Summary We studied 100 patients with non-Hodgkin's lymphoma, 44 patients with Hodgkin's disease and 100 controls for the prevalence of the EcoRI restriction fragment polymorphism of the L-myc oncogene. No difference in the frequency of the three genotypes (LL, LS, SS) was found between the patient and control groups. However, the S allele was found to occur more frequently in the non-Hodgkin's lymphoma patients (X2 = 4.57, P = 0.032). These data confirm an earlier report and suggest that the presence of the S allele is associated with susceptibility to non-Hodgkin's lymphoma.Associations between restriction fragment length polymorphisms (RFLPs) of known oncogenes and a predisposition to develop cancer have been reported by a number of authors (Krontiris et al., 1985;Lidereau et al., 1985;Heighway et al., 1986). One of the most extensively studied oncogene RFLPs is an EcoRI RFLP of the L-myc oncogene. Digestion of DNA with EcoRI results in two alleles of 10 kb (L) and 6.6 kb (S). The S allele is reported to occur more frequently in male patients with bone and soft-tissue sarcomas (Kato et al., 1990), in oral cancer patients with poor to moderately differentiated tumours (Saranath et al., 1990) and in patients with leukaemia and lymphoma (Chenevix-Trench et al., 1989). Furthermore, an increased susceptibility to metastasis in lung (Kawashima et al., 1988), renal (Kakehi & Yoshida, 1989) and gastric cancer ) is associated with the presence of the S allele. However, a negative association between an increased susceptibility to metastases in Norwegian lung cancer patients and the S allele has been reported (Tefre et al., 1990). In this study we have investigated the frequency of the S allele in non-Hodgkin's lymphoma (NHL) patients and Hodgkin's disease (HD) patients compared with controls. al. (1988). Briefly, 5 ml of peripheral blood was lysed with 45 ml of cold lysis buffer (0.32 M sucrose, 10 mM Tris-HCl pH 7.5, 5 mM magnesium chloride and 1% Triton X-100) and then centrifuged for 10 min at 1,000 g. The pellet was resuspended in 5 ml of 4.0 M guanidine isothiocyanate, 25 mm sodium acetate and 0.84% P-mercaptoethanol and rocked gently for 20 min. The DNA was precipitated by the addition of an equal volume of isopropanol and resuspended in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0). A 3 ytg aliquot of DNA was digested with EcoRI, electrophoresed in a 0.8% agarose gel, transferred to Hybond N+ nylon membrane by alkaline transfer (Chomczynski & Qasba, 1984) and hybridised with a 32p_ labelled probe prepared by the random priming method of Feinberg and Vogelstein (1983). A 1.8 kb SmaI-EcoRI Lmyc fragment, pJB327 (Nau et al., 1985), excised from low gelling temperature agarose, was used as the probe. Washes were carried out at a final stringency of 0.3 x SSC and the autoradiographs exposed for 1-3 days on Kodak XAR-5 film at -80°C.
Materials and methods
Patients and controls
StatisticsThe frequency of the three genotypes and the two alleles in the patient and control groups were compared using the X2 test.
Results...