140 A.H.H.
Letters
to
INDUCIBILITY, SMOKING HABITS,
AND AGE AT DIAGNOSIS OF
LARYNGEAL CARCINOMA
the Editor
ARYL HYDROCARBON HYDROXYLASE INDUCIBILITY AND LARYNGEAL CARCINOMAS
SIR,-Smokers are predisposed to cancer of the lung, bladder, and mouth and larynx.1-3 The cancer risk increases with the amount of tobacco smoke inhaled. Genetic differences in
susceptibility
to
smoke
carcinogens have, however, been
demonstrated.4 The polycyclic aromatic hydrocarbons (P.A.H.), regarded as the main carcinogens of tobacco smoke, are metabolically activated in the cells. This activation is mediated by aryl hydrocarbon hydroxylase (A.H.H.),5 6 hydroxylating the P.A.H. to epoxides, which form covalent bonds with D.N.A., R.N.A., and certain proteins.78 It has been suggested that this epoxide-induced D.N.A. alteration might represent a biochemical basis of
carcinogenesis.9 is inducible by various’agents.10 In man, A.H.H. inducibility has been assayed in leucocytes, especially lymphocytes, foreskin tissue cultures, and pulmonary alveolar macrophages.1l-14 Fibroblasts from different organs in the same embryo metabolise P.A.H. to the same extent, while there are variations in the metabolism of P.A.H. in fibroblasts from different embryos. 15 This genetic heterogeneity of A.H.H. inducibility in man has been extensively studied by Kellerman et al., who suggested a single autosomal locus of genetic control, with three different phenotypes of low, intermediate, and high inducibility,16 representing 44·7%, 45-9%, and 9.4% of the population, respectively.l’ These frequencies accord with preliminary data from Sweden.18 19 Kellerman et al. reported a significantly increased incidence of bronchogenic carcinomas in the group with high A.H.H. inducibility.1’ This also raised the question of an association between A.H.H. activity and oral and laryngeal carcinomas in smokers,2O upon which we now present some preliminary data. 65 patients (64 males, 1 female) with invasive laryngeal carcinoma were diagnosed in Malmö, Sweden, in the period 1962-75. The patients had all undergone surgery and/or radiotherapy, and previous histopathological examinations had shown all carcinomas to be of the spinocellular type, mainly highly differentiated. In December, 1975, 55 patients (all males) were still alive, and all who were below 80 years of age (45) were invited to the section of preventive medicine, Malmö General Hospital, A.H.H.
1. 2.
Doll, R., Hill, A. B. Br. med. J. 1950, ii, 739. Wynder, E. L., Mabuchi, K. in Advances in Tumour Prevention, Detection and Characterization (edited by C. Maltoni); vol. II p. 146. Amsterdam,
1974. 3. Hammond, E. C., Selikoff, I. J., Seideman, H. in Cancer Epidemiology: Environmental Factors (edited by P. Bucalossi, U. Veronesi, and N. Cascinelli); vol. III, p. 147. Amsterdam, 1975. 4. Tokuhata, G. K. Am. J. Popul. Hlth, 1964, 54, 24. 5. Gelboin, H. V., Kinoshita, N., Wiebel, F. J. Fed Proc. 1972, 31, 1298. 6. Huberman, E., Aspiras, L., Heidelberger, C., Grover, P. L., Sims, P. Proc. natn Acad. Sci. U.S.A 1971, 68, 3195. 7. Grover, P. L., Sims, P. Biochem. Pharmac. 1973, 22, 661. 8. Rogan, E. G., Cavalieri, E. Biochem. Biophys. Res. Commun. 1974, 58, 1119. 9. Love, D. L. Naval Ordnance Laboratory technical report no. 74-197, Nov.
