- Email: [email protected]
693 Neural cell adhesion molecules (NCAM) expression in malignant mesothelioma
Fig. 1. Survival wrves according to expres~on of GS-l-B4 lectin-binding carbohydrate and p53 protein (a) and of Tn antigen and p53 protein (b)
Results: The major findings obtained were summarized in Table 1 and Fig. 1. Conclusion: Prognosis of lung adenocarcinoma patients is able to predicted with high probability by a combination analysis of expression of p53 protein and carbohydrates at least in part.
I
690
~53 and K-ras are mutated in different subtypes bronchioloalveolar carcinoma
of
A. Marchetti, F. Buttitta, G. Bertacca, V. Carnicelli, S. Pellegrini, P. Gaeta, A. Filardo, A. Chella’ , C.A. Angeletti ‘, G. Bevilacqua. Departments of Oncology; ’ Surgery, University of P&a, Piss, /ta/y Bronchioloalveolar carcinomas (BACs) are well-differentiated peripheral adenocarcinomas of the lung which grow along alveolar septa. The existence of BAC as a separate ciinico-pathological entity has been a matter of controversy, mainly because its histogenesis is uncertain and it is not easily distinguishable from conventional lung adenocarcinoma (CLA). Three subtypes of BAC have been described by using histologic and cytologic criteria: mutinous, non-mutinous and sclerosing BAC. The clinical behaviour of BAC appears to be dependent on the histologic subtype. The different morphologic patterns and clinical outcome of the subtypes of BAC suggest that their biological behaviour may be different from one another and from CLA. We have recently shown that k-ras mutations are constant events in mutinous BAC, whereas their frequency in non-mutinous and sclerosing BAC was similar to that observed in CLA. In the present study we have investigated the genomic status of the p53 gene in 36 BACs (10 mutinous, 22 non-mutinous and 4 sclerosing) by PCR-SSCP and sequencing. In 8 cases (22%) we observed p53 mutations. All of the aberrations of p53 were present in non-mutinous and sclerosing subtypes with a frequency (31%) not dissimilar from that reported in CLA. No p53 genetic aberrations were seen in mutinous BACs. We never observed a concomitant alteration of p53 and K-ras genes in the cases examined. Our results suggest that BACs are a heterogeneous group of lung tumors and that the mutinous form, characterized by constant mutations of the K-ras gene and low frequency of ~53 abnormalities, represents a biological entity separate from both the other two BAC types and CLA. Supported by CNR, ACRO n. 96.00591.PF39; AIRC; MURST
I
691
Episialin
(EMA, MUCl) expression
F. Guddo, A.M. Vignola, C. Reina, Respiratoria, CNR, Palermo, Italy
G. Bonsignore.
and metastases. The current study assesses MUCl mucin expression in a retrospective series of AD and SC lung cancer and compares the prognostic significance of this expression with that of other prognostic factors, such as histological type and grade, the presence of node metastases and tumor stage. The expression of episialin is evaluated on paraffin sections of 45 cases of AD and 15 cases of SC, by APAAP immunohistochemical detection, using a monoclonal antibody. The overall proportion of tumor cell showing episialin expression is classified in two groups: 0% to 79% (Low Level Expression-LLE) and 80% to 100% (High Level Expression-HLE) of positive cells. Moreover, the proportion of cells showing apical membrane (A) and overall (0) immunostaining pattern is quantified. Tumors showing LLE and A pattern, and HLE and 0 pattern are correlated to T, N and pathological staging. Episialin is expressed in NSCLC: 67% of AD show more than 80% of tumor cell positive for episialin. being restricted to apical membrane in bronchioloalveolar (BA) carcinoma and in low grade tumor and overall in high grade tumors. On the contrary, 73% of SC has than 80% of tumor cells positive (p 4 0.01). In AD carcinoma, LLE/A pattern are associated to negative N and Stage I (p < 0.03). High Level Expression and Overall pattern episialin immunostaining, in AD tumors of the lung, are associated to node metastases and advanced pathological staging, suggesting a possible role of the molecule in cancer progression.
