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

cytokeratins 7 and 13. Among keratinixing squamous cell carcinomas, A rare case of primary choriocarcinoma of the lung in a male is all thymic carcinomas reacted with antibody specific for cytokeratin 7 described with immunohistochemistry forhuman chorionic gonadotropin (9/9, lOO%), whereas no staining reaction was seen in lung carcinomas (hCG), epidermal growth factor (EGF) and EGF-receptor. The extra- (O/S, 0%) (p < 0.01). This finding can be used as a diagnostic aid in gonadal trophoblastic origin of this pulmonary carcinoma was defmitely primary thymic keratinixing squamous cell carcinomas to expedite confirmed by an autopsy examination, and hCG-production and hCG- treatment and prognosis. Cytokeratin 7 and cytokeratin 13 monoclonal positive staining of the tumor cells. Furthermore, the tumor cells clearly antibodiesreactedwithalmostallcasesofthymiccarcinoma.Applications expressed EGF and its receptor which play an important role in the of monoclonal antibodies specific for certain cytokeratins, especially 7 proliferation and differentiation of normal and neoplastic trophoblasts and 13, may be helpful in the diagnosis of other subtypes of thymic of the UkNS. Our present case suggests that EGF may act in an autocrine carcinomas. manner in the tumor cells of primary pulmonary choriocarcinoma.

Standard and variant CD44 isoforms arecommonly expressed in lung cancer of the non-small cell type but not of the small cell type Arim A, Mate JL, Isamat M, Lopez D, Von Uexkull-Guldeband C, Rose11 R et al. Anatomic Pathology, Hasp Univ. Germans Trias i Pujol. 08916 Bodalona, Barcelona. J Path01 1995;117:363-8.

Cluster of differentiation 44 (CD44) encompasses a polymorphic family of cell membrane glycoproteins involved in the mechanism of tumour invasion and metastasis. Since non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) display very different rates of progression, a significant discrepancy in their CD44 expression profiles is to be expected. An immunohistochemical study was undertaken on the expression of standard CD44 (CD44s) and the variant isoforms containing the domains encoded by variant exon 3 (CD44v3) or variant exon 6 (CD44v6) in paraffin-embedded bronchial biopsy specimens from 32 NSCLC cases and 11 SCLC cases. An absolute lack of immunoreactivity for CD44s, CD44v3, and CD44v6 was obtained in every case of SCLC, whereas 28 of the 32 NSCLC cases showed a positive immunoreaction forat least oneofthe threeepitopesinvestigated. In conclusion, the occurrence of standard and variant CD44 isoforms in NSCLC and their absence in SCLC suggest the possibility that CD44 is in some way instrumental in conditioning the biological behaviour of NSCLC, but not of SCLC, whose metastatic cascade would be set in motion by the activation of hitherto unidentified, CD44-independent pathways.

Pulmonary myxoid leiomyosarcoma Koizumi N, Fukuda T, Ohnishi Y, Naito M, Emma I, Sato K et al. SecondDepartment ofPathology, Niigata Univ. School of Medicine, 1 Asahimachi dori, Niigata 951. Path01 lnt 1995;45:879-84.

A primary myxoid leiomyosarcoma arising from the peripheral bronchus of the right middle lobe was removed from a 20 year old man and examined by immunohistochemistry and electron microscopy. The tumor was well circumscribed, yellow-whitish focal polypoid growth into the bronchial lumen and consisted of spindle cells with abundant myxoid substance in the stroma. Intrapulmonary metastasis, invasion to the bronchial wall and a few mitotic tigures were found. Immuno- histochemically, several, but not all, tumor cells were positive against anti-vimentin, anti-S-100 protein, anti-myosin and anti-muscle specific actin. Ultrastructurally, tumor cells had thin filaments with dense bodies, pinocytic vesicles and discontinuous basal lamina. These findings indicate a myoxoid variant of leiomyosarcoma arising from mesenchyme in the p&bronchial area.

A rarecaseof primary pulmonary choriocarcinoma in a male: Immunohistochemical detection for human chorionic gonadotropin, epidermal growth factor (EGFl and EGF- receptor Toda S, Inoue Y, Ishino T, Yonemitsu N, Terayama K, Miyabara S et al. Depanme~t of Pathology, Saga Medical School, 5-1-l Nabeshima. Saga-849. Endocr J 1995;42:655-9.

In situ end-labelling, light microscopic assessment and ultrastructure of apoptosis in lung carcinoma Graffney EF, O’Neill AJ, Staunton MJ. DepartmentofHistopathology, St James’s Hospital, Dublin 8. J Clin Path01 1995;48:1017-21.