8, 1974. 10. Conney, A. H. Pharmac. Rev. 1967, 19, 317. 11. Busbee, D. L., Shaw, C. R., Cantrell, E. T. Science, 1972, 178, 315. 12. Whitlock, J. P. Jr., Cooper, H. L., Gelboin, H. L. ibid. 1972, 177, 618. 13. Levin, W., Conney, A. H., Alvares, A. P., Merkatz, I., Kappas, A. ibid. 1972, 176, 419. 14. Cantrell, E. T., Warr, G. A., Busbee, D. L., Martin, R. R. J. clin. Invest. 1973, 52, 1881. 15. Huberman, E., Sachs, L. Inc. J. Cancer, 1973, 11, 412. 16. Kellerman, G., Luyten-Kellerman, M., Shaw, C. R. Am. J. hum. Genet. 1973, 25, 327. 17. Kellerman, G., Shaw, C. R., Luyten-Kellerman, M. New Engl. J Med. 1973, 289, 934. 18. Löfroth, G., Natarajan, A. T. Läkartidn. 1975, 72, 593. 19. Korsgaard, R., and others. Unpublished. 20. Lancet, 1974, i, 910.
’Based upon 15 complete smoking histories. tBased upon 13 complete smoking histories; 1 non-smoker in this group. based upon 7 complete smoking histories; 1 non-smoker in this group. Classification in high, intermediate and low A.H.H. inducibility groups, and calculation of the expected numbers of patients in each group is according to Keller17 man et al.
for A.H.H. assessment. 38 patients left venous blood-samples which were assayed for A.H.H. inducibility at the department of tumour cytogenetics, Lund. Lymphocyte suspensions were prepared by a one-step centrifugal technique using ’Lymphoprep’, and the lymphocytes were then cultured in Parker 199 medium complemented with 15% fetal bovine serum, glutamine, antibiotics, and phytohmmagglutinin. After incubation at 37°C for 72 h 1-5 5 mmol/1 3-methylcholanthrene in acetone was added and allowed to take effect for an additional 24 h. The A.H.H. inducibility was then assayed by a modification of the method of Nebert and Gelboin.21 A high correlation between inducible A.H.H. activity in different cells from the same individuals has been demonstrated in previous studies.22 Our results are presented in the table. These preliminary data support the concept that A.H.H. plays an important role in the activation ofp.A.H. to ultimate carcinogens. The statisti-
cally highly significant (by chi-square test) over-representation of patients with high A.H.H. inducibility in this group of carcinomas might be of some value in identifying individuals at high risk. Mortality-rates of carcinomas of the respiratory tract are increasing, and screening of A.H.H. inducibilÜy23 24 in addition to sputum cytology, chest X-rays, and so on might improve survival figures, and even be of value in preventive medicine. This study was supported by grants from the Swedish National Association against Heart and Chest Diseases. R. K. is the recipient of a personal research grant (746-B74-02P) from the Swedish Cancer Socictv. Section of Preventive Medicine, Department of Internal Medicine and Department of Oto-rhino-laryngology, University of Lund, Malmö General Hospital, Malmö, Sweden; Research Department I and Department of Lung Medicine, University Hospital, Lund; and Department of Tumour
Cytogenetics, Wallenberg Laboratory, University of Lund
E. R. B. P.
TRELL
KORSGAARD HOOD KITZING GUNNELA NORDÉN B. G. SIMONSSON
COFFEE, MYOCARDIAL INFARCTION, AND SUDDEN DEATH
laboratory2 may shed light on the continuing controversy about the relation between coffee consumption and the incidence of myocardial infarction (M.l.) and subsequent death. Prospective studies26 27 have shown no corSIR,-Work in
our
21. Nebert, D. W., Gelbom, H. J. biol. Chem. 1968, 243, 6242. 22. Cantrell, E., Busbee, D., Warr, G., Martin, R. Life Sci. 1973, 13, 1649. 23. Chest, 1975, 67, 508. 24. Brooks, S. M. J. occup. Med. 1975, 17, 19. 25. Rotenberg, F. A., DeFeo, J. J. Fed Proc. 1976, 35, 384. 26. Dawber, T. R., Kannel, W. B., Gordon, T. New Engl. J. Med. 1974, 291, 871. 27. Klatsky, A. L., Friedman, G. D., Siegelaub, A. B. J. Am. med. Ass. 1973,
226, 540.