in lung cancer Lstituto di Fisiopatologia
NSCLC (Non Small Cell Lung Cancer) is a heterogeneous group of cancers consisting mainly of squamous cell (SC), adeno (AD) and large cell carcinoma. Hitherto, using conventional prognostic factors, such as Tumor Node Metasteses (TNM) stage, great differences in outcome after therapy are seen among patients with the same initial stage, suggesting the need of a better understanding of cancer biological events. Episialin (EMA, PEM), the product of the gene MUCl, is a heavily glycosilated protein. While in glandular epithelial ceil it is present at apical surface, in carcinomas cell polarization is lost, being expressed on the entire cell surface. Overexpression of episialin in vitro reduces cell-cell and cell-matrix adhesion, by masking membrane adhesion receptors. Episialin also exerts its anti-adhesion effect in vivo. As a consequence of its “anti-adhesion” properties, episialin expression on carcinomas cell could be associated to a major “anti-adhesive” effect, with a possible role in tumoral invasion
692
El
Comparison of the expression of VEGF/Flk-1 and bFGF/FGFR-1 in NSCLC and in the surrounding non-neoplastic pulmonary parenchyma
F. Guddo, C. Reina, A.M. Vignola, G. Bonsignore. Respiratoria, CNR, Palermo, /ta/y
lstituto
di Fisiopatologia
Several studies have demonstrated the strict relationship between intratumoral microvessel count (MC) and tumor growth and metastasis in non small cell lung cancer (NSCLC). Neverthless very little is known about the causes determining increased MC in lung cancer. Vascular endothelial growth factor (VEGF) and basic Fibroblast growth factor (bFGF) and their endothelial cell receptors fetal liver kinase-1 (Flk-1), c-fms-like tyrosin kinase-I (Flt-I) and fibroblast growth factor receptor (FGFR-I) play an important role in blood microvessel growth in tumors. Aim of the study was to correlate the expression of VEGF/Flk-1, Flt-1 and FGF/FGFR-1 to MC in NSCLC. The expression of VEGF/Flk-1, Fit-1 and FGFIFGFR-1 was evaluated on paraffin sections in 30 NSCLC (Tl-2, NO-l, MO) by streptavidin-peroxidase detection, using monoclonal and polyclonal antibodies; a polyclonal antibody directed against von Willebrand factor (FVIII) was used to identify microvessels. Proportion of epithelial cells immunoreactive for VEGF/Flk-1, Flt-1 and FGFIFGFR-I and proportion of microvessels immunoreactive for Flk-1, Flt-I and FGFR-1 was assessed both in neoplasia and in surrounding non-neoplastic pulmonary parenchyma. MC is assessed in x.250 microscope fields measured in the area of highest concentration. The expression of VEGF (U Mann-Whitney p = 0.0001) and bFGF (p = 0.0003) was higher in neoplasia than in surrounding non-neoplastic pulmonary parenchyma and MC was related to the expression of bFGF in tumor (p = 0.05). Moreover, Flk-I (p = 0.02) and FGFR-1 (0.0001) were more expressed in epithelial neoplastic cells: MC was related to the proportion of microvessels immunoreactive for Flk-1 both in neoplasia (p = 0.05) and in surrounding non-neoplastic pulmonary parenchyma (p = 0.009). These results show that l-the expression of VEGF and bFGF and of their receptors Flk-1 and FGFR-1 is higher in neoplasia than in surrounding non-neoplastic pulmonary, parenchyma, 2-MC is associated to bFGF expression in neoplasia and 3-MC is associated to the endothelial expression of Flk-1 both in neoplasia and in surrounding non-neoplastic pulmonary parenchyma.
