268 Abstracts

expressed mucin gene in lung cancer, followed by MUC4; by contrast. the expression of MUC2 and MUC3 was almost undetectable in all cancer specimens. The intensity of expression of MUCI and MUC4 was always superior in cancer tissue than in the normal counterpart. As expected, the highest reactivity for MUCl and MUC4 expression was observed mainly in the adenocarcinoma histotype which is mucin secreting. These findings represent a contribution to the study of mucin gene pattern in lung cancer, and, in particular. indicate that MUC4, in association with the MUCl gene, seems to be strongly expressed in this neoplastic disease.

Detection of amplified genomic sequences in human small-cell lung car- cinoma cells by arbitrarily primed-PCR genomic fingerprinting Okazaki T, Takita J, Kohno T. Handa H, Yokota .I. Biobgy Diuision. Natl. Cuncer Ctr Research Institute, I-f Tsukiji .5-chome, Chuo-ku, Tokyo 104. Hum Genet 1996:98:253-S.

The arbitrarily primed-PCR (AP-PCR) genomic fingerprinting method was applied to evaluate its effectiveness in detecting and characterizing amplified DNA fragments in two small-cell lung car- cinoma (SCLC) cell lines, NCI-H69 and NCI-H82. Of the 2428 DNA fragments detected by AP-PCR using 62 arbitrary primers, 2 (0.08%) DNA fragments were amplified in NC&H69 and 6 (0.25%) DNA fragments were amplified in NCI-H82. Based on these results. we estimate the total size of the amplified genomic regions in these cell lines to be 3000 megabase pairs (Mt)) x 0.0008 = 2.4 Mb in NCI-H69 and 3000 Mb x 0.0025 = 7.5 Mb in NCI-H82. The 2 amplified frag- ments in NCI-H69 were mapped to chromosome 2, and all 6 amplified fragments in NCI-H82 were mapped 10 chromosome 8. This strongly suggests that restricted chromosomal regions are specifically amplified in these SCLC cell lines. Since the N-myc gene at 2~24 is amplified in NCI-H69 and the c-myc gene at 8q24 is amplified in NCI-H82, it is possible that these DNA fragments are co-amplified with N-myc or c-myc in these cell lines. However, since the 2 amplified fragments in NCI-H69 were not amplified in 42 other human cancer cell lines including 11 cell lines carrying amplified N-myc genes, it is also possible that there are amplified regions on chromosome 2 other than the N-myc locus at 2~24 in NCI-H69. In contrast, all 6 amplified fragments in NCI-H82 were amplified in several other human cancer cell lines carrying amplified c-myc genes. This result further indicates that these fragments were derived from an amplification unit that includes the c-myc gene. Our results show the ability of the AP-PCR method to analyze the fraction of the genome with amplification in human cancer CdS.

Transferrin dependence of Ga (NO,), inhibition of growth in human- derived small ceil lung cancer cells Weiner RE, Avis I, Neumann RD, Mulshine JL. Nuclror Medicine MC-2804. Connecticut University Health Center, 263 Farmington Ave. Fumingfun, CT 06030. J Cell Biochem 1996:62 Suppl. 24:276&87.

The effect of a combination of anti-transferrin receptor (TFR) anti- body, 42/6. and Ga(NO,), on cell growth was examined in small cell lung cancer (SCLC) cell lines: classic, NCI-H209, NCI-H345, NCIHS IO: and variant. NCI-H82 and NCI-N417. The role of Thit and

trans-ferrin (TF) in Ga(NO,), cellular uptake was also tested. Exoge- nous TF did not enhance the cytotoxicity of Ga. At > 3 pg/ml. Ga(NO,), inhibited growth in all cell lines in TF-supplemented or deficient media. At < 3 pg/ml. Ga stimulated growth for all cells but this effect was eliminated by TF or 42/6. Classic SCLC lines required 3- to 4.fold less exogenous gallium than variant lines to reduce cell number by 50%. The mean Ga uptake (“g/10’ cells) in H345 and H209 cell lines was 4- to 5-fold compared to H82 and N4 17 uptake (P < 0.001). 4216 reduced exogenous TF-stimulated growth. Antibody

plus Ga(NO,), caused a slight further cell number decline in all cell lines in TF-supplemented or deficient media. These results suggest that the addition of 42/6 antibody treatment would not increase the effec- tiveness of Ga(NO,), in patients. Both exogenous and endogenous Th and TFR play an important role in Ga uptake in these cells.

