A. Becciolini

Evidence for reciprocity of bcl-2 and p53 expression in human colorectal adenomas and carcinomas

Evidence of accumulating for the failure of apoptosis as an important factor in the evolution of colorectal cancer and its poor response to adjuvant therapy. The proto-oncogene bcl-2 suppresses apoptosis. Its expression could provide an important survival advantage permitting the development of colorectal cancer. The expression of bcl-2 and p53 was determined by immunohistochemistry in 47 samples of histologically normal colonic mucosa, 19 adenomas and 53 adenocarcinomas. Expression of bcl-2 in colonic crypts > 5 cm from the tumours was confined to crypt bases but was more extensive and intense in normal crypts < 5 mm from cancers. A higher proportion of adenomas (63.2%) than carcinomas (36.5%) expressed bcl-2 (P < 0.05). A lower proportion of adenomas (31.6%) than carcinomas (62.3%) expressed p53 (P < 0.02). A total of 26.3% of adenomas and 22% of carcinomas expressed both bcl-2 and p53. To determine whether these samples contained cells which expressed both proteins, a dual staining technique for bcl-2 and p53 was used. Only 1/19 adenomas and 2/53 carcinomas contained cells immunopositive for both bcl-2 and p53. Moreover there was evidence of reciprocity of expression of bcl-2 and p53 in these three double staining neoplasms. We suggest that bcl-2 provides a survival advantage in the proliferative compartment of normal crypts and colorectal neoplasms. However, its expression is lost during the evolution from adenoma to carcinoma, whereas p53 expression is increased, an event generally coincident with the expression of stabilised p53, which we presume to represent the mutant form.

Analysis of HPV16, 18, 31, and 35 DNA in pre-invasive and invasive lesions of the uterine cervix.

AIMS: To analyse the physical state of different human papillomavirus (HPV) DNAs in 55 intraepithelial and invasive HPV associated cervical neoplasms. METHODS: Restriction analysis, using a panel of five HPV type specific enzymes, was carried out for each sample; this was followed by Southern blot analysis. RESULTS: Six (25%) of 24 cervical intraepithelial neoplasms had integrated DNA of different HPV types. In contrast, integration was detected in 25 (81%) of 31 cervical carcinomas. Tumour samples revealed differences in the integration profile of HPV16 and the other HPV types. Six (26%) of 23 HPV16 associated cancers contained only episomal DNA. In contrast, all eight tumours containing HPV18, 31, or 35 revealed integrated DNA exclusively. CONCLUSIONS: The results suggest that in advanced cervical intraepithelial neoplasia lesions, a subset of lesions can be identified in which the viral genome is integrated and there is a greater risk of malignant progression. In addition, HPV16 DNA was not present in the integrated form in 26% of tumours, suggesting that integration and subsequent inactivation of the transcriptional regulator, E2, are not essential steps for the development of HPV16 associated carcinoma. In this respect, the behaviour of HPV16 associated tumours is different from HPV18, 31, and 35 associated tumours, where the viral genome is always present in the integrated form.

Disease-Free Survival Advantage of Adjuvant Cyclophosphamide, Methotrexate, and Fluorouracil in Patients With Node-Negative, Rapidly Proliferating Breast Cancer: A Randomized Multicenter Study

PURPOSE According to one of the most recent key scientific questions concerning the use of biomarkers in clinical trials, we investigated whether node-negative breast cancer patients, defined as high-risk cases on the basis of tumor cell proliferation, could benefit from cyclophosphamide, methotrexate, and fluorouracil (CMF) adjuvant therapy. PATIENTS AND METHODS Two hundred eighty-one patients with negative nodes and rapidly proliferating tumors, defined according to thymidine labeling index (TLI), were randomized to receive six cycles of CMF or no further treatment after surgery +/- radiotherapy. RESULTS The 5-year disease-free survival (DFS) was 83% for patients treated with CMF compared with 72% in the control group (P: =.028). Adjuvant treatment reduced both locoregional and distant metastases. When clinical outcome was analyzed in cell kinetic subgroups characterized according to tertile criteria, compared with patients in the control arm, 5-year DFS was significantly higher after adjuvant CMF in patients with TLI values in the second (78% v 88%, respectively; P: =.037) and third tertiles (58% v 78%, respectively; P: =.024). CONCLUSION The results from this randomized clinical study indicate that patients with node-negative, rapidly proliferating tumors significantly benefit from adjuvant CMF.