693
El
Neural cell adhesion molecules in malignant mesothelioma
S. Lantubjoul, de Pathologic
(NCAM) expression
H. Sartelet, PY. Brichon, C. Brambilla, Cellulaire, BP 217X-Grenoble, France
E. Brambilla.
Service
NCAM are adhesion molecules expressed by neural and neuroendocrine tumors, but also in a few mesenchymal and biphasic tumors such as synovialosarcoma and breast phylloides tumors. Their expression has never been examined in mesothelioma. We have studied expression of NCAM molecules in 20 cases of mesothelioma including 9 epithelial, IO biphasic and one with sarcoma-
178
Biology
tous pattern. Twenty-five adenocarcinoma (ADC) and 25 squamous cell carcinoma (SCC) were also studied. We investigated NCAM expression using immunohistochemistry on both frozen and formalin fixed tissues, with 123C3 antibody. Comparison was made with others neuroendccrine markers such as chromogranin A and synaptophysin, and electron microscopy was used as a gold standard. At least one malignant component expressed NCAM in 14/20 mesothelioma. We demonstrated a membranar expression of NCAM molecules in 10 cases, within the fusiform component of biphasic and sarcomatous mesothelioma. In 3 epithelial and 3 biphasic mesothelioma, epithelial components were positive in IO to 70% of the cells. Moreover, in 4 epihelial mesothelioma, 10 to 30% of stromal fibroblasts exhibited a strong focal positivity in the vicinity of tumor cells. Chromogranin expression was never seen, whereas synaptophysin was noticed in 3 cases beside NCAM expression. Neither ADC nor SCC were positive for these markers, We conclude that original expression of NCAM in mesothelioma is in keeping with that reported in other biphasic tumors. This has to be taken into account in the differential diagnosis between primary mesothelioma, and non small cell carcinoma (SCC, ADC and neuroendocrine carcinoma) metastatic to the pleura, and between mesothelioma and pleural sarcoma.
I
694
A combination study of P53 protein expression DNA ploidy on differential diagnosis between atypical metaplasia and intraepithelial cancer-spreading in early bronchial carcinoma
and
T. Ogata, Y. Inage, T. Yamamoto, E. Akaogi, H. Horiguchi, H. Kamma. lbaraki Prefectural Univ. of Health Sciences & Tsukuba Univ. Ibaraki, Japan It is important for surgical pathologists to differentiate intraepithelial cancerspreading from atypical metaplasia of the bronchial epithelium at histologic study of biopsies or surgical stumps of the bronchus. A combination study of p53 protein (P53) expression and DNA ploidy was done on the bronchial specimens in cases with early bronchial carcinoma limited to its bronchial extension. Twenty-six lesions of atypical metaplasia and 43 early bronchial carcinomas including 23 carcinomas in situ (CIS) or microinvasive carcinomas were examined by immunohistochemistry of P53 and image cytometry of DNA contents. Cancer cells of the primary lesion and its intraepithelial spreading areas showed similar P53 stain intensity and DNA ploidy. Therefore we made diagnosis as metaplasia, when atypical cells in the bronchial epithelium showed different P53 stain intensity and DNA ploidy from those of primary cancer lesions. Cancer cells were unexpectedly widespread to the adjacent bronchial epithelium even in early bronchial carcinoma. Nuclear P53 was overexpressed in some metaplastic lesions with severe atypia. DNA aneuploidy was only found in cancer cells. Frequency of P53 overexpression and DNA aneuploidy is as follows: P53 OE Nonal spithelium (17) SM without atypia (24) SM with mild atypia (14) SM with savers atypia (12) CISlmicroinvasiva carcinoma (23) SM: squamous metaplasia, 0; number of
DNA AP
0% 0% 0% 0% 76% lesions, OK; overexpression,
0% 0% 0% 25% 64%
AP; aneuploidy
The combination study may be practically useful to differentiate intraepithelial spreading of cancer cells from atypical squamous metaplasia in the bronchial specimens.
