Jan 28, 1980 - Marja Sorsa, institute of Occupational Health, Haartmaninkatu I , SF-00290 Helsinki 29, Finland. The adverse health effects of smoking have ...
Hereditas 92: 247-250 (1980)
Smoking and sister chromatid exchange KIRSTI HUSGAFVEL-PURSIAINEN, JORMA MAKLPAAKKANEN, HANNU NORPPA and MARJA SORSA Institute of Occupational Health, Helsinki, Finland
HUSGAFVEL-PURSIAINEN, K., MAKI-PAAKKANEN, J., NORPPA. H.and SORSA, M. 1980. Smoking and sister chromatid exchange. - Hereditas 92: 247-250. Lund, Sweden. ISSN 0018-0661. Received January 28, 1980
The present survey comprises analysis of sister chromatid exchange (SCE) frequencies in the peripheral blood lymphocytes of 100 subjects: 83 healthy men and women, both cigarette smokers and nonsmokers and 17 children. Both for men (30 smoking, 22 nonsmoking) and for women (13 smoking. 18 nonsmoking) the frequency of SCEs is significantly higher among smokers (group mean 9.6kS.E. 0.2) than among nonsmokers (group mean 8.1kS.E. 0.2). No difference is detected in the frequencies of metaphase chromosome aberrations analysed in the cultured lymphocytes of the same subjects. Young children (17 subjects, mean age 1.5 years) show a significantly lower frequency of SCEs (mean 5 . 1 ~ 0 . 6than ) adults. While the induction of SCEs is known to provide a sensitive indicator of mutagenlcarcinogcn exposure in experimental assays, it may also give important information of in vivo exposure to genotoxic agents. However, on the basis of the present data, which confirm previous results, the effects of individual smoking habits should carefully be taken into account in evaluations of the effects of exogenous agents on SCE frequencies.
Marja Sorsa, institute of Occupational Health, Haartmaninkatu I , SF-00290 Helsinki 29, Finland
The adverse health effects of smoking have been documented in thousands of studies during the last decades. According to harsh facts 30-50 % of cigarette smokers die of their smoking (DOLLand PETO1976). Cigarette smoke contains some 1200 different chemical entities (FALK1977). Several compounds, both in the gaseous and in the particulate phase of cigarette smoke, have been shown to be able to bind to cellular macromolecules either directly or after metabolic activation. Tobacco smoke condensate contains several known carcinogens, such as benzo(a)pyrene, benz(a) anthracene, crysene, p-naftylamine and "-nitrosonicotene (SCHMELTZ et al. 1974). Furthermore, phenols and amino acid pyrolysates are obviously formed during smoking through thermal degradation. Recent evidence suggests that phenols may have cocarcinogenic or cancer-promoting effects (SCHMELTZ et al. 1974) and that amino acid pyrolysates are active enhancers of mutagenicity in experimental systems (NAGAO et al. 1977). Consequently, even though the exact mechanism by This paper is dedicated to celebrate the 70th birthday of our much honoured teacher, Rofessor Esko Suomalainen, who has inspired our path in science at the Department of Genetics, University of Helsinki.
which cigarette smoking leads to lung cancer is still conjectural, the wide variety of biologically active compounds inhaled during smoking obviously provides inductive agents for both initiative and promotive steps of malignant growth. The objective for the present work has been to get information on the effect of smoking on sister chromatid exchange and chromosome aberrations in blood lymphocytes to settle previous discrepant et al. 1978; findings on the matter (cf. HOLLANDER LAMBERT et al. 1978; BALAKRISHNA MURTHY 1979) and to get necessary background data for our future work in genotoxicity of environmental agents in occupational settings.
Materials and methods Analysis of SCEs and chromosome aberrations
Whole blood lymphocyte cultures containing 2 ml Medium 199 (Gibco), I ml newborn calf serum (Microbiological Associates), 0.2 ml peripheral blood and 0.02 ml phytohaemagglutinin (Wellcome) were incubated at 37"C, 50 h for chromosome aberration analysis and 68 h in the presence of bromodeoxyuridine (BrdU, Calbiochem, 5.0
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K. HUSGAFVEL-PURSIAINEN ET AL.
,ug/ml) for analysis of sister chromatid exchanges. Colcemid (Gibco, 0.2pg/ml) was added into the cultures 2.5 h before harvest. Hypotonic treatment (8 min at 37°C) was performed with prewarmed 0.075 M KCI. The cells were fixed in methanol-acetic acid (3: 1) and the slides were made by the air-drying technique. Slides for SCE analysis were stained by the fluorescence-plus-Giemsa technique (PERRY and WOLFF1974) with slight modifications. The preparations were stained in Hoechst 33258 (1 pg/ml in Sgrensen buffer, pH 6.8) for I5 min, exposed to UV light (366 nm, at a distance of 3 cm) for 15 min, incubated in 2 x SSC solution at 60°C for 2 h and stained in 5 % Giemsa for 10 min (Merck, in Sgrensen buffer, pH 6.8). Preparations for chromosome aberration analysis were stained in % Giemsa (Merck9 in Sgrensen buffer? pH 7.0) for 5 min. *I1 'Iides were coded and loo metaphases per person were analysed for structural chromosome aberrations, and 30 harlequin-stained metaphases per person, for SCEs. The present material contains data Of made by four different persons in our laboratory. Smoking and nonsmoking sujects
SCE frequencies were analysed from 22 nonsmoking and 30 smoking men, average age 34 and 35 years, respectively, from 18 nonsmoking and 13 smoking women, mean ages 34 and 31 years, and from 17 children aged 2 to 60 months with an average age of 1 1/2 years. The subgroups among smokers and nonsmokers consist of male workers exposed to toluene in rotogravure plants, female workers exposed to Table I . Compiled data of smoking and sister chromatid exchanges (SCE) in adult subjects
Number of subjects
Number of SCE" cells analysed Group mean ?SE
22 18 40
758 538 12%
1.9
Women All Smokers Men Women All
30 13 43
878 450 1328
9s-r 9.9.. 9.bf"
Group
Nonsmokers Men
8.4 8.1
r0.2 r0.2
+0.2
styrene in reinforced plastic industry or to ethylene oxide in hospital laboratory, and of control persons occupationally unexposed to chemicals. All subjects were healthy according to personal interview and total blood counts taken. None of the smokers was a very heavy smoker (over 30 cigarettes per day); 19 men smoked 5 2 0 cigarettes per day, 7 men 1-19 cigarettes per day and 4 men were pipe smokers, while only 6 women smoked 5 2 0 cigarettes per day and 12 women 1-19 cigarettes per day. The chromosome aberration data included consist of the same occupational and control groups of men.
Results Compiled data of SCE frequencies among adult subjects, 40 nonsmokers and 43 smokers, are shown in Table 1. The group means (8,1-+0.2 for all nonsmokers and 9.6e0.2 for all smokers) differ statistically significantly.The SCE frequency in blood lymphocytes among the group of children studied (mean 5.1 kO.6) deviates significantly as compared to both of these values (Fig. 1) (SORSA et al. 1979). When the various exposure groups are compared (Table 2), the SCE frequencies are quite constant between the different groups of nonsmokers and smokers, respectively (the group of smoking, ethylene oxide exposed subjects is too small for a n y conclusions). However, the intragroup differences between smoking and nonsmoking subjects are statistically significantly deviant among unexposed male and female subjects (P
