Porphyromonas gingivalis upregulates calbindin 1 and thus promotes the proliferation of gingival epithelial cells

doi:10.7518/hxkq.2022.01.014

Abstract

Abstract: Objective

This study aims to investigate the effect of calbindin 1 on the proliferation and apoptosis of gingival epithelial cells affected by Porphyromonas gingivalis (P. gingivalis)invitro.

Methods

A model of P. gingivalis infecting CA9-22 was established in vitro. At 24 h after infection, the expression of calbindin 1 (CALB1) was detected by real-time fluorescent quantitative polymerase chain reaction, Western blot, and immunofluorescence analyses. The expression of CALB1 was further inhibited by RNA interference. Cell proliferation was detected by BrdU analysis, and cell apoptosis was detected by caspase 3 activity. The expression of MDM2 and p53 was detected by Western blot analysis.

Results

P. gingivalis infection upregulated the expression of CALB1 in CA9-22 cells with multiplicity-dependent manner. CALB1 promoted the proliferation of CA9-22 cells, increased the expression of MDM2, and inhibited the expression of p53. Inhibiting CALB1 expression did not affect the inhibitory effect of P. gingivalis infection on CA9-22 apoptosis.

Conclusion

P. gingivalis infection can promote the proliferation of CA9-22 cells by increasing CALB1 expression. The related mechanism may be associated with MDM2-p53.

Key words: Porphyromonas gingivalis, gingival epithelial cell, cell proliferation, calbindin 1

CLC Number:

R 781.4

Zhang Yuwei, He Yuxuan, Ding Yi, Liu Chengcheng.. Porphyromonas gingivalis upregulates calbindin 1 and thus promotes the proliferation of gingival epithelial cells[J]. West China Journal of Stomatology, 2022, 40(1): 93-99.