I
695
thickening nuclear margin. Recently we developed an image cytometry system using a color image analyzer to calculate geometric parameters (nuclear size, form factor, polymorphic factor and chromatin texture), and to estimate nuclear DNA ploidy and DNA synthetic phase fraction (SPF) simultaneously. We present here the correlation between the geometric parameters and DNA status (ploidy or SPF). Touch smear specimens from 36 non-small cell lung cancers (18 adenocarcinomas, 14 squamous cell carcinomas, two large cell carcinomas, and two carcinoids) were used for this study. We calculated geometric parameters and DNA contents of feulgenstained cancer cells by our image cytometry system. Thirty-six lung cancers were divided into two groups by a DNA index (DI = 1.3) (11 diploidy and 25 aneuploidy cases) and to two groups by a SPF value (SPF = 20%) (16 non-proliferative and 20 proliferative cases). DNA ploidy status was significantly correlated with three geometric parameters of nuclear size, form factor and polymorphic factor, while there was no difference in these geometric parameters between non-proliferative and proliferative groups of cancers. Furthermore, parameters of chromatin texture were significantly different between diploidy and aneuploidy groups of cancers, while there was no difference between non-proliferative and proliferative groups of cancers. In conclusion, morphologic abnormalities of cancer cells may be greatly influenced by DNA aneuploidy of the cells not but by their proliferative activity in NSCLC.
Nuclear abnormalities of cancer cells in morphology are greatly influenced by DNA ploidy but not by proliferative activity in non-small cell lung cancer (NSCLC)
T. Yamamoto, M. Noro, H. Horiguchi, H. Kamma, T. Ogata, M. Matsui, E. Akaogi, S. Ishikawa, K. Mitsui, T. Mitsui. Konan Hosp., Tsukuba Univ. & tbaraki Prefectural Univ. of Health Sciences, Ibaraki, Japan Nuclear abnormalities of cancer cells in morphology have been a diagnostic marker to differentiate cancer cells from normal cells. The abnormalities include enlarged nuclear size, polymorphism, chromatin aggregation, and
I
696
lmmunohistochemical detection of topoisomerase II a (Topolla) P-glycoprotein (Pgp), and multidrug resistance protein (MRP) in non small cell lung cancer (NSCLC) and in small cell lung cancer (SCLC) before and after treatment with topoll directed drugs
J. Kreisholt, M. Sorensen, M. Sehested. Laboratory Copenhagen, Denmark
P.B. Jensen, B.S. Nielsen, C.B. Andersen, and finsen Centres, Rigshospitalef, DK-2100
NSCLC and SCLC differ significantly in their clinical response to topolla directed drugs as etoposide and teniposide as NSCLC is virtually insensitive to single agent therapy, while SCLC responds in 2/3 of cases. Immunohistochemical analysis on paraffin-embedded tissue from 26 cases of untreated NSCLC, and 29 oases of untreated SCLC (10 with additional tumor biopsies after treatment with topoll drugs) gave the Results: NSCLC had significantly less topoll than SCLC as only 6126 NSCLC cases had r5% positive cells compared to 28/29 SCLC, and O/26 NSCLC had >25% positive cells compared to 26/29 SCLC. 7/10 SCLC cases showed a decrease in topoll after treatment. Pgp was detected in >5% of cells in only 3/26 NSCLC and in 6/29 SCLC, and MRP in 8/26 of NSCLC and 9/29 SCLC. After treatment there was an increase in MRP positivity in 4/10 and a decrease in l/10 cases of SCLC while an increase in Pgp was detected in 6/10 patients. In 2/10 cases both MRP and Pgp increased. In Conclusion: The major difference between untreated NSCLC and SCLC was in topoll alpha content. In treated SCLC, emergence of all 3 MDR phenotypes was detected.
0697
p53 expression and gene mutation in operable non-small cell lung cancer (NSCLC)
J. Niklinski, L. Chyczewski, M.A. Sipowicz’, M. Furman. Sialysfok Med. Univ, Balystok, Institute, Frederic, MD, USA
Y.-H. Shiao ‘, J. Laudanski, Poland, ’ National Cancer
We have examined p53 protein expression and gene mutations in 74 surgically treated NSCLC patients, and association of the p53 alterations with the clinicopathological and prognostic parameters. Mutations in exons 5 through 8 of the p53 gene were screened by single strand confirmation polymorphism (SSCP) and DNA sequencing. Accumulation of p53 protein was detected in paraffin-embedded tissue by immunostaining using MAb-DO7 (Dako). Overexpression of ~53, indicated by 210% of tumour cells positive after immunohistochemistry, occurred in 41 of 74 tumours (55%) whereas p53 mutations were found in 19 of 54 tumours (35%). The concordance rate between p53 expression and mutations was 75%. Gene mutations and/or accumulation of ~53 protein were common in both squamous cell (66%) and large cell (66%) carcinomas but significantly less
Results: The major findings obtained were summarized in Table 1 and Fig. 1. Conclusion: Prognosis of lung adenocarcinoma patients is able to predicted with high probability by a combination analysis of expression of p53 protein and carbohydrates at least in part.
