Celotti L, Furlan D, Ferraro P, Levis A G
Department of Biology, Padova, Italy.
Mutagenesis. 1989 Mar;4(2):82-6. doi: 10.1093/mutage/4.2.82.
In a sample of healthy subjects (23-59 years old), comprising 49 non-smokers and 23 smokers, we studied the effects of smoking on DNA repair synthesis (UDS) in peripheral lymphocytes. DNA repair after UV-irradiation was measured by [3H]TdR incorporation into DNA (semi-conservative synthesis was inhibited by hydroxyurea). Compared to lymphocytes from non-smokers, lymphocytes from smokers showed significantly higher levels of UDS and higher incorporation of [3H]TdR in non-irradiated cells. We also determined the rate of DNA replication after 60 h in the presence of phytohaemoagglutinin (PHA) by measuring the incorporation of [3H]TdR during the last 20 h of culture. By irradiating the lymphocytes with UV light before setting up the cultures, we measured the inhibition of DNA replication due to damage induced by UV light. This inhibition, determined by the ratio of DNA replication in irradiated over non-irradiated lymphocytes, was less marked in smokers than in non-smokers. In lymphocytes from a portion of the donors (18 non-smokers and 19 smokers) we also determined the rate of the [3H]TdR incorporation in the absence of PHA and compared the distribution of the radioactivity between the two groups. In unstimulated lymphocytes, more radioactivity was incorporated by smokers compared to non-smokers in both conditions (UV-irradiated or mock-irradiated), while in stimulated lymphocytes the distribution of radioactivity was significantly different only after UV-irradiation.
在一个由49名不吸烟者和23名吸烟者组成的健康受试者样本(年龄在23至59岁之间)中,我们研究了吸烟对外周淋巴细胞中DNA修复合成(UDS)的影响。通过将[³H]TdR掺入DNA来测量紫外线照射后的DNA修复(羟基脲抑制半保留合成)。与不吸烟者的淋巴细胞相比,吸烟者的淋巴细胞在未照射细胞中显示出显著更高的UDS水平和更高的[³H]TdR掺入量。我们还通过测量培养最后20小时内[³H]TdR的掺入量,测定了在植物血凝素(PHA)存在下60小时后的DNA复制速率。通过在培养前用紫外线照射淋巴细胞,我们测量了紫外线诱导损伤对DNA复制的抑制作用。这种抑制作用由照射淋巴细胞与未照射淋巴细胞中DNA复制的比率确定,吸烟者中的抑制作用不如不吸烟者明显。在一部分供体(18名不吸烟者和19名吸烟者)的淋巴细胞中,我们还测定了在没有PHA的情况下[³H]TdR的掺入速率,并比较了两组之间的放射性分布。在未刺激的淋巴细胞中,在两种条件下(紫外线照射或假照射),吸烟者比不吸烟者掺入更多的放射性,而在刺激的淋巴细胞中,只有在紫外线照射后放射性分布才存在显著差异。