M. Fontanesi

Relationship between cytosol TPS, TPA and cell proliferation

The serological tumor marker tissue polypeptide antigen (TPA) and the more recently identified tissue-specific polypeptide antigen (TPS) have been reported to be indicators of the proliferation rate of the tumor. In the present investigation we compared the cytosol level of the two markers with the proliferative activity of the tumor measured using the 3H-thymidine labelling index. The preliminary results presented here show that higher TLI is associated with lower cytosol levels of both TPA and TPS. TPA and TPS in the cytosol were significantly associated. These findings are in agreement with the previously demonstrated association between high TPA cytosol levels and better prognosis in breast cancer. Further studies are ongoing in order to: 1. confirm these findings in a larger patient series; 2. investigate any possible prognostic indication provided by TPS; 3. evaluate any possible biological meaning of the negative association between TPA/ TPS and TLI in the cytosol of breast cancer.

DNA content in human colon cancer and non-neoplastic adjacent mucosa.

DNA content was determined by flow cytometry in a series of 51 paired fresh tissue samples of primary colorectal carcinomas and the respective non-neoplastic adjacent mucosa in order to assess the relationship between DNA ploidy and the most commonly used prognostic factors. Aneuploidy was observed in 70.6% of the tumors and more than one aneuploid peak was present in 3.9%. Aneuploid tumor frequency was higher in left (93.3%) and right colon (64.7%) cancers than in rectal carcinomas (60.0%), and multiple aneuploid clones were detected more frequently in men than in women and in patients with advanced disease (Dukes stage D). Non-neoplastic mucosa adjacent to aneuploid tumors showed aneuploidy in 4 out of 51 samples (7.8%). The mucosa adjacent to diploid cancers had only diploid characteristics. Polidy did not correlate with histological abnormalities. These findings suggest that DNA content as determined by flow cytometry needs further study with adequate follow-up to evaluate possible correlations with relapse-free and overall survival. Furthermore the aneuploidy of non-neoplastic mucosa provides evidence for a field defect in mucosa adjacent to colorectal cancer and supports the concept that this alteration may be of influence on carcinogenesis.

Localization of mucinous-like carcinoma associated antigen (MCA) in breast pathology: comparison with carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA).

The topographic distribution of a mucinous-like cancer antigen (MCA) recognized by a monoclonal antibody b-12 (MAb b-12) was assessed in benign (38) and malignant (66) breast tissues. The reactivity of MAb b-12 showed a good selectivity for breast tissues, reacting both with normal tissues and breast cancer. The degree of MCA expression was evaluated in the various groups of breast pathology adopting quantitative criteria of assessment. With the criteria of evaluation adopted, strong staining was observed in 71.4% breast carcinomas. The most positive reaction was demonstrated in mucinous carcinoma. MCA distribution in breast tissue was compared with the distribution of two other antigens, CEA and TPA. Reactivity of MAb b-12 was higher than the reactivity shown by the anti-CEA and anti-TPA antibodies.

Demonstration of mucinous-like carcinoma-associated antigen in bone marrow and lymph node biopsies from patients with breast carcinoma.

Two hundred and fifty bone marrow and 140 lymph nodal biopsies were analyzed immunocytochemically, using a mouse monoclonal antibody b-12 (MAb b-12), which reacts with MCA (mucinous-like carcinoma-associated antigen). The presence of MCA in bone marrow specimens was demonstrated in 102 out of 105 (97.1%) breast cancer metastases, 5 out of 8 (62.5%) gastric cancers, 5 out of 6 (83.3%) colon cancers, 3 out of 5 (60%) prostate cancers, 11 out of 26 (42.3%) lung cancers and 25 out of 30 (83.3%) unknown primary cancers, while no positivity to anti-MCA antibody was found in 30 cases of normal bone marrow biopsies, 5 cases of non epithelial malignancies and 30 cases of hemolymphoproliferative disease. Normal lymph nodes and non-epithelial lymph node metastases did not show any reaction to MAb b-12; on the contrary MCA positive staining was observed in 75 out of 75 (100%) lymph nodal metastases in breast cancer. These results suggest that application of MAb b-12 in immunohistochemistry is valid for the detection of bone marrow and lymph nodal micrometastases of epithelial origin.

Plasma and tissue CEA and TPA markers in operable breast cancer. Preliminary results.