I
690
~53 and K-ras are mutated in different subtypes bronchioloalveolar carcinoma
of
A. Marchetti, F. Buttitta, G. Bertacca, V. Carnicelli, S. Pellegrini, P. Gaeta, A. Filardo, A. Chella’ , C.A. Angeletti ‘, G. Bevilacqua. Departments of Oncology; ’ Surgery, University of P&a, Piss, /ta/y Bronchioloalveolar carcinomas (BACs) are well-differentiated peripheral adenocarcinomas of the lung which grow along alveolar septa. The existence of BAC as a separate ciinico-pathological entity has been a matter of controversy, mainly because its histogenesis is uncertain and it is not easily distinguishable from conventional lung adenocarcinoma (CLA). Three subtypes of BAC have been described by using histologic and cytologic criteria: mutinous, non-mutinous and sclerosing BAC. The clinical behaviour of BAC appears to be dependent on the histologic subtype. The different morphologic patterns and clinical outcome of the subtypes of BAC suggest that their biological behaviour may be different from one another and from CLA. We have recently shown that k-ras mutations are constant events in mutinous BAC, whereas their frequency in non-mutinous and sclerosing BAC was similar to that observed in CLA. In the present study we have investigated the genomic status of the p53 gene in 36 BACs (10 mutinous, 22 non-mutinous and 4 sclerosing) by PCR-SSCP and sequencing. In 8 cases (22%) we observed p53 mutations. All of the aberrations of p53 were present in non-mutinous and sclerosing subtypes with a frequency (31%) not dissimilar from that reported in CLA. No p53 genetic aberrations were seen in mutinous BACs. We never observed a concomitant alteration of p53 and K-ras genes in the cases examined. Our results suggest that BACs are a heterogeneous group of lung tumors and that the mutinous form, characterized by constant mutations of the K-ras gene and low frequency of ~53 abnormalities, represents a biological entity separate from both the other two BAC types and CLA. Supported by CNR, ACRO n. 96.00591.PF39; AIRC; MURST
I
691
Episialin
(EMA, MUCl) expression
F. Guddo, A.M. Vignola, C. Reina, Respiratoria, CNR, Palermo, Italy
G. Bonsignore.
and metastases. The current study assesses MUCl mucin expression in a retrospective series of AD and SC lung cancer and compares the prognostic significance of this expression with that of other prognostic factors, such as histological type and grade, the presence of node metastases and tumor stage. The expression of episialin is evaluated on paraffin sections of 45 cases of AD and 15 cases of SC, by APAAP immunohistochemical detection, using a monoclonal antibody. The overall proportion of tumor cell showing episialin expression is classified in two groups: 0% to 79% (Low Level Expression-LLE) and 80% to 100% (High Level Expression-HLE) of positive cells. Moreover, the proportion of cells showing apical membrane (A) and overall (0) immunostaining pattern is quantified. Tumors showing LLE and A pattern, and HLE and 0 pattern are correlated to T, N and pathological staging. Episialin is expressed in NSCLC: 67% of AD show more than 80% of tumor cell positive for episialin. being restricted to apical membrane in bronchioloalveolar (BA) carcinoma and in low grade tumor and overall in high grade tumors. On the contrary, 73% of SC has than 80% of tumor cells positive (p 4 0.01). In AD carcinoma, LLE/A pattern are associated to negative N and Stage I (p < 0.03). High Level Expression and Overall pattern episialin immunostaining, in AD tumors of the lung, are associated to node metastases and advanced pathological staging, suggesting a possible role of the molecule in cancer progression.
