J. Schneider

Elevated mammaglobin (h-MAM) expression in breast cancer is associated with clinical and biological features defining a less aggressive tumour phenotype.

BackgroundMammaglobin (h-MAM) is expressed mainly by breast epithelial cells, and this feature has been used to detect circulating breast cancer cells and occult metastases in sentinel axillary lymph nodes of breast cancer patients. However, the biological role of mammaglobin is completely unknown.MethodsWe studied 128 fresh-frozen breast cancer specimens by means of reverse transcriptase–polymerase chain reaction and quantified their h-MAM mRNA expression. This was then correlated with histological and nuclear grade, oestrogen and progesterone receptor expression, c-erb-B2 and mutant p53 expression, as well as with cellular proliferation measured by means of the Ki67 labelling index, DNA ploidy and S-phase, and finally with the presence or not of invaded axillary nodes in the mastectomy specimen.ResultsIn the univariate analysis, high h-MAM expression (above the median for the whole group) correlated significantly (P < 0.05) with oestrogen and progesterone receptor expression, diploid DNA content, low Ki67 labelling index, low nuclear grade and almost significantly (P = 0.058) with the absence of axillary nodal invasion in the mastectomy specimen. In a final, multivariate model, only progesterone receptor expression, diploid DNA content and absence of nodal invasion were found to be independently associated with high h-MAM expression.ConclusionAll of the features associated with mammaglobin expression reflect, without exception, a less aggressive tumour phenotype. Further studies are needed to clarify whether this is attributable to h-MAM expression itself, or to another mechanism of which mammaglobin expression forms part.

A Novel, Nuclear Pore-Associated, Widely Distributed Molecule Overexpressed in Oncogenesis and Development

Nuclear pore complexes are large, elaborate macromolecular structures that mediate the bidirectional nucleocytoplasmic traffic. In vertebrates, nuclear pore complexes comprise 50 to 100 proteins termed nucleoporins (Nup). An 88-kd nucleoporin (Nup88) has been recently cloned and characterized, and found to be associated in a dynamic subcomplex with the oncogenic nucleoporin CAN/Nup 214. We have produced a polyclonal antiserum to Nup88, and found that it immunoreacts convincingly in conventional tissue sections of 214 samples of malignant tumors of many types. All carcinomas were stained irrespective of site or line of differentiation; the majority of cases reacted strongly and extensively. In situ carcinomas and highly dysplastic epithelia were similarly reactive. Samples of malignant mesotheliomas, gliomas, sarcomas, and lymphoreticular tumors were also stained. Substantial reactions were also found in certain fetal tissues. Focal reactions were noted in some reactive-proliferative processes. Most benign epithelial and mesenchymal tumors and hyperplasias, and normal adult tissues reacted weakly and sporadically or not at all. Immunoblot analysis of selected samples strongly corroborated those findings. If further substantiated, our findings indicate that Nup88 could be regarded as a selective yet broadly based proliferation marker of potential significance in the histological evaluation and diagnosis of malignant transformation. Its ready applicability on conventional paraffin sections and on cytological preparations may broaden its clinical and investigative significance.

Expression of LRP and MDR1 in locally advanced breast cancer predicts axillary node invasion at the time of rescue mastectomy after induction chemotherapy