Figures/Tables 4

References 28

1	Bagan J, Sarrion G, Jimenez Y. Oral cancer: clinical features[J]. Oral Oncol, 2010, 46(6): 414-417.
2	Markopoulos AK. Current aspects on oral squamous cell carcinoma[J]. Open Dent J, 2012, 6: 126-130.
3	Ganly I, Yang LY, Giese RA, et al. Periodontal pathogens are a risk factor of oral cavity squamous cell carcinoma, independent of tobacco and alcohol and human papillomavirus[J]. Int J Cancer, 2019, 145(3): 775-784.
4	Chocolatewala N, Chaturvedi P, Desale R. The role of bacteria in oral cancer[J]. Indian J Med Paediatr Oncol, 2010, 31(4): 126-131.
5	Katz J, Onate MD, Pauley KM, et al. Presence of Porphyromonas gingivalis in gingival squamous cell carcinoma[J]. Int J Oral Sci, 2011, 3(4): 209-215.
6	Fitzsimonds ZR, Rodriguez-Hernandez CJ, Bagaitkar J, et al. From beyond the pale to the pale riders: the emerging association of bacteria with oral cancer[J]. J Dent Res, 2020, 99(6): 604-612.
7	Schwaller B. The continuing disappearance of “pure” Ca²⁺ buffers[J]. Cell Mol Life Sci, 2009, 66(2): 275-300.
8	Liu Y, Porta A, Peng X, et al. Prevention of glucocorticoid-induced apoptosis in osteocytes and osteoblasts by calbindin-D28k[J]. J Bone Miner Res, 2004, 19(3): 479-
490
9	Bellido T, Huening M, Raval-Pandya M, et al. Calbindin-D28k is expressed in osteoblastic cells and suppresses their apoptosis by inhibiting caspase-3 activity[J]. J Biol Chem, 2000, 275(34): 26328-26332.
10	Sun S, Li F, Gao X, et al. Calbindin-D28K inhibits apoptosis in dopaminergic neurons by activation of the PI3-kinase-Akt signaling pathway[J]. Neuroscience, 2011, 199: 359-367.
11	Jin CJ, Lin T, Shan LQ. Downregulation of calbindin 1 by miR-454-3p suppresses cell proliferation in nonsmall cell lung cancer in vitro[J]. Cancer Biother Radiopharm, 2019, 34(2): 119-127.
12	Cao LQ, Wang YN, Liang M, et al. CALB1 enhances the interaction between p53 and MDM2, and inhibits the senescence of ovarian cancer cells[J]. Mol Med Rep, 2019, 19(6): 5097-5104.
13	Huang ZX, Fan GJ, Wang DL. Downregulation of calbindin 1, a calcium-binding protein, reduces the proliferation of osteosarcoma cells[J]. Oncol Lett, 2017, 13(5): 3727-3733.
14	Yang XY, Wang Y, Pang SL, et al. LINC00665 promotes the progression of acute myeloid leukemia by regulating the miR-4458/DOCK1 pathway[J]. Sci Rep, 2021, 11(1): 5009.
15	García-Cano J, Sánchez-Tena S, Sala-Gaston J, et al. Re-gulation of the MDM2-p53 pathway by the ubiquitin ligase HERC2[J]. Mol Oncol, 2020, 14(1): 69-86.
16	Jeon MH, Jeon CJ. Immunocytochemical localization of calretinin containing neurons in retina from rabbit, cat, and dog[J]. Neurosci Res, 1998, 32(1): 75-84.
17	Ozmen O, Topsakal S. Examination of skin lesions in rats with induced hyperthyroidism and hypothyroidism[J]. Biotech Histochem, 2020, 95(6): 438-444.
18	Korkmaz Y, Klinz FJ, Beikler T, et al. The Ca²⁺-binding protein calretinin is selectively enriched in a subpopulation of the epithelial rests of Malassez[J]. Cell Tissue Res, 2010, 342(3): 391-400.
19	Coleman H, Altini M, Ali H, et al. Use of calretinin in the differential diagnosis of unicystic ameloblastomas[J]. Histopathology, 2001, 38(4): 312-317.
20	Dorien FO, Roskams T, van den Eynde K, et al. The presence of endometrial cells in peritoneal fluid of women with and without endometriosis[J]. Reprod Sci, 2017, 24(2): 242-251.
21	Passebosc-Faure K, Li GR, Lambert C, et al. Evaluation of a panel of molecular markers for the diagnosis of malignant serous effusions[J]. Clin Cancer Res, 2005, 11(19Pt 1): 6862-6867.
22	Ohshima J, Wang Q, Fitzsimonds ZR, et al. Streptococcus gordonii programs epithelial cells to resist ZEB2 induction by Porphyromonas gingivalis[J]. Proc Natl Acad Sci U S A, 2019, 116(17): 8544-8553.
23	Arif SH. A Ca²⁺-binding protein with numerous roles and uses: parvalbumin in molecular biology and physiology[J]. Bioessays, 2009, 31(4): 410-421.
24	Tang X, Asano M, O'Reilly A, et al. P53 is an important regulator of CCL2 gene expression[J]. Curr Mol Med, 2012, 12(8): 929-943.
25	Kuboniwa M, Hasegawa Y, Mao S, et al. P. gingivalis accelerates gingival epithelial cell progression through the cell cycle[J]. Microbes Infect, 2008, 10(2): 122-128.
26	鲁泽, 潘亚萍. 牙龈卟啉单胞菌促进口腔鳞状细胞癌发生机制研究进展 [J]. 中国实用口腔科杂志, 2019, 12(10): 617-621.
	Lu Z, Pan YP. Research progress in the mechanism of Porphyromonas gingivalis in promoting oral squamous cell cancer[J]. Chin J Pract Stomatol, 2019, 12(10): 617-
621
27	Vousden KH. Outcomes of p53 activation—spoilt for choice[J]. J Cell Sci, 2006, 119(Pt 24): 5015-5020.
28	Oliner JD, Pietenpol JA, Thiagalingam S, et al. Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53[J]. Nature, 1993, 362(6423): 857-860.

