384 Abstracts / Lung Cancer 15 (1996) 381-399

The identitication of biomarkers to complement pathological stage for a more accurate prognosis and help clinicians decide on treatment is still an open problem for patients with lung cancer. Expression of P53 protein was detected by an immunohistochemical approach using the monocIona1 antibody PAbl801 on parafRnembedded sections of tumours obtained surgically from 102 stage II-IIIa patients with non- small-cell lung cancer (52 squamous cell carcinomas, 50 adenocarcinomas). [‘I-QThymidine labelling index, an indicator of the S-phase cell fraction, was evaluated on histological sections of [Wlthymidine-labelled tumour samples. DNA ploidy was defined by flow cytometric analysis on frozen tumour tissue. The biomarkers, histology and pathological stage were analysed in relation to relapse- free survival in univariate and multivariate analyses. Stage and interaction between [‘Hlthymidine labelling index and histology provided significant prognostic information for the overall series. [‘H]thymidine labelling index was an independent prognostic indicator of 3 year relapse-free survival in patients with adenocarcinoma. The results indicate the importance of cell proliferation to complement prognostic information provided by pathological stage in patients with stage II-IIIa adenocarcinomas.

Sex chromosome abnormalities in lung cancer patients Dave BJ. Hopwood VL, Spitz MR, Pathak S. Cellular Genetrcs Laboralory. M.D. Anderson Cancer Center, Box 181, 151.5 Holcombe Boulevard, Houston, Zf 77030. Cancer Genet Cytogenet 1996;87:24- 8.

Chromosomal analyses of lymphocytes from lung cancer patients and normal subjects revealed that the X and the Y chromosomes have both structural and numerical abnormalities in higher frequency in patients compared to the controls. These abnormalities included chromatid/isochromatid breaks. translocations, ring formation, and selective nondisjunctions, resulting in multisomies of either the X or Y chromosomes. Possible significance of these genetic abnormalities are discussed in relation to lung cancer patients.

Microsatellite instability and loss of heterozygosity at the hMLH1 locus on chromosome 3~21 occur in a subset of non- small cell lung carcinomas Wieland I, Ammermuller T, Bohm M. Totzeck B, Rajewsky MP. Cell Biology 1ns1. (Cancer Research), University of Essen Medical School, Mrchowstr 173, D-45122 Essen. Oncol Res 1996;8: l-5.

Microsatellite alterations observed in tumor specimens may reIlect genomic instability due to defective mismatch repair genes. To investigate whether this occurs in human nonsmah cell lung carcinomas we have analyzed microsatellite instability at 5 marker loci and loss of heteroygosity (LOH) at the hMLH1 locus on chromosome 3~21 using the polymerase chain reaction, Gf a total of 49 nonsmall cell lung carcinomas examined, 43% (13 of 30 informative cases) showed LOH at 3~2 I and 29% (I4 of 49) exhibited microsatellite instability at one or multiple loci. LGH of the mismatch repair genehMLH1 at 3~21 occurred in 82% (9 of I I informative cases) of the tumors with microsatellite instability. This suggests that defects in the mismatch repair gene hMLHl at 3~21 may be involved in microsatellite instability and tumorigenesis of a subset of nonsmall cell lung carcinomas. However. in those nonsmall cell lung carcinomas without microsatellite instability LOH at 3~21 probably involved another tumor suppressor gene(s) in this chromosomal region.

Characterization of delta opioid receptors in lung cancer using .a novel noqqtidic ligand Campa MJ, Schreiber G, Bepler G, Bishop MJ, McNutt RW, Chang K- Jet al. Duke VniversilyMedicol Cente,: Box 3808, Durham, NC 27710 Cancer Res 1996;56: 1695-701.

Cancer cells are often characterized by the presence of membrane receptors not normally associated with nontransformed cells from the same tissue type. Recent studies have demonstrated increased expression of high-a&thy binding sites for opioid receptor-selective ligands in lung cancer cell lines relative to normal lung tissue. We investigated the binding of a norqwptidic delta opioid receptor ligand in small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) cells with the aim of developing the Iigand as a novel lung cancer imaging agent. The @and, [‘I-B (+)-4[(a-R)a-((2&5R)-4- allylt,5-dimethyl- l-piperazinyl)-3-hydroxybenzyl)N,N-diethylbenzamide ([‘H](+)BW373U86). bound with high-affinity [K(d) (dissociation constant) = 0.066 f 0.012 t&B to membranes prepared from six different SCLC cell tines but not to those from seven NSCLC cell lines, including one mesothelioma. The number of binding sites varied from IO to 300 fmohmg membrane protein. Competition binding studies demonstrated displacement of [‘H](+)BW373U86 binding by the delta-selective antagonists naltribcn and 7benzylidenenaltrexone but not with the i- and kappa-selective antagonists D-Phe-Cys-Tyr-D-Trp-Gm-Thr-Pen- Thr-NH, and trans-(*)-3,4-dichloro-N-methyl-N-(2-(1- pyrrolidinyl) cyclohexyl]benzeneacetamide methanesulfonate. Mean apparent K(i)s for naltriben and 7-benzylidenenaltrexone in membranes from two SCLC cell lines were 0. I7 and 3.9 nM, respectively, but were >I0 lM for the i and kappa ligands. The nonselective antagonist naloxone displaced [‘HJ(+)BW373U86 binding with an apparent K(i) of -29 nM. On the basis of these data, we believe the lung cancer receptor to be similar, if not identical, to the human brain delta opioid receptor. The lack of high-affinity [‘H](+)BW373U86 binding in normal mouse lung mem- branes suggests a potential role for this ligand as a novel therapeutic or imaging agent.

Clonal evolution of lung tumors Chung GTY, Sundaresan V, Hasleton P, Rudd R, Taylor R, Rabbitts PH. MRCCORU. Medical Research Council Centre, Hills Rood. Cambridge CB2 2QH. Cancer Res 1996;56: 1609-14.

Lung tumors, particularly squamous cell carcinomas, are believed to develop through a series of morphological abnormalities, driven by underlying somatic genetic changes, One way of studying this process is to analyze candidate somatic genetic changes in samples of squamous metaplasia and bronchial dysplasia of varying degrees of severity as well as tumor from the same patient. This assumes a clonal relationship between these lesions. In this article, we provide evidence that adjacent, physically distinct bronchial abnormalities are clonally related. This has been achieved using a plaque assay technique to detect the same ~53 mutation, present throughout a tumor specimen, in a small proportion of cells in an adjacent squamous metaplasia. In addition, we have obtained two dysplasia samples from a tumor- free patient over a 9.month interval. The earlier sample had one pS3 mutation, whereas the later sample had two ~53 mutations on different alleles, Thus. the pattern of clonal evolution detected in the parallel samples mimics the pattern seen in longitudinal samples and supports the analysis of synchronously collected samples for the study of tumor progression.