Keywords
Comet assay
DNA damage
Epithelial cells
Leukocytes
Mouth neoplasms.
DNA damage
Epithelial cells
Leukocytes
Mouth neoplasms.
How to Cite
1.
Rawat G, Urs AB, Chakravarti A, Kumar P. DNA damage in buccal cells in oral PMDs and malignant disorders by comet assay: a comparison with blood leukocytes. Braz. J. Oral Sci. [Internet]. 2019 Nov. 18 [cited 2024 Jul. 17];18:e191430. Available from: https://periodicos.sbu.unicamp.br/ojs/index.php/bjos/article/view/8657249
Abstract
Aim: DNA damage associated with Oral Squamous Cell Carcinoma (OSCC) and potentially malignant disorders (PMDs) is produced due to carcinogenic agents or increased oxidative stress. Comet assay can assist in early detection and evaluation of the amount of DNA damage; lymphocytesare the most commonly used cells for performing comet assay. Utilisation of buccal epithelial cells in comet assay can be a minimally invasive and rapid method. The present study compared the efficacy of comet assay in assessing DNA damage in buccal cells over peripheral blood leucocytes (PBLs) in oral potentially malignant and malignant disorders. Methods: The study included fifty five patients each of Leukoplakia, Oral Submucous Fibrosis (OSMF) and OSCC along with fifty five healthy individuals as control. Buccal epithelial cells were collected from all the selected subjects. DNA damage was evaluated bymeasuring the mean tail length (µm). Results: A significantly increased mean tail length (µm) and higher DNA damage were found in OSCC (26.1096 + 1.84355) and there was a progressive stepwise increase in mean tail length from control(8.4982 + 0.93307) to PMD [leukoplakia (14.6105 + 0.71857); OSMF (12.5009 + 1.12694)] to OSCC.The mean tail length in different habit groups was greater than controls, though no significant difference was noted between habit groups. The mean tail length of buccal cells was significantly greater than the mean tail length of PBLs in all study groups and controls. Conclusion: Hence, use of comet assay on buccal epithelial cells can prove to be beneficiary for evaluation of DNA damage.
References
1. Warnakulasuriya S, Johnson NW, Van der Waall I. Nomenclature and classification of potentially malignant disorders of oral mucosa. J Oral Pathol Med. 2007 Nov;36(10):575-80.
2. Yardimci G, Kutlubay Z, Engin B, Tuzun Y. Precancerous lesions of oral mucosa. World J Clin Cases. 2014 Dec 16;2(12):866-72. doi: 10.12998/wjcc.v2.i12.866.
3. KatarkarA, Mukherjee S, Khan M.H, Ray JG, Chaudhuri K. Comparative evaluation of genotoxicity by micronucleus assay in the buccal mucosa over comet assay in peripheral blood in oral precancer and cancer patients. Mutagenesis. 2014 Sep;29(5):325-34. doi: 10.1093/mutage/geu023.
4. Markopoulos AK. Current aspects on oral squamous cell carcinoma. Open Dent J. 2012;6:126-30. doi: 10.2174/1874210601206010126.
5. Choi S, Myers JN. Molecular pathogenesis of oral squamous cell carcinoma: implications for therapy. J Dent Res. 2008 Jan;87(1):14-32.
6. Carvalho AL, Singh B, Spiro RH, Kowalski LP, Shah JP. Cancer of the oral cavity: a comparison between institutions in a developing and a developed nation. Head Neck. 2004 Jan;26(1):31-8.
7. Mukherjee S, Ray JG, Chaudhuri K. Evaluation of DNA damage in oral precancerous and squamous cell carcinoma patients by single cell gel electrophoresis. Indian J Dent Res. 2011 Sep-Oct;22(5):735-6. doi: 10.4103/0970-9290.93475.