Letters
to
INDUCIBILITY, SMOKING HABITS,
AND AGE AT DIAGNOSIS OF
LARYNGEAL CARCINOMA
the Editor
ARYL HYDROCARBON HYDROXYLASE INDUCIBILITY AND LARYNGEAL CARCINOMAS
SIR,-Smokers are predisposed to cancer of the lung, bladder, and mouth and larynx.1-3 The cancer risk increases with the amount of tobacco smoke inhaled. Genetic differences in
susceptibility
to
smoke
carcinogens have, however, been
demonstrated.4 The polycyclic aromatic hydrocarbons (P.A.H.), regarded as the main carcinogens of tobacco smoke, are metabolically activated in the cells. This activation is mediated by aryl hydrocarbon hydroxylase (A.H.H.),5 6 hydroxylating the P.A.H. to epoxides, which form covalent bonds with D.N.A., R.N.A., and certain proteins.78 It has been suggested that this epoxide-induced D.N.A. alteration might represent a biochemical basis of
carcinogenesis.9 is inducible by various’agents.10 In man, A.H.H. inducibility has been assayed in leucocytes, especially lymphocytes, foreskin tissue cultures, and pulmonary alveolar macrophages.1l-14 Fibroblasts from different organs in the same embryo metabolise P.A.H. to the same extent, while there are variations in the metabolism of P.A.H. in fibroblasts from different embryos. 15 This genetic heterogeneity of A.H.H. inducibility in man has been extensively studied by Kellerman et al., who suggested a single autosomal locus of genetic control, with three different phenotypes of low, intermediate, and high inducibility,16 representing 44·7%, 45-9%, and 9.4% of the population, respectively.l’ These frequencies accord with preliminary data from Sweden.18 19 Kellerman et al. reported a significantly increased incidence of bronchogenic carcinomas in the group with high A.H.H. inducibility.1’ This also raised the question of an association between A.H.H. activity and oral and laryngeal carcinomas in smokers,2O upon which we now present some preliminary data. 65 patients (64 males, 1 female) with invasive laryngeal carcinoma were diagnosed in Malmö, Sweden, in the period 1962-75. The patients had all undergone surgery and/or radiotherapy, and previous histopathological examinations had shown all carcinomas to be of the spinocellular type, mainly highly differentiated. In December, 1975, 55 patients (all males) were still alive, and all who were below 80 years of age (45) were invited to the section of preventive medicine, Malmö General Hospital, A.H.H.
1. 2.
Doll, R., Hill, A. B. Br. med. J. 1950, ii, 739. Wynder, E. L., Mabuchi, K. in Advances in Tumour Prevention, Detection and Characterization (edited by C. Maltoni); vol. II p. 146. Amsterdam,
1974. 3. Hammond, E. C., Selikoff, I. J., Seideman, H. in Cancer Epidemiology: Environmental Factors (edited by P. Bucalossi, U. Veronesi, and N. Cascinelli); vol. III, p. 147. Amsterdam, 1975. 4. Tokuhata, G. K. Am. J. Popul. Hlth, 1964, 54, 24. 5. Gelboin, H. V., Kinoshita, N., Wiebel, F. J. Fed Proc. 1972, 31, 1298. 6. Huberman, E., Aspiras, L., Heidelberger, C., Grover, P. L., Sims, P. Proc. natn Acad. Sci. U.S.A 1971, 68, 3195. 7. Grover, P. L., Sims, P. Biochem. Pharmac. 1973, 22, 661. 8. Rogan, E. G., Cavalieri, E. Biochem. Biophys. Res. Commun. 1974, 58, 1119. 9. Love, D. L. Naval Ordnance Laboratory technical report no. 74-197, Nov.