BackgroundAxillary node status after induction chemotherapy for locally advanced breast cancer has been shown on multivariate analysis to be an independent predictor of relapse. However, it has been postulated that responders to induction chemotherapy with a clinically negative axilla could be spared the burden of lymphadenectomy, because most of them will not show histological nodal invasion. P-glycoprotein expression in the rescue mastectomy specimen has finally been identified as a significant predictor of patient survival.MethodsWe studied the expression of the genes encoding multidrug resistance associated protein (MDR1) and lung cancer associated resistance protein (LRP) in formalin-fixed, paraffin-embedded tumor samples from 52 patients treated for locally advanced breast cancer by means of induction chemotherapy followed by rescue mastectomy. P-glycoprotein expression was assessed by means of immunohistochemistry before treatment in 23 cases, and by means of reverse-transcriptase-mediated polymerase chain reaction (RT-PCR) after treatment in 46 (6 failed). LRP expression was detected by means of immunohistochemistry, with the LRP-56 monoclonal antibody, in 31 cases before treatment. Immunohistochemistry for detecting the expression of c-erb-B2, p53, Ki67, estrogen receptor and progesterone receptor are routinely performed in our laboratory in every case, and the results obtained were included in the study. All patients had received between two and six cycles of standard 5-fluorouracil, doxorubicin and cyclophosphamide (FAC) chemotherapy, with two exceptions [one patient received four cycles of a docetaxel-adriamycin combination, and the other four cycles of standard cyclophosphamide-methotrexate-5-fluorouracil (CMF) polychemotherapy]. Response was assessed in accordance with the Response Evaluation Criteria In Solid Tumors (RECIST). By these, 2 patients achieved a complete clinical response, 37 a partial response, and the remaining 13 showed stable disease. This makes a total clinical response rate of 75.0%. None achieved a complete pathological response.ResultsMDR1 mRNA expression detected by RT-PCR was associated with the presence of invaded axillary nodes at surgery in 18/22 cases (81.8%), compared with 13/24 (54.2%) in the group with undetectable MDR1 expression. This difference was statistically significant (P < 0.05). LRP expression in more than 20% of tumor cells before any treatment was associated with axillary nodal metastasis after chemotherapy and rescue mastectomy in 17/23 cases, compared with 3/8 in nonexpressors. Again, this difference was highly significant (P < 0.01). LRP expression before treatment and MDR1 mRNA expression after treatment were significantly interrelated (P < 0.001), which might reflect the presence of chemoresistant clones liable to metastasize to the regional nodes. Persistence of previously detected MDR1-positivity after treatment (7/9 compared with 0/2 cases) was significantly associated with axillary node metastasis (P < 0.05). Finally, in a logistic regression multivariate model, histology other than ductal, a Ki67 labeling index of at least 20% and the combination of LRP and MDR1 positivity emerged as independent predictors of axillary node invasion at the time of rescue mastectomy.ConclusionThe expression of different genes involved in resistance to chemotherapy, both before and after treatment with neoadjuvant, is associated with the presence of axillary node invasion at rescue surgery in locally advanced breast cancer. This might reflect the presence of intrinsically resistant clones before any form of therapy, which persist after it, and could be helpful both for prognosis and for the choice of individual treatment.

Endometrial carcinoma: assessment of myometrial invasion with plain and gadolinium-enhanced MR imaging.

The purpose of this study was to determine the accuracy of magnetic resonance imaging (MRI) for evaluating the depth of myometrial invasion, potential sources of pitfalls, and the usefulness of contrast‐enhanced series. Eighty‐five patients with a pathologic diagnosis of endometrial carcinoma underwent preoperative MRI (plain and contrast‐enhanced). Grade of myometrial invasion, presence of junctional zone (JZ), fibromyomas, and tumoral thickness were evaluated by two groups of radiologists blinded to pathologic results. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of MRI in determining the depth of myometrial invasion were evaluated. The sensitivity/specificity for plain MR was 64.1–64.1/93.5–100 for both observers. Assessing deep myometrial invasion, sensitivity, and NPV improved significantly (P = 0.002, P = 0.003 for both observers) when comparing plain and whole study series. Tumoral thickness (P = 0.16, P = 0.13, for the two observers) and presence or absence of JZ (P = 0.41, P = 0.14) did not influence myometrial invasion assessment. Gadolinium‐enhanced series improve the assessment of deep myometrial invasion in endometrial carcinoma. J. Magn. Reson. Imaging 2000;12:460–466.

Accumulation of uPA – PAI-1 complexes inside the tumour cells is associated with axillary nodal invasion in progesterone-receptor-positive early breast cancer

Both urokinase-like plasminogen activator (uPA) and its inhibitor plasminogen activator inhibitor (PAI-1), as well as uPA–PAI-1 complexes, have been identified as important prognostic factors in breast cancer. We have recently reported that the latter are identifiable inside breast cancer cells by means of immunohistochemistry. Using this technique, we have studied a series of 212 early (pT1) unifocal breast cancers and have correlated the expression of uPA–PAI-1 complexes, together with other clinical and biological features (histologic variety, histologic and nuclear grade, hormone receptors, Ki67 labelling index, c-erb-B2-, p53- and CD44std-expression) with or without the occurrence of axillary node invasion. In a logistic regression model, looking for associations with axillary metastasis, we found a statistically significant interaction between the presence of uPA–PAI-1 complexes and progesterone receptor positivity (P=0.04). A final model showed that the presence of uPA–PAI-1 complexes was a determinant factor for axillary metastasis among women carrying tumours expressing progesterone receptors. In these cases, the presence of uPA–PAI-1 complexes carried with it a nearly 14-fold risk of axillary node invasion (P=0.009). These results may indicate that small, hormone-receptor-positive breast cancers (with a theoretical good prognosis) may carry an elevated risk of nodal involvement if accumulation of uPA–PAI-1 complexes is shown inside their tumour cells by means of immunohistochemistry.

