Estimation of Bcl-2 and Ki-67 in Gingival Epithelium of Epileptic Patients
Keywords:Gingival overgrowth, Bcl-2, Ki-67, Phenytoin
Introduction: Gingival overgrowth is one of several oral side effects of phenytoin, a potent antiepileptic drug. Several mechanisms have been elucidated to understand the pathogenesis of drug induced gingival overgrowth. The frequency of gingival overgrowth associated with chronic phenytoin therapy remains controversial. and the possible subclinical effects of this drug on the gingival epithelium should be investigated histopathologically and immunohistochemically.
Purpose of the study: To investigate the Bcl-2 for apoptosis rate and Ki-67 for the epithelial proliferative activity in epileptic patients.
Materials and methods: Twenty four samples of gingival tissue from epileptic patients treated with phenytoin and in eight samples of gingival tissue from healthy patients who didn’t use phenytoin (control) were evaluated for Bcl-2 and Ki-67 immunohistochemically.
Results: The results revealed moreproliferative activity of the overlying epithelium and an increased pattern of Bcl-2 and Ki-67 in phenytoin users compared to controls.
Conclusion: These results concluded that the increased epithelial thickness observed in phenytoin induced gingival overgrowth is associated with increased apoptotic rate and mitotic activity , especially in the oral epithelium.
Lin K, Guilhoto LM, Yacubian EM. Drug-induced gingival enlargement-Part II. Antiepileptic drugs: Not only phenytoin is involved. J Epilepsy Clin Neurophysiol 2007;13:83-8.
Wright G, Welbury RR, Hosey MT. Cyclosporin-induced gingival overgrowth in children. Int J Paediatr Dent 2005;15:403-11.
Boltchi FE, Rees TD, Iacopino AM. Cyclosporine a-induced gingival overgrowth: A comprehensive review. Quintessence Int 1999;30:775-83.
Euvard S, Kanitakis J, Claudy A. Skin cancers after organ transplantation. N Engl J Med 2003;348:1681-91.
Spolidorio LC, Spolidorio DM, Massucato EM, Neppelenbroek KH, Campanha NH, Sanches MH. Oral health in renal transplant recipients administered cyclosporin a or tacrolimus (FK506). Oral Dis 2006;12:309-14.
Cockfield SM. Identifying the patient at risk for post-transplant lymphoproliferative disorder. Transpl Infect Dis 2001;3:70-8.
Proctor R, Kumar N, Stein A, Moles D, Porter S. Oral and dental aspects of chronic renal failure. J Dent Res 2005;84:199-208.
Varga E, Tyldesley WR. Carcinoma arising in cyclosporin-induced gingival hyperplasia. Br Dent J 1991;171:26-7.
McLoughlin P, Newman L, Brown A. Oral squamous cell carcinoma arising in phenytoin-induced hyperplasia. Br Dent J 1995;178:183-4.
Penn I. Depressed immunity and the development of cancer. Cancer Detect Prev 1994;18:241-52.
Pramod S, Vijay K, Tanguturi S, Rachakonda H. Drug-induced gingival overgrowth: A critical insight into case reports from over two decades. J Indian Soc Periodontol 2016;20:496-502.
El Firt EY, Ghalwash DM. Ki-67 expression in gingival overgrowth: An immunohistochemical study. Life Sci J 2011;8:221-6.
Diop S, Letestu R, Orsolani D, Leboeuf Y, Le Tutour P, Thiam D, et al. Expression of proliferation marker Ki 67 in chronic lymphocytic leukemia. Dakar Med 2005;50:65-8.
Buduneli N, Buduneli E, Cxınar S, Lappin D, Kinane DF. Immunohistochemical evaluation of Ki-67 expression and apoptosis in cyclosporin a-induced gingival overgrowth. Periodontology 2007;78:282-9.
Kee N, Sivalingam S, Boonstra R, Wojtowicz JM. The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. J Neurosci Methods 2002;115:97-105.
Schluter C, Duchrow M, Wohlenberg C, Becker MH, Key G, Flad HD, Gerdes J. The cell proliferation-associated antigen of antobody Ki-67: A very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins. J Cell Biol 1993;123:513-22.
Saito K, Mori S, Tanda N, Sakamoto S. Expression of p53 protein and Ki-67 antigen I gingival hyperplasia induced by nifedipine and phenytoin. J Periodontol 1999;70:581-6.