Aims -To compare in situ end-labelling (ISEL) of apoptosis in lung carcinoma with quantitative and semiquantitative light microscopic assessment and ultrastructural observations. Methods - ISELofapoptosis was evaluated in 42 lung carcinomas (24 squamous cell carcinomas, 12 adenocarcinomas and six small cell carcinomas). Results werecorrelated semiquantitatively with the extent of apoptosis in haematoxylin and eosin stained sections, with apoptotic indices and with ultrastructural observations (nine cases). Results - In each tumour type the extent of apoptosis identified by ISEL correlated with that observed on light and electron microscopy. Tumour cells undergoing apoptosis showed either uniform nuclear staining with a surrounding ‘halo’ or peripheral nuclear membrane staining. The latter pattern was more prominent in small cell carcinoma and correlated ultrastructurally with early apoptosis. A variable proportion of apoptotic cells and apoptotic bodies were unlabelled. Necrotic tumour cells were weakly stained but were distinguishable from apoptotic cells. Conclusions - ISEL, if used in conjunction with standard methods for investigating apoptosis, is a useful adjunct to the investigation of apoptosis in human tumour tissue.

Expression of bcl-2 protein in stage TlNOMO non-small cell lung carcinoma Ritter JH, Dresler CM, Wick MR. Department of Surgical Pathology. Barnes Hospital, Box 8118. One Barnes Hospital Plaza, St. Louis. MO 63110. Hum Pathol 1995;26: 1227-32.

Thebcl-2geneproduct(h&2protein, BCLP) preventsapoptoticcell death. Via a 14;18 chromosomal translocation, BCLP is overexpressed in most follicular lymphomas as well as some other non-Hodgkin’s lymphomas, andithasalsobeendocumentedinothernonlymphomatous malignancies. To address the possible prognostic value of this marker in predefined subsets of non- small cell lung carcinoma (NSCLC), the authorsstudied 126TlNOMOcasesseenbetween the years 1986 to 1991 at our institution. Patients were treated by lobectomy (105 cases) or wedge excision (21 cases) with negative margins; neuroendocrine carcinomas of all grades were specifically excluded. The mean follow- up period was 39 months. Immunostaining for BCLP was done using a monoclonal antibody (clone no. 124; DAKO, Carpinteria, CA), and the avidin- biotin-peroxidase complex (ABC) technique. The study cases included 73 adenocarcinomas (ACs) as well as 40 squamous cell @CC), five adenosquamous (ASC), and eight large cell/poorly differentiated (LCC) carcinomas. As assessed with the Kaplan-Meier method, overall survival was 64% at 5 years (66 % AC vs 59 % SC). BCLP was detected in 47 of 126 cases (37%) including 32 AC (44%), 10 SCC (25%) two ASC (40%), and three LCC (38%). No significant difference in 5-year survival was noted in a comparison of all cases with BCLP expression (63%) and those without (59%). There was, however, a significant difference in the survival of grade 1 BCLP( +) cases, when compared with grade 2 or 3 BCLP(+) cases (P = .Ol). A nonstatistically

Abstracts/kg Gmcer 14 (19%) 377-408

significant trend toward increased survival was observed in BCLP( +) SCC cases (66 96 5-year survival in BCLP[ +] vs 43 46 in BCLP[-] [P = .l I]). Proportional hazards analysis failed to disclose significant independent risk factors. These data suggest that bcl-2 protein immunoreactivity has limited prognostic value in the pathological evaluation of NSCLC.

Theexpression of trophoblasticcell markers by lung carcinomas Boucher LD, Yom& K. PathologyLaboratory Medicine SW. , Veterans Affairs Medical Center, 22SOLeestown Rd. Lexington, KY4051 I-1093. Hum Path01 1995;26:1201-6.

The authors studied the expression of trophoblastic cell markers in lung carcinomas cells by immtmopetoxidase staining using antibodies againstthreetropboblasticglycoproteins(humanchorionicgonadotropin [hCG]; human placental lactogen ]h]PL]; pregnancy-specific 8,- glycoprotein [SP-I]). One hundred five tissue sections from 44 lung carcinomas of various histological types were examined for positive staining with three antibodies. Hematoxylin- rosin-stained sections from the same tissue blocks were examined for the presence of tumor giant cells. The aim was to study the relationship between tumor giant cells and die trophoblaatic glycoprotein expression in lung carcinomas. Small cell carcinoma (XC) did not show any positive reaction with all three markers. Squamous cell carcinoma (SQCC) showed positive staining with hCG in 21% of casea, hPL in 28%, and SP-1 in 64%. Adenocarcinoma showed positive staining with hCG in 60% of cases, hPL in IO%, and SP-1 in 80%. Large cell carcinoma (LCC) showed positive staining with hCG in 93% of cases, hPL in 56%, and SP-I in 93%. The positive reaction did not appear to be restricted to nor associated with the tumor giant cells. It was concluded that these trophoblastic cell markers are expressed in various types of lung carcinomas and that they are not associated with certain histological types or with tumor giant cells.

In situ detection of Epstein-Barr virus in non-small cell lung carcinomas Wong MP, Chung LP, Yuen ST, Leung SY, Chart SY, Wang E et al. Department of Pathology, Queen Mary Hospital, PorgUkam Road, Hong Kong. J Pathol 1995;177:23340.

