Effect of POLD1 on proliferation and invasion of oral squamous cell carcinoma and its regulatory mechanism

doi:10.19438/j.cjoms.2021.04.003

Abstract

Abstract: PURPOSE: To investigate the effect of POLD1 expression on cell proliferation, migration, invasion and cell cycle of oral squamous cell carcinoma (OSCC) cell lines and the relevant mechanism. METHODS: Online data were used to analyze POLD1 expression in head and neck squamous cell carcinoma (HNSCC) samples and normal tissues. POLD1 expression was evaluated in OSCC and normal tissues using immunohistochemistry. OSCC cell lines（SCC9 and CAL27) were infected with lentivirus, and the expression of POLD1 in stable cell lines was detected by Western blot. CCK-8 assay and EdU assay were used to detect cell proliferation; wound healing assay and Transwell migration and invasion assay were used to detect cell migration and invasion. The effect of POLD1 on cell cycle was detected by flow cytometry and the related pathway was studied via Western blot. SPSS 21.0 software package and GraphPad Prism were used for data analysis and plotting. RESULTS: POLD1 was abnormally upregulated in head and neck squamous cell carcinoma and OSCC tissues (P<0.001). Depletion of POLD1 in OSCC cells not only inhibited proliferation, migration, and invasion of cancer cells but also resulted in G2 arrest. The expression of P-Cdc2(Tyr15) and P-Rb(Ser807) in the experiment group were significantly increased. CONCLUSIONS: Down-regulation of POLD1 inhibits proliferation, migration and invasion of OSCC cells.

Key words: Oral squamous cell carcinoma, DNA polymerase delta subunit 1, Proliferation, Migration, Invasion, Cell cycle/G2 arrest, P-Cdc2

CLC Number:

R739.8

ZHANG Ying, WU Li, CHEN Yu-ling, LIN Yun-tao, SHEN Yue-hong, YANG Hong-yu. Effect of POLD1 on proliferation and invasion of oral squamous cell carcinoma and its regulatory mechanism[J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(4): 302-308.

References

[1] Chi AC, Day TA, Neville BW.Oral cavity and oropharyngeal squamous cell carcinoma--an update[J]. CA Cancer J Clin, 2015, 65(5): 401-421.
[2] Marur S, Forastiere A.Head and neck squamous cell carcinoma: update on epidemiology, diagnosis, and treatment[J].Mayo Clin Proc,2016, 91(3): 386-396.
[3] Almangush A, Mäkitie AA, Triantafyllou A, et al.Staging and grading of oral squamous cell carcinoma: an update[J]. Oral Oncol, 2020, 107: 104799.
[4] Vigneswaran N, Williams MD.Epidemiologic trends in head and neck cancer and aids in diagnosis[J]. Oral Maxillofac Surg Clin North Am, 2014, 26(2): 123-141.
[5] Angelis R, Sant M, Coleman M, et al.Cancer survival in Europe 1999-2007 by country and age: results of EUROCARE-5—a population-based study[J]. Lancet Oncol, 2014, 15(1): 23-34.
[6] Petersen PE.Strengthening the prevention of oral cancer: the WHO perspective[J]. Community Dent Oral Epidemiol, 2005, 33(6): 397-399.
[7] Nicolas E, Golemis EA, Arora S.POLD1: Central mediator of DNA replication and repair, and implication in cancer and other pathologies[J]. Gene, 2005, 590(1): 128-141.
[8] Byrnes JJ, Downey KM, Black VL, et al.A new mammalian DNA polymerase with 3' to 5'exonuclease activity: DNA polymerase delta[J]. Biochemistry, 1976, 15(13): 2817-2823.
[9] Johnson RE, Klassen R, Prakash L, et al.A major role of DNA polymerase δ in replication of both the leading and lagging DNA strands[J]. Mol Cell, 2015, 59(2): 163-175.
[10] Prindle MJ, Loeb LA.DNA polymerase delta in DNA replication and genome maintenance[J]. Environ Mol Mutagen, 2012, 53(9): 666-682.
[11] Lee MY, Zhang S, Lin SH, et al.Regulation of human DNA polymerase delta in the cellular responses to DNA damage[J]. Environ Mol Mutagen, 2012, 53(9): 683-698.
[12] Tahirov TH.Structure and function of eukaryotic DNA polymerase δ[J]. Subcell Biochem, 2012, 62: 217-236.
[13] Song J, Hong P, Liu C, et al.Human POLD1 modulates cell cycle progression and DNA damage repair[J]. BMC Biochem,2015,16:14-21.
[14] Kashkin K, Chernov I, Stukacheva E, et al.Cancer specificity of promoters of the genes controlling cell proliferation[J].J Cell Biochem, 2015, 116(2): 299-309.
[15] Qin Q,Tan Q, LiJ, et al. Elevated expression of POLD1 is associated with poor prognosis in breast cancer[J].Oncol Lett,2018,16(5): 5591-5598.
[16] Fang F, John W.Evidence that the Gl-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes[J]. Cell, 1991, 66(4): 731-742.
[17] Hu X,Moscinski LC.Cdc2: a monopotent or pluripotent CDK?[J]. Cell Prolif, 2011, 44(3): 205-211.
[18] Chea J,Zhang S, Zhao H, et al.Spatiotemporal recruitment of human DNA polymerase delta to sites of UV damage[J]. Cell Cycle, 2012, 11(15): 2885-2895.
[19] Gould KL,Nurse P.Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis[J]. Nature, 1989, 342(6245): 39-45.
[20] Norbury C, Nurse P.Animal cell cycles and their control[J]. Annu Rev Biochem, 1992, 61: 441-470.
[21] Solomon MJ, Lee T, Kirschner MW.Role of phosphorylation in p34cdc2 activation: identification of an activating kinase[J]. Mol Biol Cell, 1992, 3(1): 13-27.
[22] Wells NJ, Watanabe N, Tokusumi T, et al.The C-terminal domain of the Cdc2 inhibitory kinase Myt1 interacts with Cdc2 complexes and is required for inhibition of G(2)/M progression[J]. J Cell Sci, 1999, 112(Pt 19): 3361-3371.
[23] Herrera RE, Mäkelä TP, Weinberg RA.TGF beta-induced growth inhibition in primary fibroblasts requires the retinoblastoma protein[J]. Mol Biol Cell, 1996, 7(9): 1335-1342.
[24] Herrera RE, Sah VP, Williams BO, et al.Altered cell-cycle kinetics, gene expression,and G1 restriction point regulation in Rb-deficient fibroblasts[J]. Mol Cell Biol,1996,16(3): 2402-2407.
[25] Sherr CJ.Cancer cell cycles[J]. Science, 1996, 274(5293): 1672-1677.