272 Abstracts/Lung Cancer 12 (1995) 265-329

Deteetkm of intratmnoral DNA heterogeneity in primary lung cancer using a multiple sampling method Mizobuchi K, Fuji& Y, Hsshikurs H, Arimoto T, N&g&i Y, Goto T et al. Second Department of Medicine. Kyoto Prefectural Univ. o/Medicine, Kumigvoh, Kyuto. Jpn J Thorae Dis 1994;32:963-9.

The nuclear DNA conteots of 18 freshly resected speeimcns of primary lung cancer tissue were determined by flow cytometry with s multiple sampling method. A DNA sncuploidy pattern wss observed in all cases. In three cases, s diploid DNA pattern wss observed along with abnormal DNA stem lines. Twelve cescs (66%) had two or more abnormal DNA stem lines, and were classified ss DNA multiploidy. The frequencies of DNA ancuploidy and DNA multiploidy were higher than in previously reported studies. Intrstumoral DNA heterogeneity wss observed in IO esses (55%). Three of these were s mixture ofDNA diploidy and DNA encuploidy, and the other seven were B mixture of DNA aneuploidy and DNA multiploidy. If multiple sampling had not been performed in these cases, their ploidy patterns would hevc been misinterpreted. We conclude that multiple sampling is important in determining the precise DNA ploidy patterns of primary lung cancers.

Analysis of DNA ploidy with bmncboscopic brushing specimen as a diagnostic aid for lung cancer Kim YC, Lee SS, Chung IJ, Kang YH, Choi IS, Park KO et al. Deparlmenr of InternalMedicine, College ofMedicine, Chonnam University. Kwongiu. Tuberc Respir Dis 1994,41:354-62.

Objectives and m&/w& The presence of aneuploidy or high proliferative activity in cytologic specimens is considered BS complementary for the diagnosis of malignancy. To evsluste the diagnostic usefulness of DNA ploidy and cell cycle analysis in lung cancer, we compared the diagnostic yielding rates of DNA ploidy test by brushing specimms using flow cytometry with bronchoscopic forceps biopsy and bmshing cytology. Resulfs: Of the seventy-six cases, 55 csses proved to have malignant diseases (squamous cell cancer: 27, adenocarcinoma: 7,lsrge cell cancer: 1, undifferentistedz 4 and small cell csocer: 16). The incidence of sneuploidy in lung csncer patients wss 32.7% (1 B/55), ss opposed to no cases in benign disease. And the proportion of high proliferative activity (S+G2M>22%) in lung cancer patients wss 42.9% (15/35), but none in benign diseases. In fifty-six of 75 eases (74.7%). cytology of brushing speeimcns and DNA analysis (either aneuploidy or high proliferative activity vs. diploidy and low proliferative activity) were in concordencc. The sensitivity with only brushing cytology was 41.8% (23/55), but with the addition of DNA analysis, it wss iocreased to 56.4% (3l/55), without decreasing the speeiticity (100%). And there wss II CBS= whose clue for malignancy wss absent except ancuploidy, and he ws confirmed to have squamous cell esncer following open thoracotomy. Then were no differences in the frequeocy of ancuploidy or high proliferative activity betwax histologic subtypes of bronchogenic malignancy. Conc1u.sion.v: The diagnostic detection rate of lung caoeer wss improved with the addition of DNA ploidy and cell cycle analysis, and the presence of ancuploidy or high proliferative activity wss s relatively specific indicator of malignant disease. It would be useful to test DNA ploidy and cell cycle analysis with brushing speeimcn for the diagnosis of bronchogenic malignancy particularly in patients whose biopsy specimen could not be obtainable.

C~~dpYpm(einowrerp~oqgeaemutatimwitbp~ in resected non-small cell kmg cancer (NSCLC) patients Lee YH, Shin DH, Kim JH, Lim HY, Chung KY, Yang WI et al. Department of PulmonqMedicine, Ajou Unimsiry SchoolojMrdicine. Suwon. Tubere Respir Dis 1994;41:339-53.