8, 1974. 10. Conney, A. H. Pharmac. Rev. 1967, 19, 317. 11. Busbee, D. L., Shaw, C. R., Cantrell, E. T. Science, 1972, 178, 315. 12. Whitlock, J. P. Jr., Cooper, H. L., Gelboin, H. L. ibid. 1972, 177, 618. 13. Levin, W., Conney, A. H., Alvares, A. P., Merkatz, I., Kappas, A. ibid. 1972, 176, 419. 14. Cantrell, E. T., Warr, G. A., Busbee, D. L., Martin, R. R. J. clin. Invest. 1973, 52, 1881. 15. Huberman, E., Sachs, L. Inc. J. Cancer, 1973, 11, 412. 16. Kellerman, G., Luyten-Kellerman, M., Shaw, C. R. Am. J. hum. Genet. 1973, 25, 327. 17. Kellerman, G., Shaw, C. R., Luyten-Kellerman, M. New Engl. J Med. 1973, 289, 934. 18. Löfroth, G., Natarajan, A. T. Läkartidn. 1975, 72, 593. 19. Korsgaard, R., and others. Unpublished. 20. Lancet, 1974, i, 910.
’Based upon 15 complete smoking histories. tBased upon 13 complete smoking histories; 1 non-smoker in this group. based upon 7 complete smoking histories; 1 non-smoker in this group. Classification in high, intermediate and low A.H.H. inducibility groups, and calculation of the expected numbers of patients in each group is according to Keller17 man et al.
for A.H.H. assessment. 38 patients left venous blood-samples which were assayed for A.H.H. inducibility at the department of tumour cytogenetics, Lund. Lymphocyte suspensions were prepared by a one-step centrifugal technique using ’Lymphoprep’, and the lymphocytes were then cultured in Parker 199 medium complemented with 15% fetal bovine serum, glutamine, antibiotics, and phytohmmagglutinin. After incubation at 37°C for 72 h 1-5 5 mmol/1 3-methylcholanthrene in acetone was added and allowed to take effect for an additional 24 h. The A.H.H. inducibility was then assayed by a modification of the method of Nebert and Gelboin.21 A high correlation between inducible A.H.H. activity in different cells from the same individuals has been demonstrated in previous studies.22 Our results are presented in the table. These preliminary data support the concept that A.H.H. plays an important role in the activation ofp.A.H. to ultimate carcinogens. The statisti-
cally highly significant (by chi-square test) over-representation of patients with high A.H.H. inducibility in this group of carcinomas might be of some value in identifying individuals at high risk. Mortality-rates of carcinomas of the respiratory tract are increasing, and screening of A.H.H. inducibilÜy23 24 in addition to sputum cytology, chest X-rays, and so on might improve survival figures, and even be of value in preventive medicine. This study was supported by grants from the Swedish National Association against Heart and Chest Diseases. R. K. is the recipient of a personal research grant (746-B74-02P) from the Swedish Cancer Socictv. Section of Preventive Medicine, Department of Internal Medicine and Department of Oto-rhino-laryngology, University of Lund, Malmö General Hospital, Malmö, Sweden; Research Department I and Department of Lung Medicine, University Hospital, Lund; and Department of Tumour
Cytogenetics, Wallenberg Laboratory, University of Lund
E. R. B. P.
TRELL
KORSGAARD HOOD KITZING GUNNELA NORDÉN B. G. SIMONSSON
COFFEE, MYOCARDIAL INFARCTION, AND SUDDEN DEATH
laboratory2 may shed light on the continuing controversy about the relation between coffee consumption and the incidence of myocardial infarction (M.l.) and subsequent death. Prospective studies26 27 have shown no corSIR,-Work in
our
21. Nebert, D. W., Gelbom, H. J. biol. Chem. 1968, 243, 6242. 22. Cantrell, E., Busbee, D., Warr, G., Martin, R. Life Sci. 1973, 13, 1649. 23. Chest, 1975, 67, 508. 24. Brooks, S. M. J. occup. Med. 1975, 17, 19. 25. Rotenberg, F. A., DeFeo, J. J. Fed Proc. 1976, 35, 384. 26. Dawber, T. R., Kannel, W. B., Gordon, T. New Engl. J. Med. 1974, 291, 871. 27. Klatsky, A. L., Friedman, G. D., Siegelaub, A. B. J. Am. med. Ass. 1973,
226, 540.