Ananthanarayanan V, Deaton RJ, Yang XJ, Pins MR, Gann PH. Alteration of proliferation and apoptotic markers in normal and premalignant tissue associated with prostate cancer. BMC Cancer 2006;6:73.
Fulda S, Debatin KM. Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 2006;25:4798-811.
Jones ML. Connective tissue and stains. In: Bancroft JD, Gamble M, editors. Theory and Practice of Histological Techniques. 5th ed. Philadelphia, PA: Churchill Livingstone; 2002. p. 531-9.
Aimetti M, Romano F, Marsico A, Navone R. Non-surgical periodontal treatment of cyclosporin a-induced gingival overgrowth: Immunohistochemical results. Oral Dis 2008;14:244-50.
Kantarci A, Augustin P, Firatli E, Sheff MC, Hasturk H, Graves DT, et al. Apoptosis in gingival overgrowth tissues. J Dent Res 2007;86:888-92.
Abdollah M. Current opinion on drug-induced oral reactions: A comprehensive review. J Contemp Dent Pract 2008;9:1-15.
Schincaglia GP, Forniti F, Cavallini R, Piva R, Calura G, del Senno L. Cyclosporin-a increases Type I procollagen production and mRNA level in human gingival fibroblasts in vitro. J Oral Pathol Med 1992;21:181-5.
Li LL, Yin YZ, Wu ZK, Shi YJ. Expression of TGF-b1 and PCNA in cyclosporine A-induced gingival overgrowth. Shanghai Kou Qiang Yi Xue 2011;20:131-5.
Brunet L, Miranda J, Roset P, Berini L, Farre M, Mendieta C. Prevalence and risk of gingival enlargement in patients treated with anticonvulsant drugs. Eur J Clin Invest 2001;31:781-8.
Nurmenniemi PK, Pernu HE, Knuuttila ML. Mitotic activity of keratinocytes in nifedipine-and immunosuppressive medication-induced gingival overgrowth. J Periodontol 2001;72:167-73.
Das SJ, Newman HN, Olsen I. Keratinocyte growth factor receptor is up-regulated in cyclosporin a-induced gingival hyperplasia. J Dent Res 2002;81:683-7.
Myrillas TT, Linden GJ, Marley JJ, Irwin CR. Cyclosporin a regulates interleukin-1beta and interleukin-6 expression in gingiva: Implications for gingival overgrowth. J Periodontol 1999;70:294-300.
Tüter G, Serdar M, Yalim M, Gürhan IS, Balos K. Evaluation of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 levels in gingival fibroblasts of cyclosporin a-treated patients. J Periodontol 2002;73:1273-8.
Chong SA, Lee W, Arora PD, Laschinger C, Young EW, Simmons CA, et al. Methylglyoxal inhibits the binding step of collagen phagocytosis. J Biol Chem 2007;282:8510-20.
Humayun S, Prasad VR. Expression of p53 protein and Ki-67 antigen in oral premalignant lesions and oral squamous cell carcinomas: An immunohistochemical study. Natl J Maxillofac Surg 2011;2:38-46.
Vooijs GP. Benign proliferative reactions, intraepithelial neoplasia and invasive cancer of the uterine cervix. In: Bibbo B, editor. Comprehensive Cytopathology. Philadelphia, PA: W.B. Saunders; 1991. p. 153-230.
Yoshida T, Nagata J, Yamane A. Growth factors and proliferation of cultured rat gingival cells in response to cyclosporin a. J Periodont Res 2005;40:11-9.
Cetinkaya BO, Acikgoz G, Aydin O, Korkmaz A, CayirKeles G. The relationship between proliferating cell nuclear antigen expression and histomorphometrical alterations in cyclosporin a-induced gingival overgrowth in rats. Toxicol Pathol 2006;34:180-6.
Tu HP, Chen YT, Shieh YS, Chin YT Huang RY, Yang SF, et al. Cyclosporin-induced downregulation of the expression of E-cadherin during proliferation of edentulous gingival epithelium in rats. J Periodontol 2006;77:832-9.
Hyland PL, Mckeown ST, Mackenzie IC, Irwin CR. Regulation of keratinocyte growth factor and scatter factor in cyclosporin-induced gingival overgrowth. J Oral Pathol Med 2004;33:391-7.
Buduneli N, Sağol O, Atilla G, Duman S, Holmstrup P. Immunohistochemical analysis of epidermal growth factor receptor in cyclosporine induced-gingival overgrowth. Acta Odontol Scand 2001;59:367-71.