in lung cancer Lstituto di Fisiopatologia
NSCLC (Non Small Cell Lung Cancer) is a heterogeneous group of cancers consisting mainly of squamous cell (SC), adeno (AD) and large cell carcinoma. Hitherto, using conventional prognostic factors, such as Tumor Node Metasteses (TNM) stage, great differences in outcome after therapy are seen among patients with the same initial stage, suggesting the need of a better understanding of cancer biological events. Episialin (EMA, PEM), the product of the gene MUCl, is a heavily glycosilated protein. While in glandular epithelial ceil it is present at apical surface, in carcinomas cell polarization is lost, being expressed on the entire cell surface. Overexpression of episialin in vitro reduces cell-cell and cell-matrix adhesion, by masking membrane adhesion receptors. Episialin also exerts its anti-adhesion effect in vivo. As a consequence of its “anti-adhesion” properties, episialin expression on carcinomas cell could be associated to a major “anti-adhesive” effect, with a possible role in tumoral invasion
692
El
Comparison of the expression of VEGF/Flk-1 and bFGF/FGFR-1 in NSCLC and in the surrounding non-neoplastic pulmonary parenchyma
F. Guddo, C. Reina, A.M. Vignola, G. Bonsignore. Respiratoria, CNR, Palermo, /ta/y
lstituto
di Fisiopatologia
Several studies have demonstrated the strict relationship between intratumoral microvessel count (MC) and tumor growth and metastasis in non small cell lung cancer (NSCLC). Neverthless very little is known about the causes determining increased MC in lung cancer. Vascular endothelial growth factor (VEGF) and basic Fibroblast growth factor (bFGF) and their endothelial cell receptors fetal liver kinase-1 (Flk-1), c-fms-like tyrosin kinase-I (Flt-I) and fibroblast growth factor receptor (FGFR-I) play an important role in blood microvessel growth in tumors. Aim of the study was to correlate the expression of VEGF/Flk-1, Flt-1 and FGF/FGFR-1 to MC in NSCLC. The expression of VEGF/Flk-1, Fit-1 and FGFIFGFR-1 was evaluated on paraffin sections in 30 NSCLC (Tl-2, NO-l, MO) by streptavidin-peroxidase detection, using monoclonal and polyclonal antibodies; a polyclonal antibody directed against von Willebrand factor (FVIII) was used to identify microvessels. Proportion of epithelial cells immunoreactive for VEGF/Flk-1, Flt-1 and FGFIFGFR-I and proportion of microvessels immunoreactive for Flk-1, Flt-I and FGFR-1 was assessed both in neoplasia and in surrounding non-neoplastic pulmonary parenchyma. MC is assessed in x.250 microscope fields measured in the area of highest concentration. The expression of VEGF (U Mann-Whitney p = 0.0001) and bFGF (p = 0.0003) was higher in neoplasia than in surrounding non-neoplastic pulmonary parenchyma and MC was related to the expression of bFGF in tumor (p = 0.05). Moreover, Flk-I (p = 0.02) and FGFR-1 (0.0001) were more expressed in epithelial neoplastic cells: MC was related to the proportion of microvessels immunoreactive for Flk-1 both in neoplasia (p = 0.05) and in surrounding non-neoplastic pulmonary parenchyma (p = 0.009). These results show that l-the expression of VEGF and bFGF and of their receptors Flk-1 and FGFR-1 is higher in neoplasia than in surrounding non-neoplastic pulmonary, parenchyma, 2-MC is associated to bFGF expression in neoplasia and 3-MC is associated to the endothelial expression of Flk-1 both in neoplasia and in surrounding non-neoplastic pulmonary parenchyma.
