Advanced Biomedical Research

Micronucleus investigation in human buccal epithelial cells of gutkha users

Authors

1 Department of Zoology, Human Genetics and Toxicology Laboratory, Section of Genetics, Aligarh Muslim University, Aligarh, Uttar Pradesh, India

2 Department of Periodontics and Community Dentistry, Dr Z.A. Dental College, Aligarh Muslim University, Aligarh, Uttar Pradesh, India

Abstract

Background: Gutkha is a cheap and convenient betel quid substitute, which is popular among all age groups. Various studies reveal its carcinogenic nature that leads to oral submucosus fibrosis and increases the chances of oral cancer. The micronucleus (MN) assay in exfoliated mucosal cells is a useful method for observing genetic damage in humans.
Aim: To observe the genotoxic effect of gutkha on human buccal epithelial cells.
Materials and Methods: The MN assay was performed to assess the frequency of MN in human buccal epithelial cells. The study comprises 60 individuals of which 30 individuals were gutkha chewers and another 30 were nonusers (control). The MN frequency was scored to estimate the genotoxic damage.
Results: In gutkha users, the frequency of MN was highly significant (17.4 ± 0.944) as compared with nonusers (control) groups (4.53 ± 0.331) (P < 0.001).
Conclusions: The MN assay in human buccal epithelial cells is a useful and minimally invasive method for monitoring genetic damage in humans. Asignificantly higher frequency of micronucleated cells are found among gutkha users.

Keywords

1. IARC. Monographs on the Evaluation of Carcinogenic Risks of Chemicals to Humans, tobacco Habits Other than Smoking; Betel quid and Areca-nut Chewing; and Some Related Nitrosamines. Vol. 37. Lyon: IARC; 1985.  Back to cited text no. 1
    
2. IARC. Monographs on the Evaluation of Carcinogenic Risks to Humans, tobacco Habits Other than Smoking: Betel quid and Areca-nut Chewing. Vol. 85. Lyon: IARC; 2004.  Back to cited text no. 2
    
3. Kumar S. Panmasala chewing induces deterioration in oral health and its implications in carcinogenesis. Toxicol Mech Methods 2008;18:665-77.  Back to cited text no. 3
    
4. Tilakaratne WM, Klinikowski MF. Review on aetiology and pathogenesis. Oral Oncol 2005;42:561-8.  Back to cited text no. 4
    
5. Murti PR, Bhonsle RB, Pindborg JJ, Daftary DK, Gupta PC, Mehta FS. Malignant transformation rate in oral submucous fibrosis over a 17-year period. Community Dent Oral Epidemiol 1985;13:340-1.  Back to cited text no. 5
[PUBMED]    
6. Babu S, Sesikeran B, Bhat RV. Oral fibrosis among teenagers chewing tobacco, areca nut, and pan masala. Lancet 1996;348:692.  Back to cited text no. 6
[PUBMED]    
7. Gupta PC, Sinor PN, Bhonsle RB. Oral submucous fibrosis in India: A new epidemic? Natl Med J India 1998;11:113-6.  Back to cited text no. 7
    
8. Znaor A, Fuciæ A, Strnad M, Barkoviæ D, Škara M, Hozo I. Micronuclei in Peripheral Blood Lymphocytes as a Possible Cancer Risk Biomarker: A Cohort Study of Occupationally Exposed Workers in Croatia. Croat Med J 2003;44:441-6.  Back to cited text no. 8
    
9. Stich HF, Rosin MP. Quantitating the synergistic effect of smoking alcohol consumption with the micronucleus test on human buccal mucosa cells. Int J Cancer 1983;31:305-8.  Back to cited text no. 9
[PUBMED]    
10. Speit G, Schmid O. Local genotoxic effects of Formaldehyde in humans measured by the micronucleus test with exfoliated epithelial cells. Mutat Res 2006;613:1-9.  Back to cited text no. 10
[PUBMED]    
11. Stich HF, Stich W. Chromosome damaging activity of saliva of betel nut and tobacco chewers. Cancer Lett 1982;15:193-202.  Back to cited text no. 11
[PUBMED]    
12. Sundqvist K, Liu Y, Nair J, Bartsch H, Arvidson K, Grafstrom RC. Cytotoxic and genotoxic effects of areca nut-related compounds in cultured human buccal epithelial cells. Cancer Res 1989;49:294-8.  Back to cited text no. 12
    
13. Stich HF, Rosin MP. Micronuclei in exfoliated human cells as a tool for studies in cancer risk and cancer intervention. Cancer Lett 1984;22:241-53.  Back to cited text no. 13
[PUBMED]    
14. Stich HF, Dunn BP. DNA adducts, micronuclei and leukoplakias as intermediate endpoints in intervention trials. IARC Sci Publ 1988;89:137-45.  Back to cited text no. 14
[PUBMED]    
15. Surrallés J, Autio K, Nylund L, Jarventaus H, Norppa H, Veidebaum T, et al. Molecular cytogenetic analysis of buccal cells and lymphocytes from benzene-exposed workers. Carcinogenesis 1997;18:817-23.  Back to cited text no. 15
    
