390 Abstracts/Lung Cancer 14 (19%) 377-408

embedded tumor samples. K-ras mutations were examined in 192 stage I to IV non-small-cell lung cancer patients. Results: K-ras mutations were detected in 51 of 192 of the cases studied (27%). All K-ras mutations detected by PCR/ AS0 hybridization were also identified by SSCP. In stage 1 disease, the median survival time was 46 months in those patients whose tumors had no K-ras mutations and 21 months in those with aspartic acid and serine mutations at K-ras codon 12; in patients with stage IIIA disease, median survival time was 16 months in the K-ras negative group and 7 months in the aspartic acid and serine mutation group. No significant differences were observed for the remaining amino acid substitutions of K-ras. nor were they observed at all in more advanced disease. Conclusionr: K-ras gene status has strong prognostic value in patients with stage IIIA non-small-cell lung cancer. The survival curve for patients with stage I and K-ras codon 12 aspartic or serine mutations isclose to that of patients with stage IIIA without K- ras mutations. However, a non-small-cell lung cancer K-ras genotypic classification should be validated in larger studies.

~53 nuclear immunostaining and gene mutations in non- small-cell lung cancer and their effects on patient survival Mitsudomi T, Oyama T, Nishida K, Ogami A, Osaki T, Nakanishi R et al. Department Surgery II, Faculty of Medicine, Kyushu University, 3-1-I Maidashi, Higashi-ku, Fukuoka 812. Ann Oncol 1995;6:Suppl 3:9-13.

Background: p53 gene mutations are known to occur in about half of all non-small-cell lung cancer (NSCLC) cases. Mutations of the p.53 gene usually but not always lead to an increased half life of the p53 protein, and result in a nuclear accumulation of protein which can be detected by imnumohistochemistry (IHC). Controversy still exists as to whether the presence ofan aberration of the ~53 gene or protein is a poor prognostic indicator in patients with NSCLC. Patients and methods: DNA samples and paraffin blocks were obtained from 129 patients of 143 consecutive patients who underwent a pulmonary resection during a 22-month period from July 1991 to April 1993. Mutations of the ~53 gene occurring at exons 5-8 were detected by a polymerase chain reaction(PCR)/singlestrandconformation polymorphism(SSCP) assay, while the nuclear accumulation of the p53protein was detected by immunohistochemistry. Results: Of the patients studied, 35 % had muta- tions and 54 % showed overexpression, when we defined a positive case as being one in which more than 10% of the tumor cell nuclei were stained. There was a 59.5% concordance between the ~53 gene muta- tions and ~53 imrnunopositivity. p53 immunopositivity in adeno- carcinoma and any ~53 abnormality (i.e. ~53 immunopositivity and/or mutation) inadenocarcinomawereapc~~prognosticindicator. However, Cox’s proportional hazards model indicated that the stage was the only significant prognostic factor. Conclusions: ~53 immunopositivity and mutations of the ~53 gene are frequently seen in NSCLC. They are considered to be mutually related but may sometimes represent a different aspect of p53 abnormality. ~53 alteration may be a poor prognostic indicator only in a subset of patients with NSCLC, especially for adenocarcinoma.

Molecular genetic tumor markers in the early diagnosis and screening of non-small-cell lung cancer Jacobson DR, Fishman CL, Mills NE. Research Service 151, New York V.A. Medical Center, 423 East 23 St, New York, NYloOlO. Ann Oncol 1995;6:Suppl 3:3-B.

Background: Little progress has been made in decreasing lung cancer mortality by applying conventional methods to early diagnosis and screening. Recent advances in molecular oncology, however, have provided tools which may be of use in this area. Many genes involved in controlling. cell growth and differentiation are abnortnal in lung cancer cells. Such genes include K-ras, ~53, rb, myc, her2/neu, and

probably one or more tumor suppressor genes on chromosome 3p. The involvement of these genes in lung cancer is reviewed. The K-ras oncogene contains a mutation in codon 12 in many cases of non-small- cell lung cancer, particularly adenocarcinoma, and is thus a potentially useful lung cancer tumor marker. Design: We have developed a highly sensitive, simpleassay forms mutations, andapplied it to bronchoalveolar lavage fluid obtained from patients undergoing evaluation for suspected lung cancer. Results: In many cases, the ras assay was more sensitive than routine cytology and histopathology, demonstrating that this is a potentially clinically useful assay. Conclusion: Molecular genetic tumor markers, including mutations in ras and other genes, and/or irnrnunohistochemical tumor markers, may provide tools which can be applied to bronchoalveolar lavage fluid or sputum, for use in diagnostic tests and in screening programs. The use of such markers may lead to decreased lung cancer mortality.

Analytical performances of Cyfra 21 and CEA assays with an I?S 300 Boehringer. Interest for the diagnosis and the follow- up patients suffering from NSC bronchial cancer GlikmanasG, Grivaux F, Lacroix I, Blanchon F, Thuillier F. Laboratoire de Biochimie. Centre Hospitalier, BP 218, 77108 Meaus Cedex. Immune-Anal Biol Spew 1995; 10:300-6.

Cyfra 21, detected in non-small cell lung cancer (NSCLC) patient’s sera, is a fragment of cytokeratin 19. An immunoradiometric assay (Irma) or an immunoenzymometric assay (Iema) was used with two specific, exclusive antibodies. We evaluated the Boehringer Iema with an ES 300 automatic system. Analytical study showed for intra-assay precision a variation coefficient (VC) < 5 46, for inter-assay precision a VC < 1096, and a detection limit < 0.6 rig/ml. The comparison with method (Irma CIS bio) was satisfactory. Also weshowed that conserving by freezing POD conjugate and biotinylated antibodies did not spoil antibody activity. Forty-six patients were tested, 41 with lung cancer and live with non-malignant pulmonary disease. For NSCLC, the diagnostic sensitivity of Cyfra 21 was 41.2% and for squmrous cell carcinoma 50%. Associated with ACE assay results, the sensitivity increased to 58.8%.

Selective suppression of cytokine secretion in patients with small-lung cancer Fischer JR, Schindel M, Stein LN, Lahm LH, Gallati H, Krammer PH et al. lhoras-Klinik, LVA Baden, Department of Medical Oncology. AmaiienstrasseS, 69126Heidelberg-Rohrbach. Ann Oncol1995;6:921- 6.

Background: Whether or not cytokine secretion is impaired in patients with small-cell lung cancer (SCLC), is unknown. We therefore investigated whether cytokine secretion by immunocompetent cells may be suppressed in patients with SCLC. Patients mzd methods: We determined cytokine secretion by lymphocytes and monocytes in whole blood cell cultures from 58 patients with SCLC, 95 patients with non- small-cell lung cancer (NSCLC), 10 patients with nonmalignant lung disease and from 44 nornral healthy individuals by using an enzyme- linked immunosorbentassay (ELISA) specific for thedifferent cytokines measured. Results: Compared to normal controls, inununocompetent cells from patients with SCLC secreted significantly lower amounts of IL-2, IFN (x, and IFN gamma upon mitogen stimulation. TNF (x- secretion was significantly reduced in SCLC extensive disease but not in SCLC limited disease. In contrast, secretion of IL-1 a. and IL-1 D was not reduced. In patients with NSCLC, secretion of IL-2 and IFN cx was signiticantlyreduced. ReductionofIFNgammasecretionwassignificant in metastasized NSCLC and marginally significant in localized NSCLC. Secretion of TNF a, IL-I a and IL-I R was not impaired. In addition, cytokina secretion in SCLC patients substantially improved upon