed. The cell proliferation index, apoptosis index and the expression of intracellular PER1, BAX, BCL2 and PCNA at 6 different time points within 24 h were detected in each group by flow cytometry, RT-qPCR and Western blotting, respectively. The data were subjected to ANOVA and cosine analysis, and the mesor, amplitude and acrophase were used as indicators to analyze the circadian rhythm characteristics of each gene expression. ResultsIn OE-PER1-SCC15 and NC-SCC15 cells, the cell proliferation index, apoptosis index, and the mRNA and protein levels of PER1, BAX, BCL2 and PCNA showed circadian rhythms (P<0.05). As compared with the NC-SCC15 cells, the OE-PER1-SCC15 cells had significantly increased mesors of apoptosis index and mRNA and protein levels of PER1 and BAX, while decreased mesors of proliferation index and mRNA and protein levels of BCL2 and PCNA (P<0.05). The amplitudes of apoptosis index and BAX mRNA level were remarkably elevated, while those of BCL2 and PCNA mRNA levels were reduced in OE-PER1-SCC15 cells (P<0.05). In addition, the acrophases of apoptosis index, and PER1 and BAX mRNA and protein levels were notably advanced, which were delayed in proliferation index, mRNA and protein levels of BCL2 and PCNA. When compared with the Scramble-TSCCA cells, the circadian rhythm of BAX protein expression was disturbed in the RNAi-PER1-TSCCA cells, but the cell proliferation index, apoptosis index and the mRNA and protein levels of PER1, BCL2 and PCNA, as well as mRNA level of BAX presented circadian rhythms (P<0.05), whose characteristics were opposite to those of OE-PER1-SCC15 cells. ConclusionThe changes in PER1 expression lead to the alteration in both its own circadian rhythm characteristics, and the circadian rhythm characteristics of proliferation, apoptosis, and BAX, BCL2, and PCNA expression in OSCC cells. 


[1]CHI A C, DAY T A, NEVILLE B W. Oral cavity and oropharyngeal squamous cell carcinoma: an update[J]. CA Cancer J Clin, 2015, 65(5): 401-421. DOI:10.3322/caac.21293. 
[2]CRAMER J D, BURTNESS B, LE Q T, et al. The changing therapeutic landscape of head and neck cancer[J]. Nat Rev Clin Oncol, 2019, 16(11): 669-683. DOI:10.1038/s41571-019-0227-z. 
[3]GIRALDI L, LEONCINI E, PASTORINO R, et al. Alcohol and cigarette consumption predict mortality in patients with head and neck cancer: a pooled analysis within the International Head and Neck Cancer Epidemiology (INHANCE) Consortium[J]. Ann Oncol, 2017, 28(11): 2843-2851. DOI:10.1093/annonc/mdx486. 
[4]DIBNER C, SCHIBLER U, ALBRECHT U. The mammalian circadian timing system: organization and coordination of central and peripheral clocks[J]. Annu Rev Physiol, 2010, 72: 517-549. DOI:10.1146/annurev-physiol-021909-135821. 
[5]TAKAHASHI J S. Transcriptional architecture of the mammalian circadian clock[J]. Nat Rev Genet, 2017, 18(3): 164-179. DOI:10.1038/nrg.2016.150. 
[6]KIM Y H, LAZAR M A. Transcriptional control of circadian rhythms and metabolism: a matter of time and space[J]. Endocr Rev, 2020, 41(5): 707-732. DOI:10.1210/endrev/bnaa014. 
[7]TUREK F W. Circadian clocks: not your grandfather’s clock[J]. Science, 2016, 354(6315): 992-993. DOI:10.1126/science.aal2613. 
[8]TAN X M, YE H, YANG K, et al. Circadian variations of clock gene Per2 and cell cycle genes in different stages of carcinogenesis in golden hamster buccal mucosa[J]. Sci Rep, 2015, 5: 9997. DOI:10.1038/srep09997. 
[9]GUAN D Y, XIONG Y, TRINH T M, et al. The hepatocyte clock and feeding control chronophysiology of multiple liver cell types[J]. Science, 2020, 369(6509): 1388-1394. DOI:10.1126/science.aba8984. 