[1]	Guo Lijia, Liu Yitong, Chen Yingyi, Xu Junji, Liu Yi. Liver macrophages show an immunotolerance phenotype in nonalcoholic fatty liver combined with Porphyromonas gingivalis-lipopolysaccharide infection [J]. West China Journal of Stomatology, 2023, 41(4): 385-394.
[2]	Zhang Min, Ao Xiaogang, Zheng Zheng, Chen Wenchuan.. Promoting the adhesion of human gingival epithelial cells on titanium surface by non-thermal atmospheric plasma irradiation [J]. West China Journal of Stomatology, 2022, 40(3): 285-292.
[3]	Zhang Xiujuan, Wu Wei, Chen Xin, Fang Zhichun, Ye Jinxiang, Ou Xiaoyan.. Effects of grape seed proanthocyanidins on the expression of inflammatory mediators in gingival epithelial cells [J]. West China Journal of Stomatology, 2022, 40(1): 39-44.
[4]	Jia Mei’e, Li Zhiyong, Xu Kai, Wang Yiheng, Yu Fei, He Xiangyi. Biological effects of exosome derived from Cal27 on normal human gingival fibroblasts [J]. West China Journal of Stomatology, 2021, 39(3): 313-319.
[5]	Wu Baoqin, Li Chunhui, Zhang Menglian, Nie Minhai. microRNA-1 gene delivery mediated by exosomes suppresses CAL-27 cell proliferation [J]. West China Journal of Stomatology, 2021, 39(2): 136-142.
[6]	Xia Xiaoyang, Fang Fei, Liu Yan, Che Chao, Ke Jinjuan, Jiang Shengjun. Expression of cyclophilin A in oral squamous cell carcinoma and its effect on cell proliferation and invasion [J]. West China Journal of Stomatology, 2021, 39(2): 164-169.
[7]	Yao Ke, Cai Jingyi, Zhao Lei, Wu Yafei, Zhao Zhihe, Shen Daonan. Research progress on two-component signal transduction systems in Porphyromonas gingivalis [J]. West China Journal of Stomatology, 2021, 39(1): 88-93.
[8]	Wang Liwei,Yu Yu,Chen Jiao,Feng Yun,Cui Bomiao,Li Xiaoying,Wang Jingnan,Chen Hongli,Zhang Ping. Protein kinase D1 regulates the growth and metabolism of oral squamous carcinoma cells in tumor microenvironment [J]. West China Journal of Stomatology, 2019, 37(6): 577-582.
[9]	Wang Chunmeng,Hong Lihua,Zhang Zhimin,Wang Yu. Research progress on the relationship between Porphyromonas gingivalis and the malignancy of the digestive system and possible pathogenetic mechanism [J]. West China Journal of Stomatology, 2019, 37(5): 521-526.
[10]	Yun Yang,Shanshan Chen,Chunmei Xu,Yafei Wu,Lei Zhao. Expression of triggering receptors expressed by myeloid cells-1 in macrophages stimulated by Porphyromonas gingivalis-lipopolysaccharide [J]. West China Journal of Stomatology, 2018, 36(5): 475-481.
[11]	Xuefeng Gao, Haibin Jiao, Changcheng Ye, Yingqun. Liu. Effects of protein kinase C and motigen-activated protein kinase kinase/extracellular regulated protein kinases signaling pathway on mRNA level of inducible nitric oxide synthase in Tca8113 cells [J]. West China Journal of Stomatology, 2018, 36(2): 133-139.
[12]	Chuanhua Li, Zhifeng Wang, Lina Zhu, Xin Fan, Jing Lan. Experimental research on Arginine-gingipain A gene vaccine from Porphyromonas gingivalis that prevents peri-implantitis in Beagle dogs [J]. West China Journal of Stomatology, 2018, 36(1): 76-81.
[13]	Yan Zhao, Yang Yu, Yurong Kou. Research advances on the molecular mechanism of autophagy regulated by Porphyromonas gingivalis [J]. West China Journal of Stomatology, 2017, 35(6): 654-658.
[14]	Jinmei Zhang, Lei Zhao, Yafei Wu. Research progress on the type Ⅸ secretion system of Porphyromonas gingivalis [J]. West China Journal of Stomatology, 2017, 35(5): 538-542.
[15]	Zhu Chen, Lingkai Su. Effects of paeonol on the function of bone marrow-derived macrophage from Porphyromonas gingivalis-induced mice [J]. West China Journal of Stomatology, 2017, 35(2): 139-144.