Bu+nund: The ~53 gene codes for s DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the Gl to the S phase of the cell cycle. Mutations of the ~53 gene are common in B wide variety of human esncers, including lung csncer. In lung CBIICOR, point mutations of the ~53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the ~53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of ~53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus end cso be detected by immuno- histoehcmical staining. But protein overexpression has been reported in the absence of mutation. ~53 protein overexpression or gene mutation is reported

poor prognostic factor in bresst cancer, but in lung cancer, its prognostic significance is controversial. M&hod: We investigated the ~53 abnormslities by nucleotide sequencing, polymcrase chain reaction-single strand conformation polymorphism @‘CR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the ~53 protein wss studied immunohistoehemicslly in archival paraff~n-embedded tumor samples using the DO7 (Novocastra, U.K.) antibody. Overexpression of ~53 protein wss defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection ofp53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of ~53 gene. Result I) Of the 75 patients, 36% (27n5) showed ~53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative ~53 immunostaining (overall median suwivsl of26 months, disease free median survival of I3 months in both groups). 2) By PCR-SSCP, 27.6% (161 58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift (overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3)Nucleotide sequence wss analysed from 29 patients, and 34.5% (10129) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutstion (overall median survival of 22 vs 27 months, disesse free median survival of IO vs 20 months), but there wss no statistical significance 4) The sensitivity sod specificity of immunostain bssed on PCR-SSCP wss 67.0%. 74.0%, and that ofthe PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP wss 62.5%, and the rate bctwcen the PCR-SSCP and nuclcotidc sequencing wss 95.3%. Conclusion: In terms of detection of ~53 gene mutation, PCR-SSCP wss superior to immunosteining. p53 gene abnormalities either overexpression or mutation were not s significant prognostic factor in NSCU: patients resected witb curative intent. However, patients with the mutated ~53 gene showed the trends of early relapse.

Lack of tissue specificity of mucin markers in a lung cancer model: Bioehemicnl nppmach Bombardieri E, Sercgni E, Bogni A, Botti C. Maftioli L, Lombard0 C et al. Divkionc di Medic&m Nucleore. Isrihrro Naz. per lo Studio, la Cum dei Tumori. vi0 G. Venezian I. 20133. M&ma Int J Oncol 1994;5:1363-7.

Mucin-associated epitopes we recognized by monoclonsl antibodies in the immunometric assays used for the diagnosis and monitoring of eaneer. The recently developed new assays messwe mu&s ss tumor markers, assuming that each mucin is associated with s particular tumor site, i.e. CA 15.3 and MCA with breast eaocer, CA 125 with ovarian cancer, CA 19.9 and CA I95 with colon and pancreatic cancer. These associations are based on the frequency and the intensity of expression of the single markers for s certain organ. However, this theoretical orgsn specificity is not absolute, sinw the mucins are expressed also by tumors other than those mentioned above and they may also be present in inflammatory conditionsand in normal tissues. These observations were mntirmed by the present study, which used so experimental model consisting of s pool of 20 lung tissue samples (10 normal sod IO cancer). The tissue concentrations of the mucins MCA, CA 15.3, CA 125, CA 19.9, and of the glycoprotein CEA, were measured both in malignant tissue samples and in their normal counterparts. The marker levels were detected by immunometric assays in mucin fractions sepsrated from the tissue extract by chromatogmphic methods. The comparison of the chromatogrsphic profiles and the eveluation of the mucin levels in normal and malignant lung tissue specimens confirmed the absence of tissue specificity of these biochemical parameters. Recent developments in molecular biology and the diseovety of genes coding for several spomucins may open new perspectives towards the understanding of the mechanisms regulating mu& pathways.

Somatostath receptor expression ia kmg cancer O’Byme KJ, Halmos G, Pinski J, Groat K, Szepeshszi K, Schally AV et al. ICRF Clinical Oncology Unil, Churchill Hospital, Headington. Oxford OX3 7LJ. Eur J Cancer Part A Gen Top 1994;30:1682-7.

Experimeotsl evidence suggests that somatostatin anslogues may have B role to play in the mansgement of lung tomours. We evaluated membrane preparations of nine small cell lung caocer (SCLC) cell lines and of tomour samples from 3 patients with non-small cell lung career (NSCLC), I patient with so atypical carcinoid and another with s bronchial carcinoid for the presence of specific