Summary To define the region on the short arm of chromosome 1 that is thought to include one or more tumour-suppressor genes for gastric cancers, we carried out loss of heterozygosity (LOH) studies in 26 gastric adenocarcinomas, using three restriction fragment length polymorphism (RFLP) markers and nine microsatellite markers. All tumours were informative with at least one locus; three revealed replication errors (RERs) at multiple microsatellite loci, and interstitial or telomeric allelic deletions were observed in 12 cases. Deletion mapping of these tumours defined a commonly deleted region between two loci, DlS201 and DlS197, that are 13 cM apart. As two loci within the commonly deleted region, D1S57 (pYNZ2) and D1S62 (pTHI54), were mapped respectively to lp35 and lp34.3 by fluorescence in situ hybridisation, we conclude that a locus likely to contain a tumour-suppressor gene for gastric cancer is located within a 13 cM region encompassing these two chromosomal bands. (Deng et al., 1987;Kihana et al., 1991) and p53 (Tamura et al., 1991;Strickler et al., 1994) genes. Amplification of the erbB-2 gene and mutations of the APC and ras genes are found frequently in well-differentiated adenocarcinomas but not in poorly differentiated adenocarcinomas (Yokota et al., 1988;Kihana et al., 1991;. In contrast, amplification of the K-sam gene and replication errors (RERs) at microsatellite loci have been detected preferentially in poorly differentiated adenocarcinomas (Hattori et al., 1990;Han et al., 1993). These observations imply that the genetic pathways involved in development of these two histopathologically distinguished forms of gastric adenocarcinoma are likely to be different.Recent results of LOH studies have suggested that loci containing tumour-suppressor genes associated with gastric carcinogenesis exist on chromosomal arms lp, lq, 5q, 7q, 12q 17p and 18q (Sano et al., 1991;Uchino et al., 1992; Kuniyasu et al., 1995). As part of a strategy to identify these putative tumour-suppressor genes we began by attempting to define the region on chromosome lp that is commonly deleted in gastric cancers. Here we report results of LOH studies in 26 well-differentiated adenocarcinomas of the stomach.
Materials and methodsPreparation of samples and DNA A total of 26 paired samples of tumours and corresponding normal tissues removed from Japanese patients with welldifferentiated adenocarcinomas of the stomach were obtained at the Cancer Institute Hospital, Tokyo. In 18 cases the tissue samples were fixed in formalin, embedded in paraffin and attached individually to glass slides. Genomic DNA was extracted according to methods described elsewhere (Goelz et al., 1985;Yanagisawa et al., 1991). The remaining eight samples were frozen in liquid nitrogen after surgical resection and stored at -80°C until isolation of DNA. Genomic DNA was extracted from the frozen tissues according to methods described elsewhere (Sato et al., 1990).RFLP markers for LOH analysis The three RFLP markers used in this study, D1S77 (pMCT58), DIS57 (...