Cross-reactivity between Candida albicans and human ovarian carcinoma as revealed by monoclonal antibodies PA10F and C6.

Antibodies against Candida albicans antigenic determinants have been reported to cross-react with human tumour cells. We have found that two monoclonal antibodies, C6 and PA1OF, developed at our laboratory against C. albicans antigenic determinants, cross-react with human ovarian cancer on Western blots and immunohistochemistry. We have subsequently used one of them, PA10OF, to test by means of immunohistochemistry a series of 37 human ovarian carcinomas. Out of 37 tumours, 25 (67.6%) expressed the antigen recognized by PA1OF. The reactivity, however, was concentrated on the subgroup of particularly aggressive, invasive carcinomas in advanced stages of the disease (19 out of 24 positive), whereas the antigen was expressed significantly less (P=0.0007) in the subgroup of much less aggressive stage I tumours of low malignant potential, also called borderline carcinomas (2 out of 13 positive). This cross-reactivity between C. albicans and ovarian carcinoma seems to be attributable to a common antigenic determinant related to tumour aggressiveness.

Chemotherapy-induced emesis : management of early and delayed emesis in milder emetogenic regimens

Abstractu2002The objective of the present study was to examine the problem of the control of nausea and vomiting induced by non-cisplatin containing cyclophosphamide-based chemotherapy regimens in breast cancer patients. This was randomized, double-blind, parallel-group and placebo-controlled study comparing the efficacy of three antiemetic therapeutic regimens (ondansetron for 3 days, ondasetron plus metoclopramide, and ondansetron given in a single dose) in breast cancer patients receiving cyclophosphamide-based chemotherapy regimens on an outpatient basis. Both the primary and the secondary efficacy were measured. The primary efficacy variable was the number of emetic episodies (considering early and delayed emesis). The secondary efficacy variable measured was the quality of life. Two-by-two tables using the chi-square test and relative-risk concept were elaborated for statistical analysis. There was no difference between high-dose ondansetron and ondansetron plus metoclopramide among patients given CMF (cyclophosphamide, methotrexate, 5-fluorouracil). The single-dose ondansetron regimen showed the worst results. In patients given an FEC regimen (cyclophosphamide, epirubicin, 5-fluorouracil) the antiemetic efficacy was best for the high-dose ondansetron regimen, followed by the ondansetron plus metoclopramide regimen, and was worst for single-dose ondansetron administration. Despite the use of different antiemetic schedules, nausea and emesis are significant problems in patients receiving cyclophosphamide-based chemotherapy. Their adequate control should be the aim of any antiemetic approach.

82 P - CAl5.3: A breast cancer marker predicting location of metastases even before treatment

The aim of this study was to asses the ability of the antigenic tumor marker CA15.3 to identify, predict and exclude metastases in bone/viscera both in pretreatment determinations and dwing follow-up of breast cancer patients. Serum values of CA15.3 were measured in a prospective series of 164 patients prior to therapy and every 3 months during a minimum of 2 years-follow-up. In our series, 8 out of 14 (57.2%) and 51 out of 150 (34.0%) of patients with normal and elevated pretreatment values of CAl5.3 progressed (pxa0=xa00.085). Whereas 21 out of 51 patients (41.2%) who showed progression of the disease with pretreatment CA15.3 normal values developed bone metastases, 8 out of 8 (100%) patients with previous CA15.3 determinations superior to 40xa0U/ml and progression had bone metastases pxa0=xa00.0180) showing a predilection for this site of relapse in this group of patients. Considering the “follow-up determinations”, 27 patients out of 51 (52.9%) with CA15.3 pretreatment values inferior to 40xa0U/ml and progression showed CAl5.3 elevations during follow-up, CA15.3 correctly classified 64.4% of patients with progression (70.3% if only bone/viscera metastases are included) (sensitivity) and 94.3% without (specifity). Our results seem to indicate that patients with pretreatment CA15.3 high values are prone to develop bone metastases more frequently than other any other type of progression. There is the possibility that tumor heterogenety plays a role in this question. Cells expressing CAl 5.3 would possibly have more “bone-affinity”.