Bcl-w expression in colorectal adenocarcinoma.

We have found that the anti-apoptotic Bcl-2 family protein, Bcl-w, was frequently expressed in colorectal adenocarcinomas, with 69/75 showing positive staining with anti-Bcl-w IgG. Adenomas demonstrated a much lower frequency of Bcl-w expression (only 1 of 17), as did adenocarcinomas from other epithelial tissues such as breast (0/8), stomach (1/12) and cervix (0/12). Bcl-w status could be related to the histopathological classification of the tumours, with TNM stage III tumours showing significantly higher levels of expression than tumours of better prognostic grade (at P = 0.009). Those patients with node involvement also had tumours with significantly elevated levels of Bcl-w (at P = 0.02), compared to those which were node-negative. The results suggest that Bcl-w could play a general role in the progression from adenoma to adenocarcinoma in the colorectal epithelium. Currently, more data are being collected to allow us to assess the importance of Bcl-w for disease progression and patient survival.

Prognostic significance of biologic markers in node-negative breast cancer patients : a prospective study

It is generally thought that future advances in the treatment and cure of breast cancer patients will be made possible through a deeper understanding of tumor biology and an improved capability to define the prognosis of each single patient. This will lead to the formulation of new, more selective, and patient-tailored therapies. It is therefore important, when studying potential prognostic factors, to follow methodologic requirements and guidelines which involve the carrying out of prospective studies as confirmatory steps. Repeatedly or recently investigated prognostic markers (tumor size, menopausal status, ER, PgR, 3H thymidine labeling index, c-erbB-2 and p27 expression) were evaluated on a series of 286 prospectively recruited node negative breast cancer patients who underwent loco-regional treatment alone and were closely followed. The individual and relative prognostic contribution of each variable with respect to other factors, as well as their ability to identify node negative patients at risk, were assessed by univariate and multivariate analysis. At a five-year follow-up, only tumor size (p = 0.021) and TLI (p = 0.016) individually proved to be significant prognostic indicators of relapse-free survival. Conversely, p27 expression was not related to RFS and c-erbB-2 expression appeared to have only a short-term effect on patient prognosis. TLI and tumor size, tested in multivariate analysis along with ER and menopausal status, maintained their independent prognostic relevance. The study, performed on a large series of node-negative patients given loco-regional treatment alone, for the first time prospectively recruited, showed the prognostic relevance of TLI and its independence from other clinico-pathologic and biologic factors over a five-year period.

Tumor microvessel density and prognosis in node-negative breast cancer.

Microvessel density (MVD) of breast cancer is widely regarded as a prognostic factor, but results from studies on the most important case series have produced conflicting results. The present study was performed with confirmatory intent to define the prognostic relevance of MVD on a series of 378 node‐negative‐breast‐cancer patients, much larger than any other series previously analyzed. Microvessels were stained using Factor‐VIII antibody and an immunoperoxidase reaction. MVD was determined independently by 2 observers according to Weidners methods. In parallel, cell proliferation was evaluated as S‐phase fraction and determined according to the 3H‐thymidine‐labeling index method (TLI). Estrogen and progesterone receptors were quantitatively assessed using the dextran‐charcoal technique. Tumor MVD varied greatly from tumor to tumor (2 to 232 MV/mm2) and was unrelated to patient age and menopausal status, or to tumor size, histology and steroid‐receptor status. A significant (p = 0.004) but weak inverse correlation (rs = −0.188) was observed with cell proliferation. Univariate analysis using 40 MV/mm2 as cut‐off showed an inverse relation with 5‐year relapse‐free survival (82% vs. 71%, p = 0.018). This finding was limited to very small tumors, slowly proliferating tumors and ER‐negative tumors. Multivariate analysis identified tumor size and TLI, but not MVD, menopausal status or ER as independent prognostic factors. Int. J. Cancer (Pred. Oncol.) 89:74–80, 2000.