693
El
Neural cell adhesion molecules in malignant mesothelioma
S. Lantubjoul, de Pathologic
(NCAM) expression
H. Sartelet, PY. Brichon, C. Brambilla, Cellulaire, BP 217X-Grenoble, France
E. Brambilla.
Service
NCAM are adhesion molecules expressed by neural and neuroendocrine tumors, but also in a few mesenchymal and biphasic tumors such as synovialosarcoma and breast phylloides tumors. Their expression has never been examined in mesothelioma. We have studied expression of NCAM molecules in 20 cases of mesothelioma including 9 epithelial, IO biphasic and one with sarcoma-
178
Biology
tous pattern. Twenty-five adenocarcinoma (ADC) and 25 squamous cell carcinoma (SCC) were also studied. We investigated NCAM expression using immunohistochemistry on both frozen and formalin fixed tissues, with 123C3 antibody. Comparison was made with others neuroendccrine markers such as chromogranin A and synaptophysin, and electron microscopy was used as a gold standard. At least one malignant component expressed NCAM in 14/20 mesothelioma. We demonstrated a membranar expression of NCAM molecules in 10 cases, within the fusiform component of biphasic and sarcomatous mesothelioma. In 3 epithelial and 3 biphasic mesothelioma, epithelial components were positive in IO to 70% of the cells. Moreover, in 4 epihelial mesothelioma, 10 to 30% of stromal fibroblasts exhibited a strong focal positivity in the vicinity of tumor cells. Chromogranin expression was never seen, whereas synaptophysin was noticed in 3 cases beside NCAM expression. Neither ADC nor SCC were positive for these markers, We conclude that original expression of NCAM in mesothelioma is in keeping with that reported in other biphasic tumors. This has to be taken into account in the differential diagnosis between primary mesothelioma, and non small cell carcinoma (SCC, ADC and neuroendocrine carcinoma) metastatic to the pleura, and between mesothelioma and pleural sarcoma.
I
694
A combination study of P53 protein expression DNA ploidy on differential diagnosis between atypical metaplasia and intraepithelial cancer-spreading in early bronchial carcinoma
and
T. Ogata, Y. Inage, T. Yamamoto, E. Akaogi, H. Horiguchi, H. Kamma. lbaraki Prefectural Univ. of Health Sciences & Tsukuba Univ. Ibaraki, Japan It is important for surgical pathologists to differentiate intraepithelial cancerspreading from atypical metaplasia of the bronchial epithelium at histologic study of biopsies or surgical stumps of the bronchus. A combination study of p53 protein (P53) expression and DNA ploidy was done on the bronchial specimens in cases with early bronchial carcinoma limited to its bronchial extension. Twenty-six lesions of atypical metaplasia and 43 early bronchial carcinomas including 23 carcinomas in situ (CIS) or microinvasive carcinomas were examined by immunohistochemistry of P53 and image cytometry of DNA contents. Cancer cells of the primary lesion and its intraepithelial spreading areas showed similar P53 stain intensity and DNA ploidy. Therefore we made diagnosis as metaplasia, when atypical cells in the bronchial epithelium showed different P53 stain intensity and DNA ploidy from those of primary cancer lesions. Cancer cells were unexpectedly widespread to the adjacent bronchial epithelium even in early bronchial carcinoma. Nuclear P53 was overexpressed in some metaplastic lesions with severe atypia. DNA aneuploidy was only found in cancer cells. Frequency of P53 overexpression and DNA aneuploidy is as follows: P53 OE Nonal spithelium (17) SM without atypia (24) SM with mild atypia (14) SM with savers atypia (12) CISlmicroinvasiva carcinoma (23) SM: squamous metaplasia, 0; number of
DNA AP
0% 0% 0% 0% 76% lesions, OK; overexpression,
0% 0% 0% 25% 64%
AP; aneuploidy
The combination study may be practically useful to differentiate intraepithelial spreading of cancer cells from atypical squamous metaplasia in the bronchial specimens.
I
695
thickening nuclear margin. Recently we developed an image cytometry system using a color image analyzer to calculate geometric parameters (nuclear size, form factor, polymorphic factor and chromatin texture), and to estimate nuclear DNA ploidy and DNA synthetic phase fraction (SPF) simultaneously. We present here the correlation between the geometric parameters and DNA status (ploidy or SPF). Touch smear specimens from 36 non-small cell lung cancers (18 adenocarcinomas, 14 squamous cell carcinomas, two large cell carcinomas, and two carcinoids) were used for this study. We calculated geometric parameters and DNA contents of feulgenstained cancer cells by our image cytometry system. Thirty-six lung cancers were divided into two groups by a DNA index (DI = 1.3) (11 diploidy and 25 aneuploidy cases) and to two groups by a SPF value (SPF = 20%) (16 non-proliferative and 20 proliferative cases). DNA ploidy status was significantly correlated with three geometric parameters of nuclear size, form factor and polymorphic factor, while there was no difference in these geometric parameters between non-proliferative and proliferative groups of cancers. Furthermore, parameters of chromatin texture were significantly different between diploidy and aneuploidy groups of cancers, while there was no difference between non-proliferative and proliferative groups of cancers. In conclusion, morphologic abnormalities of cancer cells may be greatly influenced by DNA aneuploidy of the cells not but by their proliferative activity in NSCLC.