16. Rajeswari N, Ahuja YR, Malini U, Chandrashekar S, Balakrishna N, Rao KV, et al. Risk assessment in first-degree female relatives of breast cancer patients using the alkaline Comet assay. Carcinogenesis 2000;21:557-61.  Back to cited text no. 16
    
17. Heddle JA, Hite M, Kirkhart B, Mavournin K, MacGregor JT, Newell GW, et al. The induction of micronuclei as a measure of genotoxicity. A report of the U.S. Environmental Protection Agency Gene-Tox Program. Mutat Res 1983;123:61-118.  Back to cited text no. 17
    
18. Stich HF, Curtis JR, Parida BB. Application of the micronucleus test to exfoliated cells of high cancer risk groups: Tobacco chewers. Int J Cancer 1982;30:553-9.  Back to cited text no. 18
    
19. Roberts DM. Comparative cytology of the oral cavities of snuff users. Acta Cytol 1997;41:1008 -14.  Back to cited text no. 19
    
20. Ramirez A, Gattas GJF, Carvalho MB, Rapopport A, Saldanha PH. Clinical implications of micronuclei frequency as a biomonitor for alcoholic patients with oral carcinomas. Oral Oncol 1999;6:199-204.  Back to cited text no. 20
    
21. Nair U, Obe G, Nair J, Maru GB, Bhide SV, Pieper R, et al. Evaluation of frequency of micronucleated oral mucosa cells as a marker for genotoxic damage in chewers of betel quid with or without tobacco. Mutat Res 1991;261:163-8.  Back to cited text no. 21
    
22. Gandhi, Kaur R. Cytogenetic studies in exfoliated cells of high cancer risk groups, pan masala chewers. Hum Ecol 2000;9:221-8.  Back to cited text no. 22
    
23. Siddique YH, Ara G, Beg T, Afzal M. Micronucleus investigation in oral mucosal cells of gutkha/pan masala chewers. Indian Biologist 2008;40:1-5.  Back to cited text no. 23
    
24. Nair UJ, Nair J, Mathew B, Bartsch H. Glutathione S-transferasen M1 and T1 null genotypes as risk factors for oral leukoplakia in ethnic Indian betel quid/tobacco chewers. Carcinogenesis 1999;20:743-8.  Back to cited text no. 24
    
25. Wenke G, Rivenson A, Brunnemann KD, Hoffmann D, Bhide SV. A study of betel quid carcinogenesis. II. Formation of N nitrosamines during betel quid chewing. In: O'Neill IK, von Borstel RC, Miller CT, Long J, Bartsch H, editors. N-Nitroso Compounds: Occurrence, Biological Effects and Relevance to Human Cancer. Lyon: IARC: IARC Scientific Publications no. 57; 1984. p. 859-66.  Back to cited text no. 25
    
26. Murdia US, Mehta FJ, Bhide SV. Nitrate reductase activity and nitrite levels in the saliva of habitual users of various tobacco products. Food Chem Toxicol 1982;20:269-71.  Back to cited text no. 26
    
27. Nair J, Ohshima H, Pignatelli B, Friesen M, Calmels S, Bartsch H. Modifiers of endogenous carcinogen formation: Studies on in vivo nitrosation in tobacco users. In: Hoffmann D, Harris CC, editors. Mechanisms in Tobacco Carcinogenesis, Banbury Report 23. Cold Spring Harbour, NY: Cold Spring Harbour Laboratory Press; 1986. p. 45-61.  Back to cited text no. 27
    
28. Chakradeo PP, Nair J, Bhide SV. Endogenous formation of N-nitrosoproline and other N-nitrosamino acids in tobacco users. Cancer Lett 1994;86:187-94.  Back to cited text no. 28
    
29. Nair UJ, Floyd RA, Nair J, Bussachini V, Friesen M, Bartsch H. Formation of reactive oxygen species and of 8- hydroxydeoxyguanosine in DNA in vitro with betel quid ingredients. Chem Biol Interact 1987;63:157-69.  Back to cited text no. 29
    
30. Holland N, Bolognesi C, Kirsch-Volders M, Bonassi S, Zeiger E, Knasmueller S, et al. The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage: The HUMN project perspective on current status and knowledge gaps. Mutat Res 2008;659:93-108.  Back to cited text no. 30
    
31. Kamboj M, Mahajan S. Micronucleus- an upcoming marker of genotoxic damage. Clin Oral Investig 2007;11:121-6.  Back to cited text no. 31
    
32. Tolbert PE, Shy CM, Allen JW. Micronucleus and other nuclear anomalies in buccal smears: Methods development. Mutat Res 1992;271::69-77.  Back to cited text no. 32
    
33. Van Schooten FJ, Besaratinia A, De Flora S, Agostini FD, Izzotti A, Camoirano A, et al. Effects of oral administration of N-acetyl-L-cysteine: A multi-biomarker study in smokers. Cancer Epidemiol Biomarkers Prev 2002;11:167-75.  Back to cited text no. 33
    
Advanced Biomedical Research
Volume 2012, August
August 2012
Pages 1-4