272 Abstracts/Lung Cancer 10 (1993) 266-286

al. Nat. Ins?. Occupational Health, Dcpamnent of Toxicology, PO Box 8149Dq. W33 Oslo. Environ Health Perspect 1992;98:187-9.

We~veexPminedrsstrictioaFrogmsDtlmgthpolymorphismsofche H-rss-I gene ia germ-lie DNA from 214 lung cancer patients sad 309 unaffected controls. When DNA ssmples were digested with MspI/ HpaII, Southern blot anslysis revealed at least 22 different alleles, grouped according to their frequencies as commoa. intermediate, and rare. The frequmcy of rare alleles ia hmg caacer patients (161428) is significantly different @ = 0.002) tium thst ia the control group (5/ 618). Individuals with rsre sllelea were found to bent 4.7-fold greater risk of lung cancer tbaa tbw with a0 rare sllele.5.

Activi~enhPManentofnllngesncer-~atedhumanmonoelwal antibody HB4CS by N-dqlyeosylalion Kate M, Mcchizuki K, Ha&nuns S, Tscbibsaa H, Sbirahsts S, Mursksmi H. Moriaaga 1n.a Biological Sclenn. 2-l-I Shimoweyoshi, Tswumi-ku, Yokohama23Ohum. AatibodiesHybridomss 1993;4:9-14

Ithnsbeen~ormthsttbelunguncer-associa~humnnmoa~loool antibodyHB4CScompriseshvolnmbdolightchPinsof30~ad32LDarrd that the 30 kD species is exclusively responsible for the aatibody activity. This study demoastrstes that the hvo light chsiis were both N- glycosylsted with glycosyl residues of different sizes, oae of which was sensitive to neuraminidsse sad the other insensitive. Our unpublished data of DNA sequence for the light cbsin of this monoclonal antibody indicated that the light chain cootaias only one possible site for N- glycosylation, which located ia the CDR-I. N-Deglycosylntion ofthis mcnoclonal antibody under the dtihuiag condition resulted in the co~leteconversionofthehvolightchsinsintooaeideoticPlpolypeptide chain of 26 kD. Activity of this monoclonsl sntibcdy wss found to be significantly enhanced by N-deglycosylstion. All the facts described above consistently indicate thst the activity of this monoclonal antibody is interfered with by the attachment of bulky glycosyl residues at the antigen binding site on tbe light &sin. The Ndeglywsylated antibody ws stable under the stwsge conditions employed, suggesting that the glycosyl residues attached to the light chain do not play any importzmt biological role except for interference with the antibody activity of binding sntigea.

PCR-based CYP2D6 genotyping for Finnish lung cancer patients Hirvonen A, Husgafvel-PursisinenK, AattilaS, Ksrjslsinen A, Pelkonen 0, Vainio H. Dept. of Industrial Hygiene/Taricol., Institute of Occuparional Healrh, Topeliukrenkaru 41 a A, SF-LX72SO HeLsinki. Phamracogenetics 1993;3: 19-27.

Polymorphism of the gene encoding for debrisoquine hydroxylase. i.e. CYP2D6, was determined genotypically for 122 healthy controls and 106 lung caacer patients using Xbs I restriction fragment length polymorphism (RFLP) snslysis, together with P combination of hvo recently published polymersse chain reaction (PCR) based approaches. Three different mutated alleles of the CYP2D6 gene were detected: CYP2D6B comprised 11.1% sad 10.4% of the total alleles in the controls and in the lung wncer patients, CYP2WA had frequencies of 5.7% and 2.8%, sad CYp2D6D had frequencies of 3.3% sad 2.4%, respectively. Only 17 of the 24 44 kb Xba 1 alleles (71%) were continned ss defective alleles carrying the mutation in CYPZD 68 loci, whereas sll four 15 + 9 Lb Xbn F alleles contsined the CYP2D6B mutation. Out of the 122 healthy controls, seven subjects (5.7%) were detected ss poor metabolizers (PM@ of debrisoquine by the presence of two defective alleles, whereas only one PM genotype wss found in the lung cancer pstient group (0.9%). The reliability of this analysis was confirmed in s subgroup of the control subjects phenotyped by debrisoquine, where s perfect correlation between CYF2D6 phenotype and genotypewas obtained. We observed no significant differencein the allelic frequencws between lung cancer patients with s history of heavy smoking and those who smoked less. However, statistical analysis showed s significant difference (p = 0.05) in distribution of the PM- associated genotypes between lung cancer patients (11106) and healthy controls (7/122). This data thus supports the hypothesis that there is an increased risk of lung cancer for individuals who sre extensive metsbolizers of debriscquine.