8. Jyoti S, Khan S, Naz F, Rahul, Ali F, Siddique YH. Assessment of DNA damage by panmasala, gutkha chewing and smoking in buccal epithelial cells using alkaline single cell gel electrophoresis (SCGE). Egypt J Med Hum Genet. 2013 Oct;14(4):391-4. doi: 10.1016/j.ejmhg.2013.07.004
9. Speit G, Witton-Davies T, Heepchantree W, Trenz K, Hoffmann H. Investigations on the effect of cigarette smoking in the comet assay. Mutat Res. 2003 Dec 9;542(1-2):33-42.
10. Tsai YS, Lee KW, Huang JL, Liu YS, Juo SH, Kuo WR, et al. Arecoline, a major alkaloid of areca nut, inhibits p53, represses DNA repair,and triggers DNA damage response in human epithelial cells. Toxicology. 2008 Jul 30;249(2-3):230-7. doi: 10.1016/j.tox.2008.05.007.
11. Rojas E, Valverde M, Sordo M, Ostrosky-Wegman P. DNA damage in exfoliated buccal cells of smokers assessed by the single cell gel electrophoresis assay. Mutat Res. 1996 Sep 13;370(2):115-20.
12. Rojas E, Lopez MC, Valverde M. Single cell gel electrophoresis assay: methodology and applications. J Chromatogr B Biomed Sci Appl. 1999 Feb 5;722(1-2):225-54.
13. Azqueta A, Collins AR. The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch Toxicol. 2013 Jun;87(6):949-68. doi: 10.1007/s00204-013-1070-0.
14. Monteith DK, Vanstone J. Comparison of the microgel electrophoresis assay and other assays for genotoxicity in the detection of DNA damage. Mutat Res. 1995 Dec;345(3-4):97-103.
15. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988 Mar;175(1):184-91.
16. Betti C, Davini T, Giannessi L, Loprieno N, Barale R. Comparative studies by comet test and SCE analysis in human lymphocytes from 200 healthy subjects. Mutat Res. 1995 Jul;343(4):201-7.
17. Faust F, Kassie F, Knasmüller S, Boedecker RH, Mann M, Mersch-Sundermann V. The use of the alkaline comet assay with lymphocytes in human biomonitoring studies. Mutat Res. 2004 May;566(3):209-29.
18. Collins AR. Investigating oxidative DNA damage and its repair using the comet assay. Mutat Res. 2009 Jan-Feb;681(1):24-32.
19. MøllerP, Knudsen LE, Loft S, Wallin H. The comet assay as a rapid test in biomonitoring occupational exposure to DNA-damaging agents and effect of confounding factors. Cancer Epidemiol Biomarkers Prev. 2000 Oct;9(10):1005-15.
20. Nandhakumar S, Parasuraman S, Shanmugam MM, Rao KR, Chand P, Bhat BV. Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay). J Pharmacol Pharmacother. 2011 Apr;2(2):107-11. doi: 10.4103/0976-500X.81903.
21. Cortés-Gutiérrez EI, Hernández-Garza F, García-Pérez JO, Dávila-Rodríguez MI, Aguado-Barrera ME, Cerda-Flores RM . Evaluation of DNA single and double strand breaks in women with cervical neoplasia based on alkaline and neutral comet assay techniques. J Biomed Biotechnol. 2012;2012:385245. doi: 10.1155/2012/385245.
22. Gupta RS, Gupta R, Goldstein S. Screening for genetic predisposition to mutagens in cancer patients. Exp Gerontol. 1996 Jan-Apr;31(1-2):267-80.
23. Udupa R, Hallikeri K, Trivedi DJ. The comet assay a method to measure DNA damage in oral submucous fibrosis patients: A case control study. Clin Cancer Invest. 2014;3(4):299-304.
24. Szeto YT, Benzie IF, Collins AR, Choi SW, Cheng CY, Yow CM, et al. A buccal cell model comet assay: development and evaluation for human biomonitoring and nutritional studies. Mutat Res. 2005 Oct 15;578(1-2):371-81.
