Analysis of Cigarette-smoke-induced DNA Adducts by Butanol Extraction and Nuclease P1-enhanced 32P-postlabeling in Human Lymphocytes and Granulocytes

Overview

Journal Environ Health Perspect

Date 1993 Oct 1

PMID 8143607

Citations 2

Authors

K Savela

K Hemminki

Affiliations

Soon will be listed here.

Abstract

In an earlier study, we analyzed the aromatic DNA adducts separated from lymphocytes and granulocytes of smokers and nonsmokers using the nuclease P1-enhanced 32P-postlabeling assay. Here we compare the butanol extraction and nuclease P1-enhanced procedure on the same kind of samples. The DNA adducts of 42 per 10(8) nucleotides from smokers' lymphocytes were statistically higher (p < 0.05) than those of 11 from nonsmokers', when analyzed by the nuclease P1 treatment, but not by the 1-butanol extraction. The radioactivity obtained from the DNA digests on the TLC plates was lower in butanol-treated DNA samples when compared to those of nuclease P1 digestion. Lymphocytes appear to be a suitable test tissue for determining aromatic carcinogen exposure when detecting smoking-related DNA adducts by the nuclease P1-enhanced 32P-postlabeling analysis.

Citing Articles

Tracking matrix effects in the analysis of DNA adducts of polycyclic aromatic hydrocarbons.

Klaene J, Flarakos C, Glick J, Barret J, Zarbl H, Vouros P J Chromatogr A. 2015; 1439:112-123.

PMID: 26607319 PMC: 4789121. DOI: 10.1016/j.chroma.2015.10.057.

Exposure to urban and rural air pollution: DNA and protein adducts and effect of glutathione-S-transferase genotype on adduct levels.

Nielsen P, Okkels H, Sigsgaard T, Kyrtopoulos S, Autrup H Int Arch Occup Environ Health. 1996; 68(3):170-6.

PMID: 8919845 DOI: 10.1007/BF00381627.

References

Reddy M, Gupta R, RANDERATH E, Randerath K . 32P-postlabeling test for covalent DNA binding of chemicals in vivo: application to a variety of aromatic carcinogens and methylating agents. Carcinogenesis. 1984; 5(2):231-43. DOI: 10.1093/carcin/5.2.231. View

Perera F, Mayer J, Santella R, Brenner D, Jeffrey A, Latriano L . Biologic markers in risk assessment for environmental carcinogens. Environ Health Perspect. 1991; 90:247-54. PMC: 1519502. DOI: 10.1289/ehp.90-1519502. View

Everson R, RANDERATH E, Santella R, CEFALO R, Avitts T, Randerath K . Detection of smoking-related covalent DNA adducts in human placenta. Science. 1986; 231(4733):54-7. DOI: 10.1126/science.3941892. View

Stowers S, Anderson M . Formation and persistence of benzo(a)pyrene metabolite-DNA adducts. Environ Health Perspect. 1985; 62:31-9. PMC: 1568667. DOI: 10.1289/ehp.856231. View

Reddy M, Randerath K . Nuclease P1-mediated enhancement of sensitivity of 32P-postlabeling test for structurally diverse DNA adducts. Carcinogenesis. 1986; 7(9):1543-51. DOI: 10.1093/carcin/7.9.1543. View

Phillips D, Hemminki K, Alhonen A, Hewer A, Grover P . Monitoring occupational exposure to carcinogens: detection by 32P-postlabelling of aromatic DNA adducts in white blood cells from iron foundry workers. Mutat Res. 1988; 204(3):531-41. DOI: 10.1016/0165-1218(88)90047-x. View

Perera F, Hemminki K, Young T, Brenner D, Kelly G, Santella R . Detection of polycyclic aromatic hydrocarbon-DNA adducts in white blood cells of foundry workers. Cancer Res. 1988; 48(8):2288-91. View

Gupta R, Earley K, Sharma S . Use of human peripheral blood lymphocytes to measure DNA binding capacity of chemical carcinogens. Proc Natl Acad Sci U S A. 1988; 85(10):3513-7. PMC: 280242. DOI: 10.1073/pnas.85.10.3513. View

Phillips D, Hewer A, Martin C, Garner R, KING M . Correlation of DNA adduct levels in human lung with cigarette smoking. Nature. 1988; 336(6201):790-2. DOI: 10.1038/336790a0. View

10.

Gallagher J, Jackson M, George M, Lewtas J, Robertson I . Differences in detection of DNA adducts in the 32P-postlabelling assay after either 1-butanol extraction or nuclease P1 treatment. Cancer Lett. 1989; 45(1):7-12. DOI: 10.1016/0304-3835(89)90029-3. View

11.

Phillips D, Hewer A, Malcolm A, Ward P, Coleman D . Smoking and DNA damage in cervical cells. Lancet. 1990; 335(8686):417. DOI: 10.1016/0140-6736(90)90257-6. View

12.

Cuzick J, Routledge M, Jenkins D, Garner R . DNA adducts in different tissues of smokers and non-smokers. Int J Cancer. 1990; 45(4):673-8. DOI: 10.1002/ijc.2910450417. View

13.

Herbert R, Marcus M, Wolff M, Perera F, Andrews L, Godbold J . Detection of adducts of deoxyribonucleic acid in white blood cells of roofers by 32P-postlabeling. Relationship of adduct levels to measures of exposure to polycyclic aromatic hydrocarbons. Scand J Work Environ Health. 1990; 16(2):135-43. DOI: 10.5271/sjweh.1806. View

14.

Peluso M, Castegnaro M, Malaveille C, Talaska G, Vineis P, Kadlubar F . 32P-postlabelling analysis of DNA adducted with urinary mutagens from smokers of black tobacco. Carcinogenesis. 1990; 11(8):1307-11. DOI: 10.1093/carcin/11.8.1307. View

15.

Schooten F, Hillebrand M, van Leeuwen F, Lutgerink J, van Zandwijk N, Jansen H . Polycyclic aromatic hydrocarbon-DNA adducts in lung tissue from lung cancer patients. Carcinogenesis. 1990; 11(9):1677-81. DOI: 10.1093/carcin/11.9.1677. View

16.

Savela K, Hemminki K . DNA adducts in lymphocytes and granulocytes of smokers and nonsmokers detected by the 32P-postlabelling assay. Carcinogenesis. 1991; 12(3):503-8. DOI: 10.1093/carcin/12.3.503. View

17.

Schoket B, Phillips D, Hewer A, Vincze I . 32P-postlabelling detection of aromatic DNA adducts in peripheral blood lymphocytes from aluminium production plant workers. Mutat Res. 1991; 260(1):89-98. DOI: 10.1016/0165-1218(91)90084-y. View

18.

Gupta R . Enhanced sensitivity of 32P-postlabeling analysis of aromatic carcinogen:DNA adducts. Cancer Res. 1985; 45(11 Pt 2):5656-62. View