Elevated serum CA-125 levels in patients with ovarian fibrothecomas.

To DATE, CA-125 is the best tumour marker for ovarian adenocarcinoma [ 11. It has been found to be especially useful for monitoring the response to treatment of ovarian carcinoma patients with initially elevated CA-125 levels. Of them, 9&100% will have persistent ovarian cancer at second-look laparotomy if their serum CA- 125 fails to decline during treatment [2]. However, up to 50% will still have residual disease despite normal CA-125 levels, although in these cases the residual tumour is usually minimal [2]. Unfortunately, ovarian cancer is not the sole condition associated with elevated serum CA-125 levels. Other gynaecological cancers arising from the celomic epithelium (fallopian tube, endometrium, endocervix), non-gynaecological cancers (pancreatic, hepatic, breast, colon and lung), as well as pancreatitis, peritonitis, renal failure and liver cirrhosis, together with some benign gynaecological diseases (pelvic inflammatory disease, endometriosis) have all been associated with abnormal elevation of CA-125 [3]. This limits its usefulness as a screening tool for ovarian cancer in asymptomatic patients. Even in the presence of clinical signs suggestive of ovarian carcinoma, a high serum CA-125 is still not necessarily indicative of the disease. We have performed serial determinations of serum CA-125 levels in 6 consecutive patients with benign ovarian fibrothecomas diagnosed and treated at our hospital during the last 2 years. Initial serum CA-125 was invariably elevated in all patients before surgical treatment, in some of them extremely so (above 500 U/ml, which is the top limit at our laboratory). Serum levels had returned to normal in all patients after a maximum of 6 months following surgical excision (Fig. 1). The elevation of CA-125 in these patients seems to be associated with the proliferation of the rather inert fibrous stromal component of their tumours, since 1 patient with a pure ovarian thecoma diagnosed during the same period showed normal pretherapeutic CA-125 levels. The hypothesis is further supported by the findings of Walker et al. [4], who reported about 2 cases of ovarian cellular fibromas associated with an elevated CA-125. To corroborate it, we determined CA-125 serially in 5 patients subject to controlled ovarian hyperstimulation as part of an in V&J fertilisation program. Gonadotrophin hyperstimulation, as practised on these patients, often produces massively enlarged ovaries by acting mainly on the granulosa-cell population of them, and only secondarily affecting the inert stromal component. All 5 patients (data not shown) showed normal (below 30 U/ml) prestimulation and serial CA-125 levels throughout their stimulation cycles, also at the point of maximal stimulation, as determined by means of oestradiol plasma levels and ultrasound examination of follicle growth.

16 P Chemotherapy induced emesis. management of early and delayed emesis in milder emetogenic regimens

The objective of the study was to examine the problem of control of nausea and vomiting induced by non-cisplatin cyclophosphamide-based chemotherapic regimens in breast cancer patients. This was a randomized double-blind, parallel-group and placebo controlled study comparing the efficacy of four antiemetic therapeutic regimens (A:Ondansetron for 3 days; B:Ondasetron plus Metoclopramide; C:Granisetron given a single dosis and D:Ondansetron given in a single dosis) in breast cancer patients receiving Cyclophosphamide, Methotrexate and 5-Fluoracil (CMF) regimen (174 cycles) and Cyclophosphamide, 4-Epiadryamicin and 5-Fluoracil (FEC) regimen (132 cycles). Both, number of emetic episodies (early and delayed emesis) and quality of life were evaluated. In patients receiving CMF there were no differences between regimens A, B and C in controlling early emesis. The single dose Ondansetron regimen (D) showed the worst resuts (p=0.003). Delayed emesis was best controlled by the “3 days-regimens” (A and B). In patients administered a FEC treatment, the antiemetic efficacy was superior for the single Granisetron regimen (C) if early emesis was considered. Moreover, efficacy of single Granisetron dosis (C) was similar to the 3-days Ondansetron regimen (A) in controlling delayed emesis after FEC treatment. The single Ondansetron regimen showed again the worst results (p=0.007). Despite different antiemetic schedules, nausea and emesis are significant problems in patients receiving cyclophosphamide-based chemotherapy.