110 Abstracts 1 Lung Cancer 16 (1996) 1OSlZ7

Zhou J, Mulshine JL, Unsworth EJ et al. Biomarkers/Prevenr;on Res. Branch. DCS, NCI, 9610 Medical Center DE. Rockville, MD 20850- 3300. J Biol Chem 1996;271:10760-6.

We have reported that a mouse monoclonal antibody, 703D4, de- tects lung cancer 2 years earlier tban routine chest x-ray or cytomor- phology, We purified the 703D4 antigen to elucidate its role in early lung cancer biology, using Western blot detection after SDS-polyacry- lamide gel electrophoresis. Purification steps included anion exchange chromatography, preparative isoelectric focusing, polymer-based C,,- like, and analytical C, reverse phase high performance liquid chroma- tography. After 25-50,000 fold purification, the principal immuno-stain- ing protein was >95% pure by Coomassie staining. The NH, terminus was blocked, so CNBr digestion was used to generate internal peptides. Three sequences, including one across a site of alternate exon splicing. all identified a single protein, heterogeneous nuclear ribonucieopro- tein-A2 (hnRNP-A2). A minor co-purifying immunoreactive protein resolved at the final C, high performance liquid chromatography step is the splice variant hnRNP-B 1. Northern analysis of RNA from pri- mary normal bronchial epithelial cells demonstrated a low level of hnRNP-A2/T31 expression, consistent with immunohistochemical stain- ing of clinical samples, and increased hnRNP-A2/T31 expression was found in lung cancer cells. hnRNF’- A2/Bl expression is under prolif- eration-dependent control in normal bronchial epithelial cell primary cultures, but not in SV40-transformed bronchial epithelial cells or tumor cell lines. With our clinicai data, this information suggests that hnRNP- A2031 is an early marker of lung epithelial transformation and car- cinogenesis.

Atypical adenomatous hyperplasia and bronchoalveolar lung carcinoma: Analysis by morphometry and the expressions of ~53 and carcinoembryonic antigen Kitamura H, Kameda Y. Nakamura N. DeportmentoJPathologv. School ojilfedicine. Yokohama City Universip, 3-9 Fukuura. Kanazawa-ku, Yokohama 236. Am .I Surg Path01 1996;20:553-62.

Atypical adenomatous hyperplasia (AAH) of the lung is a putative precursor of bronchoalveolar carcinoma (BAC). To define the steps in its development and to clarify at which stage critical cellular events occur. we studied 65 lesions of AAH, early BAC. and overt BAC by morphometric analysis and immunohistochemical evaluation of expres- sion of ~53 protein and carcinoembryonic antigen (CEA). Both the nuclear area and lesion size increased from AAH to early BAC and to overt BAC: the standardized variation of nuclear area was smallest in overt BAC. Discriminant analysis using these morphometric param- eters revealed high accuracy rates for the respective categories. Analy- sis of distribution of lung lesions in terms of nuclear area and lesion size yielded effective, potentially diagnostic cutoff values for distinc- tion between AAH and early BAC. Both ~53 and CEA expression tended to increase with the advance of atypia grade. In particular, high-level ~53 expression was strongly correlated with overt BAC. These find- mgs indicate that our classification of lung lesions is reproducible and thus useful for analyzing the development of BAC. Furthermore, some kinds of p53 gene abnormalities that are correlated with high-level ~53 expression likely play an important role in the progression of early to overt BAC.

Selective maternal-allele loss in human lung cancers of the ma- ternally expressed pS7(KIPf) gene at llp15.5 Kondo M, Matsuoka S, Uchida K. Laboratorv CUtraslructuw Research, ,4ich; Cancer CenterReslnshtute. Chrkusa-ku, Nagoya 464 Oncogene 1996;12:1365-8.

Genomic imprinting at 11~15 is suggested to play a role in ten pediatric tumors such as Wilms’ tumor, based on the findings of se tive maternal loss of this chromosomal region. Although the allele at llpl5 is also frequent in a number of cancers of adults includ lung, breast, and bladder cancers, possible involvement of geno imprinting in these tumors has not been investigated extensiv p57(KIP2), a newly described member of the p2 1 cyclindependenl nase (CDK) inhibitor family which is thought to negatively regu the cell cycle at the G, checkpoint, has been mapped to 11~15. In present study, we searched for somatic p57(KIP2) mutations in I cancer, but failed to find such alterations. Interestingly, however, found that the p57(KlP2) gene is imprinted with maternal expres: and that the maternal alleles had been selectively lost in 11 of 13 (8 lung cancer cases carrying 11~15 deletions, this being a signifil bias (P = 0.01). These data provide the first evidence that geno imprinting may play a role in the oncogenesis of not only rare pedi; tumors but also this common cancer of adults, suggesting that the printed p57(KiP2) CDK inhibitor gene is a potential target for m: nally biased 11~15 deletions.

P21 RNA and protein expression in non-small cell lung ca nomas: Evidence of p53-independent expression and asso’ tion with tumoral differentiation Marchetti A, Doglioni C, Barbareschi M. lnst Pathological Anat,

Hmtologv Unrversity of P&a, via Rome 57, 56100 Pisa. Oncog 1996;12:1319-24.

~21, the product ofthe WAFlICIPl/SDlllmdad gene, is an in1 tor of cyclindependent kinases. In cell cultures p2 1 is induced by 1 dependent and p53-independent pathways by DNA damage and inc tion of differentiation. We investigated p2 1 RNA and immuno-hl chemical expression in 43 non-small cell lung carcinomas and cc sponding normal lung samples previously investigated for ~53 WAFl gene status and ~53 protein expression. p21 RNA and prc expression in normal and neoplastic tissues were strictly associate = 0.0001). In normal tissue p21 RNA was expressed at low levels p2 I immunoreactivity was seen in scattered differentiated broncl alveolar and stromal cells. In the majority of neoplasms p21 prc and RNA were expressed at higher levels than in the corresponl normal tissues: p21 overexpression was seen in 27 (63%) and 28 (6 cases, respectively. p2 1 was expressed independently from ~53 g protein alterations. p21 overexpression was more frequent in well ferentiated tumors (P = 0.01 and P = 0.022 for RNA and proteir spectively), and p21 immunoreactivity was usually seen in foci of I pronounced differentiation. We conclude that p2 1 expression is rel to tumor differentiation, and that p53-independent p21 expression common feature of in vivo neoplasms.

Pathology

CD44v6 expression in primary bronchioloalveolar carcinc and conventional pulmonary adenocarcinoma Ohori NP. Coppola D, Landreneau RI. Monte/Tore-Pathotogv, Unr

siw of Pltrsburgh. Medrcal Center, 200 Lothrop &reef, Pittsbqh 15213-2582. Mod Path01 1996;9:507-12.

CD44 is a family of transmembrane glycoproteins involved in to-cell and cell-to-matrix interactions. Of the CD44 isoforms chz terized, the v6 variant has been shown to confer metastatic potenti animal models and its expression has been correlated with aggre!