Expmssion of functional PDGF A receptors in a human large-cell ltmg-carcinoma cell line ForsbergK,Berghl,Westerms rkB.Depar?mer#ofPathology, Uniwrsity of Vppsala. 7518.5 Vppsala. Int 1 Csncer 1993;53:55660.

In this study we have investigated spsnel of lung-cancer cell lines, both of smsll-cell carcinoma and non-smallsell type, for the expression of receptors for plnteletderived growth factor. Although we found mRNA expression for the&typerweptoron ooesmsll-cell sndonenoa- small-eelllinepndbtypereceptormRNAexpressionononesmall_cell- cancer cell line, only the 8 receptors of the non-small-cell line (H-157) proved to be limctioasl. Thus, the cell line H-157 displayed specific binding of l”I-PDGF-BB in addition to mRNA expression of the 6-kb transcript for the PDGF R type receptor. Further evidence for the presence of functional PDGF 6 receptors in H-157 cells wss obtsined from sn in vitro kinsse assay, which demonstrated s ligsnd-induced receptor autophosphorylation ss well ss the phosphorylation of s number of potential substrates associated with the activated receptor.

dlk, A putatiremamnalii bomeoticgene differentially expressed in small cell lung carcinoma and neumeadacrine tumor cell line Labords J. Saasville EA. Hoffmsa T, Not&o V. CenterforBiologics and Research, FDA, Bldg. 29. 8800 Rockville Pike, Bethesda, MD 20892. J Biol Chem 1993;268:3817-20

GastrinrelePsingpeptideismitogenicformouseSwiss3’13fibroblasts sad certain humsa small cell lung carcinoms (SCLC) cells but not for mouse Bslb/c 3T3 fibmblasts. To identify new molecules sssocisted with the gsstrin relepsiag peptide-responsive phenotype, clones is&&d from P differurtinl cDNA library between Swis nod Balb/c 3T3 fibmblssts were used to screen for their expression in human SCLC cell lines. Using this spproach, we have isolated sad charscterized human and mouse cDNA clones encoding s novel protein. This protein is s putative trsnsmembnme protein belonging to the epidermal growth factor-like superfamily. In vitro tnmscription sad traaslstion studies detect s 42-kDn pmtein, in agreement with the size predicted from the translated cDNA sequence. This protein (termed D&s-like or dlk) is highly homologous to invertebrate homeotic proteins, includiig Delta, sad Notch, the products of neurogeaic loci involved in normal neural differentiation in Drosopbils. dlk is expressed in tumors with neuroendoxiae features, such BS neumblastoms, phmchromocytoms, and P subset of SCLC cell lines. However, its expression in normal tissues is restricted to the sdrensl gland and plscents. These data suggest that dlk may be iavolved in neumendc-xiae differentiation sad, because ofitscellulsrlocatioasnd restrictedexpressionin norms1 tissues, it msy be P potential tberspeutic target ia neumendoctie tumors, psrticulsrly SCLC.

Immunohistocbemical study of glutatbione-related enzymes and proliferativeantigeia lungcaneer: Relationtocisplatinsitivity Ogawa J-I, Iwszaki M, lnoue H. Koide S, Shohtsu A. First Depanment of Surgery. Tokai Vniwrsiry School of Medicine, Bohseidai, Isehara. Kat,agaw ZSPII. Cancer 1993;71:2204-9.

Background. With resected hmmr tissue from 84 patients with lung cancer, the expression of gluts&me peroxidase (GPX), glutsthioae reductase(GR),proliferatingcellnuclear~ntigen(PCNA),nndepidermal growth factor receptor (EGFR) wss examined in relation to cisdismminedichlomplatiaum(CDDP)sensitivity. Methods. TbeCDDP sensitivitywassssessed fmmsnincreaseofcells in the S-phaseor GZM- phase by s DNA histogram after the tumor cells were incubated in s CDDP solution. The expression of GPX, GR, PCNA, and EGFR for each tumor was studied with 811 indirect immuaoperoxidsse technique on psrsffiiembedded tissues. Results. The percentsge of pstients sensitive to CDDP according to the histologic type was 86% for smsll cell carcinomas, 40% for lsrgewll csrcinomss, 31% for squamous cell carcinomas, and 6% for sdenocarcinomss. There wss PO inverse relationship between CDDP sensitivity sad the frequency of GPX snd GR expression, sad both were significantly different smong histologic types. The GPX sad GR expression wss significantly lower in the CDDP-sensitivegroupthsnintheresistantgroup(P < 0.01). However. the expression of PCNA and EGFR was significantly lower in the sensitivegroupinnon-smaUceUcarc~omPsandsquPmousceUuuciaomas (P < 0.05). Conclusions. From tbe above findings, so