[10]KORONOWSKI K B, SASSONE-CORSI P. Communicating clocks shape circadian homeostasis[J]. Science, 2021, 371(6530): eabd0951. DOI:10.1126/science.abd0951. 
[11]HANAHAN D, WEINBERG R A. Hallmarks of cancer: the next generation[J]. Cell, 2011, 144(5): 646-674. DOI:10.1016/j.cell.2011.02.013. 
[12]WEIGL Y, ASHKENAZI I E, PELEG L. Rhythmic profiles of cell cycle and circadian clock gene transcripts in mice: a possible association between two periodic systems[J]. J Exp Biol, 2013, 216(Pt 12): 2276-2282. DOI:10.1242/jeb.081729. 
[13]GRANDA T G, LIU X H, SMAALAND R, et al. Circadian regulation of cell cycle and apoptosis proteins in mouse bone marrow and tumor[J]. FASEB J, 2005, 19(2): 304-306. DOI:10.1096/fj.04-2665fje. 
[14]YANG G J, YANG Y X, TANG H, et al. Loss of the clock gene Per1 promotes oral squamous cell carcinoma progression via the AKT/mTOR pathway[J]. Cancer Sci, 2020, 111(5): 1542-1554. DOI:10.1111/cas.14362. 
[15]ZHAO N B, YANG K, YANG G L, et al. Aberrant expression of clock gene period1 and its correlations with the growth, proliferation and metastasis of buccal squamous cell carcinoma[J]. PLoS One, 2013, 8(2): e55894. DOI:10.1371/journal.pone.0055894. 
[16]GONG X B, TANG H, YANG K. PER1 suppresses glycolysis and cell proliferation in oral squamous cell carcinoma via the PER1/RACK1/PI3K signaling complex[J]. Cell Death Dis, 2021, 12(3): 276. DOI:10.1038/s41419-021-03563-5. 
[17]LIU Y, LANG H D, ZHOU M, et al. The preventive effects of pterostilbene on the exercise intolerance and circadian misalignment of mice subjected to sleep restriction[J]. Mol Nutr Food Res, 2020, 64(11): e1900991. DOI:10.1002/mnfr.201900991. 
[18]PARSONS R, PARSONS R, GARNER N, et al. CircaCompare: a method to estimate and statistically support differences in mesor, amplitude and phase, between circadian rhythms[J]. Bioinformatics, 2020, 36(4): 1208-1212. DOI:10.1093/bioinformatics/btz730. 
[19]FARSHADI E, VAN DER HORST G T J, CHAVES I. Molecular links between the circadian clock and the cell cycle[J]. J Mol Biol, 2020, 432(12): 3515-3524. DOI:10.1016/j.jmb.2020.04.003. 
[20]SULLI G, LAM M T Y, PANDA S. Interplay between circadian clock and cancer: new frontiers for cancer treatment[J]. Trends Cancer, 2019, 5(8): 475-494. DOI:10.1016/j.trecan.2019.07.002. 
[21]VOIGT R M, FORSYTH C B, KESHAVARZIAN A. Circadian rhythms: a regulator of gastrointestinal health and dysfunction[J]. Expert Rev Gastroenterol Hepatol, 2019, 13(5): 411-424. DOI:10.1080/17474124.2019.1595588. 
[22]DAVIS K, RODEN L C, LEANER V D, et al. The tumour suppressing role of the circadian clock[J]. IUBMB Life, 2019, 71(7): 771-780. DOI:10.1002/iub.2005. 
[23]BUCCITELLI C, SELBACH M. mRNAs, proteins and the emerging principles of gene expression control[J]. Nat Rev Genet, 2020, 21(10): 630-644. DOI:10.1038/s41576-020-0258-4. 
[24]杨凯, 郭伟, 孙沫逸, 等. 口腔鳞状细胞癌时辰化疗中国专家共识[J]. 中国口腔颌面外科杂志, 2019, 17(1): 7-12. DOI:10.19438/j.cjoms.2019.01.002. 
YANG K, GUO W, SUN M Y, et al. Chinese expert consensus on chrono-chemotherapy for oral squamous cell carcinoma[J]. China J Oral Maxillofac Surg, 2019, 17(1): 7-12. DOI:10.19438/j.cjoms.2019.01.002. 


Last Update: 2022-07-25