CEA and TPA were studied in sera and in histologic specimens of 200 patients with benign (77) or malignant (123) breast pathology. The frequency and expression of the two markers was different in benign and in cancer tissues. Histologic positivity and high levels of circulating markers were observed more frequently in cancer patients than in patients with benign disease. Tissue positivity for the two tumor markers did not always correlate with elevated levels of circulating markers. Positive CEA and TPA incidence was higher in tissue samples than in serum samples. In the breast cancer group, among 33 patients with histologic positivity for CEA, only 5 cases had circulating CEA levels higher than 5 ng/ml; among 91 patients with histologic positivity for TPA, only 45 cases showed circulating levels for TPA higher than 95 U/l. These findings confirm that tumor size, secretory characteristics and vascular supply are factors affecting the achievement of high circulating marker levels. Combined marker measurement in serum and tissues can provide more information about the presence of a given tumor marker. A limited evaluation of the prognostic meaning of the study of combined CEA and TPA in sera and in tissues was carried out during the follow-up of 60 evaluable patients. Only 5 patients had cancer relapses in the first 12 months from surgery; in 2 of 5 patients TPA was positive initially and at the time of recurrence, in serum as well as in tissues. Circulating CEA gave negative findings in all relapsed patients; 2 of them showed weak positivity only in the histologic staining at the time of presentation.

Preliminary study on thymidine labelling index in human non-small cell lung cancer

Sirs, Among patients affected by lung cancer, those with resectable non-small cell lung cancers (NSCLC) have the best prognosis. Various parameters are predictive of the out come of surgical treatment and the most reliable are histological type, tumor grade, nodal status, clinical stage and the general performance status of the patient. Among the novel prognostic indicators the evaluation of proliferative activity is very important, but very little is known about NSCLC growth rate and prognosis. Alama et al. (1) showed that in univariate analysis survival was significantly longer in NSCLC patients with lower 3Hthymidine labelling index (3H-TdR LI) and with stage III and TI-T2 tumors. Silvestrini et al. (2) reported that in stage I NSCLC patients relapse-free survival and overall survival rates were significantly higher for patients with low 3H-TdR LI or squamous cell tumors than for patients with high 3H-TdR LI or non-squamous cell tumors. We evaluated in 27 surgically resected NSCLC the relationship between cell kinetics as assessed by the 3HTdR LI and various clinical, morphological and biological features. 3H-TdR LI was determined on fresh tumour material immediately after surgery as described by Silvestrini et al. (3, 4, 5). The statistical significance of differences was assessed by means of the KruskallWallis test. The patients median age was 63 years, with a range from 49 to 74 years. Twenty-four patients (89%) were males and only 3 (11.1%) females. The 3H-TdR LI evaluated in 27 primary tumors showed a median value of 6.6% (range 0.0 26.0%). No significant increases in 3H-TdRLI values were observed in men, in patients younger than 60 years, or in tumors resected by lobectomy. The relationship between 3H-TdR LI and clinical

CEA, TPA and CA 15.3 in tissue and serum of breast cancer patients.

We are reporting the preliminary results of an ongoing study on tumor marker determination in breast cancer tissue and serum from patients with primary breast cancer. The expression of tumor markers in the tissue was evaluated using immunometric assays in the cytosol as well as immunohistochemical techniques. CEA, TPA and CA 15.3 were determined in a preliminary group of seven patients affected by invasive ductal carcinoma. Tumor markers were measured in serum and in cytosol using commercially available methods (CEA, MKS, SORIN Biomedica, Saluggia, Italy; CA 15.3Cis Diagnostici, Tronzano, Italy; TPA RIA-Kit Byk Gulden, Milan, Italy). The methods were valuated for use in the cytosol matrix using previously reported criteria (l). Tumor markers were determined in the cytosol of neoplastic tissue and normal breast tissue from the same patient collected distantly from the tumor. The immunohistochemical studies wereperformed as previously described (2). CEA, TPA and CA 15.3 were found both in cancer and in normal breast tissue, in agreement with findings reported by other investigators (3,4). The level of tumor markers was higher in breast cancer than in normal tissue in five out of seven cases for CEA, and in all cases for both TPA and CA 15.3(Tab. I). These findings lead to the conclusion that the evaluated tumor markers are expressed both in normal breast tissue and in breast cancer, their expression being probably enhanced as a consequence of neoplastic transformation. Serum concentrations ofCEA, TPA and CA 15.3did not show any significant correlation with the cytosol content (serumCEA/cytosol-CEA r = 0.65; serum-TPA/cytosolTPA, r = 0.026; serum-CA 15.3/cytosol-CA 15.3, r = 0.259), suggesting that the two compartments are probably independent; serum levels are mainly related to tumor bulk, while the cytosol content is possibly related to the phenotypic pattern of the tissue (5). The immunohistochemical distribution of the evaluated tumor markers was compared with their cytosol levels. Immunostaining was more intense in tumors with higher CEA levels in the cytosol. In the case of TPA an opposite trend was shown since less intense immunostaining was found in the samples with higher cytosol TPA. The immunostaining pat-