25. Udumudi A, Jaiswal M, Rajeswari N, Desai N, Jain S, Balakrishna N, et al. Risk assessment in cervical dysplasia patients by single cell gel electrophoresis assay: a study of DNA damage and repair. Mutat Res. 1998 Jan 30;412(2):195-205.
26. Rawat G, Urs AB, Chakravarti A, Kumar P. Evaluation of DNA damage in peripheral blood leukocytes in oral potentially malignant and malignant disorders by comet assay. Clin Cancer Invest. 2018;7(2):50-5.
27. Liao W, McNutt MA, Zhu WG. The comet assay: a sensitive method for detecting DNA damage in individual cells. Methods. 2009 May;48(1):46-53. doi: 10.1016/j.ymeth.2009.02.016.
28. Guttikonda VR, Patil R, Kumar GS. DNA damage in peripheral blood leukocytes in tobacco users. J Oral Maxillofac Pathol. 2014 Sep;18(Suppl 1):S16-20. doi: 10.4103/0973-029X.141329.
29. Collins AR. Measuring oxidative damage to DNA and its repair with the comet assay. Biochim Biophys Acta. 2014 Feb;1840(2):794-800. doi: 10.1016/j.bbagen.2013.04.022.
30. Rajeswari N, Ahuja YR, Malini U, Chandrashekar S, Balakrishna N, Rao KV, et al. Risk assessment in first degree relatives of breast cancer patients using the alkaline comet assay. Carcinogenesis. 2000 Apr;21(4):557-61.
31. Zhang H, Buchholz TA, Hancock D, Spitz MR, Wu X. Gamma radiation induced single cell DNA damage as a measure of susceptibility to lung cancer: a preliminary report. Int J Oncol. 2000 Aug;17(2):399-404.
32. Baltaci V, Kayikçioğlu F, Alpas I, Zeyneloğlu H, Haberal A. Sister chromatid exchange rate and alkaline comet assay scores in patients with ovarian cancer. Gynecol Oncol. 2002 Jan;84(1):62-6.
33. Mohankumar MN, Janani S, Prabhu BK, Kumar PR, Jeevanram RK. DNA damage and integrity of UV-induced DNA repair in lymphocytes of smokers analysed by the comet assay. Mutat Res. 2002 Sep 26;520(1-2):179-87.
34. Jones NJ, Gregor AD, Waters R. Detection of DNA adducts in human oral tissue: Correlation of adduct levels with tobacco smoking and differential enhancement of adducts using the butanol extraction and nuclease P1 versions of 32p post labeling. Cancer Res. 1993 Apr 1;53(7):1522-8.
35. Dhawan A, Mathur N, Seth PK. The effect of smoking and eating habits on DNA damage in Indian population as measured in the Comet assay. Mutat Res. 2001 Mar 1;474(1-2):121-8.
36. Hoffmann H, Speit G. Assessment of DNA damage in peripheral blood of heavy smokers with the comet assay and the micronucleus test. Mutat Res. 2005 Mar 7;581(1-2):105-14.
37. Frenzilli G, Betti C, Davini T, Desideri M, Fornai E, Giannessi L et al. Evaluation of DNA damage in leukocytes of ex-smokers by single cell gel electrophoresis. Mutat Res. 1997 Apr 29;375(2):117-23.
2. Yardimci G, Kutlubay Z, Engin B, Tuzun Y. Precancerous lesions of oral mucosa. World J Clin Cases. 2014 Dec 16;2(12):866-72. doi: 10.12998/wjcc.v2.i12.866.
3. KatarkarA, Mukherjee S, Khan M.H, Ray JG, Chaudhuri K. Comparative evaluation of genotoxicity by micronucleus assay in the buccal mucosa over comet assay in peripheral blood in oral precancer and cancer patients. Mutagenesis. 2014 Sep;29(5):325-34. doi: 10.1093/mutage/geu023.
4. Markopoulos AK. Current aspects on oral squamous cell carcinoma. Open Dent J. 2012;6:126-30. doi: 10.2174/1874210601206010126.