Nuclear abnormalities of cancer cells in morphology are greatly influenced by DNA ploidy but not by proliferative activity in non-small cell lung cancer (NSCLC)
T. Yamamoto, M. Noro, H. Horiguchi, H. Kamma, T. Ogata, M. Matsui, E. Akaogi, S. Ishikawa, K. Mitsui, T. Mitsui. Konan Hosp., Tsukuba Univ. & tbaraki Prefectural Univ. of Health Sciences, Ibaraki, Japan Nuclear abnormalities of cancer cells in morphology have been a diagnostic marker to differentiate cancer cells from normal cells. The abnormalities include enlarged nuclear size, polymorphism, chromatin aggregation, and
I
696
lmmunohistochemical detection of topoisomerase II a (Topolla) P-glycoprotein (Pgp), and multidrug resistance protein (MRP) in non small cell lung cancer (NSCLC) and in small cell lung cancer (SCLC) before and after treatment with topoll directed drugs
J. Kreisholt, M. Sorensen, M. Sehested. Laboratory Copenhagen, Denmark
P.B. Jensen, B.S. Nielsen, C.B. Andersen, and finsen Centres, Rigshospitalef, DK-2100
NSCLC and SCLC differ significantly in their clinical response to topolla directed drugs as etoposide and teniposide as NSCLC is virtually insensitive to single agent therapy, while SCLC responds in 2/3 of cases. Immunohistochemical analysis on paraffin-embedded tissue from 26 cases of untreated NSCLC, and 29 oases of untreated SCLC (10 with additional tumor biopsies after treatment with topoll drugs) gave the Results: NSCLC had significantly less topoll than SCLC as only 6126 NSCLC cases had r5% positive cells compared to 28/29 SCLC, and O/26 NSCLC had >25% positive cells compared to 26/29 SCLC. 7/10 SCLC cases showed a decrease in topoll after treatment. Pgp was detected in >5% of cells in only 3/26 NSCLC and in 6/29 SCLC, and MRP in 8/26 of NSCLC and 9/29 SCLC. After treatment there was an increase in MRP positivity in 4/10 and a decrease in l/10 cases of SCLC while an increase in Pgp was detected in 6/10 patients. In 2/10 cases both MRP and Pgp increased. In Conclusion: The major difference between untreated NSCLC and SCLC was in topoll alpha content. In treated SCLC, emergence of all 3 MDR phenotypes was detected.
0697
p53 expression and gene mutation in operable non-small cell lung cancer (NSCLC)
J. Niklinski, L. Chyczewski, M.A. Sipowicz’, M. Furman. Sialysfok Med. Univ, Balystok, Institute, Frederic, MD, USA
Y.-H. Shiao ‘, J. Laudanski, Poland, ’ National Cancer
We have examined p53 protein expression and gene mutations in 74 surgically treated NSCLC patients, and association of the p53 alterations with the clinicopathological and prognostic parameters. Mutations in exons 5 through 8 of the p53 gene were screened by single strand confirmation polymorphism (SSCP) and DNA sequencing. Accumulation of p53 protein was detected in paraffin-embedded tissue by immunostaining using MAb-DO7 (Dako). Overexpression of ~53, indicated by 210% of tumour cells positive after immunohistochemistry, occurred in 41 of 74 tumours (55%) whereas p53 mutations were found in 19 of 54 tumours (35%). The concordance rate between p53 expression and mutations was 75%. Gene mutations and/or accumulation of ~53 protein were common in both squamous cell (66%) and large cell (66%) carcinomas but significantly less