12

042

HAVE CYTOKINETIC PARAMET ERS OBTAINED FROM IN VIVO LABELED LUNG CANCER BIOPSIES PROGNOSTIC SIGNIFICANCE? G. ten Velde, M. Tinnemans, M. Lenders, B. Schutte, A. Schols, F. Ramaekers. Dept. of Pulmonology and Dept. of Molecular Cell Biology and Genetics. University of Limburg, PO Box 5800, 6202 AZ Maastricht, The Netherlands.

Conflicting data exist concerning the prognostic relevance of proliferative characteristics of lung cancer, such as DNA-index, the percentage of cells in S-phase and the ex-vivo tritiated thymidine labeling index. To overcome some of the drawbacks of these earlier studies, we examined biopsies obtained bronchoscopically from 59 patients with lung cancer, which were labeled in vivo with the DNA- precursor Bromodeoxyuridine (BrdU). Nuclear suspensions were stained with propidium iodide for DNA and anti-BrdU monoclonal antibody for DNA synthesis activity and analysed by bivariate flow cytometry.

Of these cases 48 were NSCLC and 11 SCLC. In 6 of these 59 patients the proliferative parameters could not be determined because of various reasons. Mean BrdU labeling index for SCLC was .5,3%, for NSCLC 4,0%, S phase transit time (Ts) 7,7 vs 7,4 hours and potential doubling time (Tpot) 167 vs 233 hours. A survival advantage has been seen in patients with lower LI, longer Ts and longer Tpot, although the differences are not significant.

In conclusion, in viva labeling of bronchoscopic specimens with BrdU produces adequate material for cytokinetic analysis of lung cancer, which can possibly be of prognostic significance.

044

POLYMORPHIC SITES WITHIN APC AND MCC LOCI REVEAL INFREQUENT LOSS OF HETEROZYGOSITY IN PRIMARY HUMAN NON-SMALL CELL LUNG CANCER. D. D’Amico, M. Spatafora, R. Bontade, E. Pace, D. Vitolo*, G. Bonsignore. Istituto di Fisiopatologia Respiratoria CNR, Palermo and *II Cattedra di Anatomia Patologica, Universiti di Roma “La Sapienza” - Italy.

Chromosome 5q21 deletion, as demonstrated by loss of hetemzygosity (LOH) for APC and MCC genes, has been recently described in 81% of small cell lung cancer (SCLC) cell lines and primary tumors (Cancer Res. 52, 1996, 1992).

The present study was aimed at evaluating the frequency of LOH for the APC and MCC genes in non-small cell lung cancer (NSCLC). Genomic DNA was extracted from archival partim-embedded tumor and normal autologous lung surgical samples (n=34) and used as template in polymerase chain reactions @‘CR) to amplify APC exon 11 and MCC exons 10, 12, and 15. The LOH was detected by diagnostic digestion of a Rsal polymorphic site located on APC exon 11, by PCR/SCCP analysis of polymorphic sites located within MCC exons 12 and 15 (Cancer Res 52,1996,1992), and by electrophoresis of PCR products of MCC exon 10 (Proc. Natl. Acad. Sci. USA 89,3385, 1992). 16/34 (47%) of the cases were informative at APC gene exon 11 Rsd restriction site and 2/16 (12.5%) of the informative cases showed LOH. 21/34 (61.7%) of the cases were informative for one or more MCC polymorphic sites and l/21 (4.8%) showed LOH. Collectively, the LOH for one of the investigated genes was found in 3/26 (11.5%) informative cases and in no case the LOH of both genes was detected in the same tumor sample.

We conclude that, unlike SCLC, the LOH of the putative tumor suppressor genes AK and MCC is a rare event in N-SCLC. These data support the concept of a genetic heterogeneity between NSCLC and SCLC.

043

THE STUDY OF HISTOPATHOLOGICAL FINDINGS AND NUCLEAR DNA PLOIDY PATTERN OF ADVANCED NON-SMALL CELL LUNG CARCINOMA (NSCLC) LOCATED PERIPHERALLY LESS THAN ZOMM IN DIAMETER. I. Nagahiro, M. Aoe, K. Okabe, S. Moriyama, A. Ando, N. Shimizu. Okayama University Medical School, Okayama, Japan.

The purpose of this study was to determine whether nuclear DNA ploidy pattern of NSCLC will affect the biological tumor attitude and patient survival rate.

From January 1982 through December 1991, there were 586 cases of NSCLC located peripherally and underwent surgical resection. In 130 cases of them, the tumor diameters were less than 20 mm ( 22.1% ). Among them. 19 cases ( 3.2% ) were diagnosed as advanced ( more than stage II ) pathologically because of lymph node metastasis, pleural dissemination, and intralobular metastasis. The histopathological findings, pathological stages, N-, D-, PM-factors, tumor differentiations, vascular invasions, tumor collagenizations, and nuclear DNA ploidy patterns were reviewed in these 19 cases. The mean age was 56.2 k 10.2 years old and there were 8 males and 11 females, 18 adenocarcinomas and 1 squamous cell carcinoma, and 2 stage II and 13 stage Ill and 4 stage IV cases. Ten cases showed diploid pattern, seven aneuploid, and others unclassifiable in their nuclear DNA ploidy patterns.

There were no correlations among each factors except between the nuclear DNA pattern and the vascular invasion (p=O.O13). In the aneuploid cases, the vascular invasion was observed more frequently than the diploid cases. Additionally, the survival rates were compared in the each factors. The d-years survival rate of diploid cases was 90.0% and aneuploid 21.4% respectively. There were no significant differences of the 5-years survival rates in the other factors.

These results indicate that the nuclear DNA ploidy pattern is a new factor that will be able to predict post-operative NSCLC patient’s survival.

045

LYMPHOCYTE SUBSETS AND NATURAL KILLER CELL PERCENTAGE IN BLOOD AND BRONCHOALVEOLAR

LAVAGE IN NON SMALL CELL LUNG CANCER (NSCLC)

??A. lXb1Ei Y, M. ,ilW SIII<lIA IA, 0.s. SAIAM.4, ‘A. KHALEI), and X’EIAER~A IR Y

* ‘I horaclc hlcd~cme and Clinical Pathology Departmcntr Mansoura University, Mansoura - Qypt

Lymphncytes have a central role in natural immunity against malignancy A prospeclive study was designed to assess the relation belwcen lymphocyle responses in NSCLC Total lymphocyte numbers and subsets (CD3, C’D4. CD4/CDI1 ratio and nntursl killer cell NK CD16 monoclonnl nntihodies) were measured in peripheral blood and bronchoalveolar lavage (BAL) 01’ IO crises squamous cell carcinoma (SQCC), 7 cases adenocarcinoma (ADC) and I Z healthy volunteers. The results in $Qcc peripheral blood total lymphocyte count, CD3%, CD4?/,, CD4/CD8 were significantly high while CD8?/. and CDl6% were significantly low in comparison to control Lavage cells were significantly higher except NK CDlb% which was significantly lower than control In ADC the rise was significant in peripheral blood total lymphocytes ~~o~CD:i’z&. Chile CD4 was lower hu: CTW? C4iCD8 and NK CD16% showed no dillbrence compared to control .-ihhough UAI. cells showed significant rise in total lymphocyte count. all subset% showed no difi‘erenco from that of control. There was no signiiicant WI I&:ion between simi!ar cell% in peripheral blood and in lavage

In conclusion lymphocyte count and its subsets showed variable cixnges in each histologic type of NSCLC. the changes include peripheral Ihlood cells nnd local lung cells with lack ofcorrelation between peripheral blood and nhl, WIIS