Bulut S, Ozdemir BH. Apoptosis and expression of caspase-3 in cyclosporin-induced gingival overgrowth. J Periodontol 2007;78:2364-8.
Willershausen-Zönnchen B, Lemmen C, Schumacher U. Influence of cyclosporine a on growth and extracellular matrix synthesis of human fibroblasts. J Cell Physiol 1992;152:397-402.
Cotrim P, Martelli-Junior H, Graner E, Sauk JJ, Coletta RD. Cyclosporin a induces proliferation in human gingival fibroblasts via induction of transforming growth factor-beta1. J Periodontol 2003;74:1625-33.
Deepa P, Visakan J. Cyclosporine a: Novel concepts in its role in drug-induced gingival overgrowth. Dent Res J (Isfahan) 2015;12:499-506.
Modéer T, Domeij H, Andurén I, Mustafa M, Brunius G. Effect of phenytoin on the production of interleukin-6 and interleukin-8 in human gingival fibroblasts. J Oral Pathol Med 2000;29:491-9.
Sato N, Matsumoto H, Akimoto Y, Fujii A. The effect of IL-1α and nifedipine on cell proliferation and DNA synthesis in cultured human gingival fibroblasts. J Oral Sci 2005;47:105-10.
Sakagami G, Sato E, Sugita Y, Kosaka T, Kubo K, Maeda H, et al. Effects of nifedipine and interleukin-1alpha on the expression of collagen, matrix metalloproteinase-1, and tissue inhibitor of metalloproteinase-1 in human gingival fibroblasts. J Periodontol Res 2006;41:266-72.
Nishikawa S, Nagata T, Morisaki I, Oka T, Ishida H. Pathogenesis of drug-induced gingival overgrowth. A review of studies in the rat model. J Periodontol 1996;67:463-71.
Niimi A, Tohnai I, Kaneda T, Takouchi M, Nagura H. Immunoistochemical analysis of effects of cyclosporin a on gingival epithelium. J Oral Pathol Med 1990;19:397-403.
TerHarmsel B, Smadts F, Kuupers J, Jeunnk M, Trimbost B, Ramaekers F. Bcl-2 immunoreactivity increases with severity of CIN: A study of normal cervical epithelia, CIN, and carcinoma. J Pathol 1996;197:26-30.
Jung JY, Jeong YJ, Jeong TS, Chung HJ, Kim WJ. Inhibition of apoptotic signals in overgrowth of human gingival fibroblasts by cyclosporin a treatment. Arch Oral Biol 2008;53:1042-9.
Bulut S, Ozdemir BH, Alaaddinoglu EE, Oduncuoglu FB, Bulut OE, Demirhan B. Effect of cyclosporin a on apoptosis and expression of p53 and Bcl-2 proteins in the gingiva of renal transplant patients. J Periodontol 2005;76:691-5.
Alaaddinoglu EE, Karabay G, Bulut S, Oduncuoglu FB, Ozdemir H, Bulut OE, et al. Apoptosis in cyclosporin a-induced gingival overgrowth: A histological study. J Periodontol 2005;76:166-70.
Birraux J, Kirby JA, Thomason JM, Taylor JJ. The effect of cyclosporin on cell division and apoptosis in human oral keratinocytes. J Periodont Res 2006;41:297-302.
Romito GA, Lotufo RF, Saraiva L, Pustiglioni AN, Pustiglioni FE, Stolf NA. Superinfecting microorganisms in patients under treatment with cyclosporin-a and its correlation to gingival overgrowth. Pesqui Odontol Bras 2003;17:35-40.
Strasser A, Harris AW, Bath ML, Cory S. Novel primitive lymphoid tumours induced in transgenic mice by cooperation between myc and bcl-2. Nature 1990;348:331-3.
Vaux DL, Cory S, Adams JM. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 1988;335:440-2.
Nakopoulou L, Vourlakou C, Zervas A, Tzonou A, Gakiopoulou H, Dimopoulos MA. The prevalence of Bcl-2, p53, and Ki-67 immunoreactivity in transitional cell bladder carcinomas and their clinicopathologic correlates. Hum Pathol 1998;29:146-54.
Maitu E, Sato M, Yamaki K. Effect of tranilast on martin metalloproteinase-1 secretion from human gingival fibroblast in vitro. J Periodontol 2004;75:1054-60.
Brunet L, Miranda J, Fere M, Berini L, Mendieta C. Gingival enlargement induced by drugs. Drug Saf 1996;15:219-31.