5. Choi S, Myers JN. Molecular pathogenesis of oral squamous cell carcinoma: implications for therapy. J Dent Res. 2008 Jan;87(1):14-32.
6. Carvalho AL, Singh B, Spiro RH, Kowalski LP, Shah JP. Cancer of the oral cavity: a comparison between institutions in a developing and a developed nation. Head Neck. 2004 Jan;26(1):31-8.
7. Mukherjee S, Ray JG, Chaudhuri K. Evaluation of DNA damage in oral precancerous and squamous cell carcinoma patients by single cell gel electrophoresis. Indian J Dent Res. 2011 Sep-Oct;22(5):735-6. doi: 10.4103/0970-9290.93475.
8. Jyoti S, Khan S, Naz F, Rahul, Ali F, Siddique YH. Assessment of DNA damage by panmasala, gutkha chewing and smoking in buccal epithelial cells using alkaline single cell gel electrophoresis (SCGE). Egypt J Med Hum Genet. 2013 Oct;14(4):391-4. doi: 10.1016/j.ejmhg.2013.07.004
9. Speit G, Witton-Davies T, Heepchantree W, Trenz K, Hoffmann H. Investigations on the effect of cigarette smoking in the comet assay. Mutat Res. 2003 Dec 9;542(1-2):33-42.
10. Tsai YS, Lee KW, Huang JL, Liu YS, Juo SH, Kuo WR, et al. Arecoline, a major alkaloid of areca nut, inhibits p53, represses DNA repair,and triggers DNA damage response in human epithelial cells. Toxicology. 2008 Jul 30;249(2-3):230-7. doi: 10.1016/j.tox.2008.05.007.
11. Rojas E, Valverde M, Sordo M, Ostrosky-Wegman P. DNA damage in exfoliated buccal cells of smokers assessed by the single cell gel electrophoresis assay. Mutat Res. 1996 Sep 13;370(2):115-20.
12. Rojas E, Lopez MC, Valverde M. Single cell gel electrophoresis assay: methodology and applications. J Chromatogr B Biomed Sci Appl. 1999 Feb 5;722(1-2):225-54.
13. Azqueta A, Collins AR. The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch Toxicol. 2013 Jun;87(6):949-68. doi: 10.1007/s00204-013-1070-0.
14. Monteith DK, Vanstone J. Comparison of the microgel electrophoresis assay and other assays for genotoxicity in the detection of DNA damage. Mutat Res. 1995 Dec;345(3-4):97-103.
15. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988 Mar;175(1):184-91.
16. Betti C, Davini T, Giannessi L, Loprieno N, Barale R. Comparative studies by comet test and SCE analysis in human lymphocytes from 200 healthy subjects. Mutat Res. 1995 Jul;343(4):201-7.
17. Faust F, Kassie F, Knasmüller S, Boedecker RH, Mann M, Mersch-Sundermann V. The use of the alkaline comet assay with lymphocytes in human biomonitoring studies. Mutat Res. 2004 May;566(3):209-29.
18. Collins AR. Investigating oxidative DNA damage and its repair using the comet assay. Mutat Res. 2009 Jan-Feb;681(1):24-32.
19. MøllerP, Knudsen LE, Loft S, Wallin H. The comet assay as a rapid test in biomonitoring occupational exposure to DNA-damaging agents and effect of confounding factors. Cancer Epidemiol Biomarkers Prev. 2000 Oct;9(10):1005-15.
20. Nandhakumar S, Parasuraman S, Shanmugam MM, Rao KR, Chand P, Bhat BV. Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay). J Pharmacol Pharmacother. 2011 Apr;2(2):107-11. doi: 10.4103/0976-500X.81903.
21. Cortés-Gutiérrez EI, Hernández-Garza F, García-Pérez JO, Dávila-Rodríguez MI, Aguado-Barrera ME, Cerda-Flores RM . Evaluation of DNA single and double strand breaks in women with cervical neoplasia based on alkaline and neutral comet assay techniques. J Biomed Biotechnol. 2012;2012:385245. doi: 10.1155/2012/385245.