Monoclonal antibody aIR-3 inhibits non-small cell lung cancer growth in vitro and in viva Zia F, Jacobs S. Ku11 F Jr. Cuttitta F, Mulshine JL, Moody TW. NCI, BPRP, Bldg. C, 9610 Medical Ctr. Dr. Rocksiilr, MD 20850. J Cell Biochem 1996;62 Suppl. 24:269-75.

The ability of monoclonal antibody (mAb) aIR-3 to interact with non-small cell lung cancer (NSCLC) cells was investigated. MAb aIR-3 inhibited specific binding of [“‘I]IGF-1 and [‘*‘I]aIR-3 to a panel of 8 NSCLC cell lines with high affinity (IC,, = 200 and 50 “g/ml, respec- tively). [“‘I]-aIR-3 bound with high affinity (& = 40 “g/ml) to a single class of sites (B,,, = 8.000/tell) using NCI-H838 cells. [1251]aIR-3 was internalized when exposed to NCI-H838 or HI299 cells at 37°C but not 4°C. zIR-3 immunoprecipitated major 90 and 130 kD proteins. IGF-I stimulated and aIR-3 inhibited the clonal growth of NCI-H1299 cells. aIR-3 slowed the growth of NCI-H157 and H838 xenografts in nude mice. In a biodistribution study [“sI]rrIR-3 was preferentially localized to the tumor as opposed to other organs. These data suggest that IGF-I may be a regulatory agent in NSCLC.

GRP receptors are present in non small cell lung cancer cells Moody TW, Zia F, Venugopal R, Fagarasan M, Oje H, Hu V. NCI. EPRP Bldg. C, 9610 Medical Center Dr. Rockuille, MD 20850. J Cell Biochem 1996:62 Suppl. 24:247-56.

Previously, GRP receptors were characterized in small cell lung cancer cells and here non-small cell lung cancer (NSCLC) cells were investigated: [‘251-Tyr4]bombesin (BN) or [“sI]GRP bound with high affinity to NCI-H720 (lung carcinoid) and NCI-H1299 (large cell carcinoma) cells. Binding was specific, time dependent. and saturable. Specific [1X1-Tyr4]BN binding to NCI-H 1299 cells was inhibited with high affinity by GRP. BN, GRP’4-27, [D-Phe6]BNb-” methyl ester, moderate affinity by NMB, and low affinity by GRP’-‘6. BN (10 nM) transiently elevated cytosolic calcium in a dose dependent manner BN caused translocation of protein kinase C from the cytosol to the membrane and the translocation caused by BN was reversed by [D- Ph&‘]BN’-” methyl ester. BN stimulated arachidonic acid release and the increase caused by BN was reversed by [D-Phe6]BN6-13 methylester Using a clonogenic assay, BN stimulated the growth of NCI-H720 cells, and the number of colonies was reduced using [D-Phe6]BNb-‘3 methylester. These data suggest that GRP receptors that are present in lung carcinoid and NSCLC cells may regulate proliferation.

Novel metastasis model of human lung cancer in SCID mice depleted of NK cells Yano S, Nishioka V, Izumi K et al. Third Depr oflntrrnal Medicine, Tokushimo Uninitl. School of Medicine Kuramoto-cho 3, Tokushima 770. Int J Cancer 1996;67:211-7.

Metastasis is a critical problem in the treatment of human lung cancer. Thus. a suitable animal model of metastasis of human lung cancer is required for in viva biological and preclinical studies. In this study, we tried to establish a suitable model for this, using SCID mice. Neither human SCLC H69/VP cells (5 x 106) nor squamous-cell car- cinoma RERF-LC-AJ cells (1 x 106), injected through a tail vein, formed metastases in untreated SCID mice. Pretreatment of SCID mice with anti-asialo GM, serum resulted in only a few metastases of H69lVP cells, but pre-treatment with anti-mouse IL-2 receptor 13