Gronzalez O, Gonzalez JM. Morphological and phenotypic difference in fibroblasts obtained from gingival overgrowth secondary to phenytoin: Pilot study. Rev Odontol Mex 2009;13:17-23.
Perucca E. An introduction to antiepileptic drugs. Epilepsia 2005;46:31-7.
Lucches J, Cortelli SH, Rodrigues J. Server phenytoin-induced gingival enlargement associated with periodontitis. Gen Dent 2008;56:199-203.
Reynolds EH. Chronic antiepileptic toxicity, a review. Epilepsia 1975;16:319-52.
Bhatia A, Prakash S. Topical phenytoin for wound healing. Dermatol Online J 2004;10:5.
Doufexi A, Mina M, Loannidou E. Gingival overgrowth in children: Epidemiology, pathogenesis and complications, a literature review. J Periodontol 2005;76:3-10.
Morisaki KK, Loyola-Rodriguez JP, Nagata T, Ishida H. Nifediphine-induced gingival overgrowth in the presence or absence of gingival inflammation in rats. J Period Res 1993;28:396-403.
Ishikawa S, Nagata T, Morisaki I, Oka T, Ishida H. Pathogenesis of drug induced gingival over-growth, a review of studies in the rat model. J Periodontol 1996;67:463-71.
Seymour RA, Heasann PA. Drugs and the periodontium. J Clin Periodontol 1992;19:1-11.
Hassel TM, Hefti AF. Drug induced gingival overgrowth: Old problem, new problem. Crit Rev Oral Biol Med 1991;2:103-7.
Barclay S, Thomason TM, Idle JR, Seymour RA. The incidence and severity of nifediphine induced gingival overgrowth. J Clin Periodontol 1992;19:311-4.
Seymour RA, Thomason JM, Ellis JS. The pathogenesis of drug induced gingival overgrowth. J Clin Periodontol 1996;23:165-75.
Easley J. Methods of determining alveolar osseous form. J Periodontol 1967;38:112.
Friedman N. Periodontal osseous surgery, osteoplasty and osteoectomy. J Periodontol 1955;26:257.
Goldman HM, Cohen DW. The infrabony pocket: Classification and treatment. J Periodontol 1958;29:272.
Ochsenbein C. A primer for osseous surgery. Int J Period Restor Dent 1986;6:9.
Carranza FA, Take H, Newman M. Clinical Periodontology. 10th ed. Philadelphia, PA: W.B. Saunders Co.; 2006. p. 18, 63, 270-2, 920-2.
Guncu GN, Caglayan F, Dincel A, Bozkurt A, Sayg S, Karabulut E. Plasma and gingival curricular fluid phenytoin concentrations as risk factors for gingival overgrowth. J Periodontol 2006;77:2005-10.
Seymour R. Effects of medications on the periodontal tissues in health and disease. J Periodontol 2006;40:120-9.
Yamada H, Nishimura F, Naruishi K, Chou H, Takashiba SH. Phenytoin and cyclosporine a suppress the expression of MMP-1, TTMP-1 and cathepsin, but not cathepsin B in cultured gingival fibroblasts. J Periodontol 2000;71:955-60.
Kataoka M, Kido J, Shinohara Y, Nagata T. Drug-induced gingival overgrowth-a review. Biol Pharm Bull 2005;28:1817-21.
Nurmenniemi PK, Pernu HE, Knuuttila ML. Mast cell subpopulations in gingival overgrowth induced by immunosuppressive and nifedipine medication. J Periodontol 2004;75:933-8.
Saito K, Mori S, Iwakura M, Sakamoto S. Immunohistochemical localization of transforming growth factor beta, basic fibroblast growth factor and heparansulphate glycosaminoglycan in gingival hyperplasia induced by nifedipine and phenytoin. J Period Res 1996;31:545-55.
Saygun I, Ozdemir A, Gunhan O, Aydintug YS, Karslioglu Y. Hereditary gingival fibromatosis and expression of Ki-67 antigen: A case report. J Periodontol 2003;74:873-8.
How to Cite
Asian Pacific Journal of Health Sciences applies the Creative Commons Attribution (CC-BY) license to published articles. Under this license, authors retain ownership of the copyright for their content, but they allow anyone to download, reuse, reprint, modify, distribute and/or copy the content as long as the original authors and source are cited. Appropriate attribution can be provided by simply citing the original article.