22. Gupta RS, Gupta R, Goldstein S. Screening for genetic predisposition to mutagens in cancer patients. Exp Gerontol. 1996 Jan-Apr;31(1-2):267-80.
23. Udupa R, Hallikeri K, Trivedi DJ. The comet assay a method to measure DNA damage in oral submucous fibrosis patients: A case control study. Clin Cancer Invest. 2014;3(4):299-304.
24. Szeto YT, Benzie IF, Collins AR, Choi SW, Cheng CY, Yow CM, et al. A buccal cell model comet assay: development and evaluation for human biomonitoring and nutritional studies. Mutat Res. 2005 Oct 15;578(1-2):371-81.
25. Udumudi A, Jaiswal M, Rajeswari N, Desai N, Jain S, Balakrishna N, et al. Risk assessment in cervical dysplasia patients by single cell gel electrophoresis assay: a study of DNA damage and repair. Mutat Res. 1998 Jan 30;412(2):195-205.
26. Rawat G, Urs AB, Chakravarti A, Kumar P. Evaluation of DNA damage in peripheral blood leukocytes in oral potentially malignant and malignant disorders by comet assay. Clin Cancer Invest. 2018;7(2):50-5.
27. Liao W, McNutt MA, Zhu WG. The comet assay: a sensitive method for detecting DNA damage in individual cells. Methods. 2009 May;48(1):46-53. doi: 10.1016/j.ymeth.2009.02.016.
28. Guttikonda VR, Patil R, Kumar GS. DNA damage in peripheral blood leukocytes in tobacco users. J Oral Maxillofac Pathol. 2014 Sep;18(Suppl 1):S16-20. doi: 10.4103/0973-029X.141329.
29. Collins AR. Measuring oxidative damage to DNA and its repair with the comet assay. Biochim Biophys Acta. 2014 Feb;1840(2):794-800. doi: 10.1016/j.bbagen.2013.04.022.
30. Rajeswari N, Ahuja YR, Malini U, Chandrashekar S, Balakrishna N, Rao KV, et al. Risk assessment in first degree relatives of breast cancer patients using the alkaline comet assay. Carcinogenesis. 2000 Apr;21(4):557-61.
31. Zhang H, Buchholz TA, Hancock D, Spitz MR, Wu X. Gamma radiation induced single cell DNA damage as a measure of susceptibility to lung cancer: a preliminary report. Int J Oncol. 2000 Aug;17(2):399-404.
32. Baltaci V, Kayikçioğlu F, Alpas I, Zeyneloğlu H, Haberal A. Sister chromatid exchange rate and alkaline comet assay scores in patients with ovarian cancer. Gynecol Oncol. 2002 Jan;84(1):62-6.
33. Mohankumar MN, Janani S, Prabhu BK, Kumar PR, Jeevanram RK. DNA damage and integrity of UV-induced DNA repair in lymphocytes of smokers analysed by the comet assay. Mutat Res. 2002 Sep 26;520(1-2):179-87.
34. Jones NJ, Gregor AD, Waters R. Detection of DNA adducts in human oral tissue: Correlation of adduct levels with tobacco smoking and differential enhancement of adducts using the butanol extraction and nuclease P1 versions of 32p post labeling. Cancer Res. 1993 Apr 1;53(7):1522-8.
35. Dhawan A, Mathur N, Seth PK. The effect of smoking and eating habits on DNA damage in Indian population as measured in the Comet assay. Mutat Res. 2001 Mar 1;474(1-2):121-8.
36. Hoffmann H, Speit G. Assessment of DNA damage in peripheral blood of heavy smokers with the comet assay and the micronucleus test. Mutat Res. 2005 Mar 7;581(1-2):105-14.
37. Frenzilli G, Betti C, Davini T, Desideri M, Fornai E, Giannessi L et al. Evaluation of DNA damage in leukocytes of ex-smokers by single cell gel electrophoresis. Mutat Res. 1997 Apr 29;375(2):117-23.
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