Epstein-Barr virus (EBV) is strongly associated with nasopharyngeal carcinoma and lymphoepithelioma-like carcinomas (LELC) of foregut- derived organs. Recently this group of EBV-associated carcinomas has been expanded by the identification of the virus in conventional adenocarcinomas of the stomach. In situ hybridization (ISH) using a sensitivedigoxigenin-IabelledEBER RNAprobewt~sperforn~don 167 consecutive unselected primary non-small cell lung carcinomas, to determine the frequency of EBV association in these tumours. Nine cases (5.4 per cent) showed strong EBER signals in the tumour cell nuclei. By immunohistochemistry, four of the EBER-positive tumours showed patchy expression of the viral latent membrane protein (LMP- 1) and none showed any expression of the EBV nuclear antigen 2 (EBNA2). Morphologically, all the positive turnouts were LELC, whereas no conventional type of non-small cell lung carcinoma showed EBV association. The LELC presented a morphological spectrum from undifferentiated to squamoid or glandular differentiation. The patients showed a male to female ratio of 8: 1. The mean age at presentation was 48 years. Smoking was not a risk factor. All patients were alive at follow-up periods of 23-52 months. Southern blot anaiysis performed on eight of the nine positive tumours showed a clonal episomal form of EBV, suggesting the clonal expansion of an infected tumour cell early in oncogenesis. These characteristics of the EBV-associated lung tumours justify their consideration as a distinct clinicopathological entity.

Activation of the precursor of gelatinase A/72 kDa type IV eollagenase/MMP-2 in lung carcinomas correlates with the expression of membrane-type matrix metalloproteinase (MT- MMP) and with lymph node metastasis Tokuraku M, Sato H, Murakami S, Okada Y, Watanabe Y, Seiki M. Dept. Molecular Virology/Oncology. Cancer Research Institute, Kanazawa University, Takara-machi 13-1, Kanazawa. Ishikawa 920. Int J Cancer 1995;64:355-9.

We have identified a novel membrane-type matrix metalloproteinase (MT-MMP) expressed on the cell surface and inducing activation of pro gelatinase A in vitro. In this study, we further examined the possibility that MT-MMP is the activator of pro-gelatinam A in tumors as well as in vitro. Expression of MT-MMP mRNA was analyzed by Northern blotting in 58 cases of human lung carcinomas. MT-MMP mRNA expression was increased in tumor tissues compared with adjacent normal tissues. The ratio of MT-MMP mRNA levels in tumor/normal tissues(T/Nratio) was 3.19 it 1.62 in29casesofadenocarcinoma, 3.09 f 1.44 in 24 cases of squamous cell carcinoma, 4.40 f 0.47 in 3 cases of large cell carcinoma and 3.63 f 2.11 in 2 cases of small cell carcinoma, respectively. Activated gelatinase A, as detected by gelatin zymography, was also predominant in tumors compared with normal tissue counterparts, though the difference in mRNA levels was not significant. The activation ratio of gelatinase A in tumor vs. normal tissues correlated well with that of MT-MMP mRNA expression and with lymph node metastases. Our findings suggest that MT-MMP is indeedthetumor-specificactivatorofpro-gelatinase Am lungcarcinomas and is important to initiate invasion of basement membranes.

ClinicalsignifIcanceorargyruphilicnucleolarorganizerregions (AgNOBs) in squamous cell carcinoma of the lung Han SB, Jean YJ, Lee SS. Depanment of Internal Medicine, Institute of Medical Science, Keimyung Univ. School of Medicine, Taegu. Tuberc Respir Dis 1995; 42:513-21.

Background: Nucleolar organizer regions (NORs) are chromosomal segments encoding for ribosomal RNA and associated with argyrophilic nonhistone protein. Ribosomal RNA genes ultimately direct ribosome and protein synthesis, and it has been suggested the numbers of NORs detected in the cell may reflect nuclear and cellular activity. This study was performed to evaluate the applicability of AgNORs to the diagnosis of squamous cell carcinoma of the lung. Method: The one step silver methods (AgNORs) was used to stain NORs in the routinely processed, formalin fixed, paraffin embedded sections of 36 cases of squamous cell carcinoma of the lung obtained by surgical resection of primary tumor. In each specimen, 100 tumor cells and 100 normal cells adjacent to the tumor chosen at random were examined under an oil immersion lens at a magnification of x 1000. The mean number of AgNORs per nucleus was calculated for each specimen. Results: The mean number of AgNORs per nucleus (mAgNORs) of normal bronchial epithelium and squamous cell carcinomaof the lung was 1.74 + 0.25 and 4.05 * 0.80, respectively. The difference of mAgNOR between normal and tumor tissue was statistically significant (p < 0.001). Therewas no statistical differenceamong tumorsofdifferentstages. ThedifferenceofmAgNOR between normal and tumor tissue was statistically significant in each TNM stage (p < 0.05). Conclusion: Mean AgNOR count may be used as a useful marker for the differential diagnosis of benignancy and malignancy, and proliferative activity of the cell in squamous cell carcinoma of the lung. But there was no statistical difference in mean AgNOR count among tumors of different surgical stages. Further studies for the application of mAgNORs to the diagnosis of other histologic types and cytologic specimens of the lung cancer are needed.