Plasma estrogens and androgens in male breast cancer.

Abstract Androstenedione, estrone, estradiol-17β, estriol and testosterone concentrations have been measured by radioimmunoassay in plasma of 17 human males affected by breast cancer. The mean values of estrone, estradiol-17β, and estriol in male breast cancer were significantly higher than in normal controls of comparable age. Androstendione and testosterone were in normal range. Orchidectomy. performed in 3 subjects, decreased the levels of estradiol-17β, but to a lesser extent those of estrone and estriol.

Post-irradiation hyperamylasemia as a biological dosimeter

Serum alpha-amylase was measured before and 24 h after either total body (31 patients) or localized irradiation including the salivary glands (40 patients) or the pancreatic area (22 patients). A significant increase in amylasemia was observed for doses to the parotid glands larger than 0.5 Gy. A sigmoid function of dose was fitted to the data and predicted a maximum amylasemia level for doses larger than 4 Gy and smaller than 10 Gy. The raw data from other published series were adequately described by the same model. However, the confidence limits of the parameters remained wide, because of a considerable interindividual variability. Post-irradiation hyperamylasemia appears to provide a good criterion for triage of accidentally irradiated patients: 24 h after a dose larger than 2 Gy to the parotid glands, 91% of the patients had an amylasemia level higher than 2.5-fold the upper normal value (sensitivity). Conversely, 96% had their serum amylasemia lower than 2.5-fold the upper normal value when dose was smaller than 2 Gy (specificity). However, a retrospective estimation of the absorbed dose (dosimetry) is not likely to be very precise because of the large interindividual variability.

Effect of Ionizing Radiation on the Enzymes of the Intestinal Mucosa of Rats at Different Time Intervals after Abdominal Irradiation

The enzyme content of the brush border of the small intestine was measured after exposure of the abdomen of rats to 500 and 800 R, respectively, of gamma radiation from a cobalt-60 source. Enzyme activities determined between 4 hours and 16 days after exposure indicated that there were modifications in lactase, maltase, invertase, alkaline phosphatase, LAP (leucineaminopeptidase), and total protein content in homogenates of different segments of the small intestine. The first demonstrable effect, occurring between 4 and 36 hours after exposure was an increase in enzyme activity, which was more marked after 500 R than after 800 R. Thereafter the depression in enzyme levels was correlated with the degree of abnormality of the epithelium. Recovery was almost complete at 5 days after 500 R. Although the activity of some of the enzymes had partly recovered at that time after 800 R, full recovery to preexposure values had still not occurred at 11 days after exposure. Histologic damage to the epithelium, particu...

Biochemical and morphological changes in the epithelial cells of the small intestine after irradiation

Rats have been sacrificed after abdominal irradiation with sublethal doses at very close intervals. Observation at light and electron microscope have been carried out on the small intestine of these animals in order to try to explain the simultaneous modifications of leucineaminopeptidase (LAP) activity. In connection with the increase of LAP activity no modifications have been evidenced, at the light microscope, in the differentiated cells of the villus where the membrane digestion process takes place. In the crypt-villus junction we observe at the electron microscope a remarkable increase in the number of ribosomes since the early intervals. Also during the return phase to normal activities the cells of the crypt and of the crypt-villus junction show a temporaneous increase in the amount of ribosomes. Shortly afterwards we observe an increase of LAP activity.