José Lastiri

Plasma metalloproteinase activity is enhanced in the euglobulin fraction of breast and lung cancer patients

Matrix metalloproteinases (MMP) have been implicated in tumor invasion and metastasis. We verified, by gelatin zymography, MMP activity in the euglobulin plasma fraction of 82 healthy controls, 66 patients with benign diseases and 149 patients with breast, lung, colon or brain cancer. The euglobulin fractions assayed showed 4 gelatinolytic bands of 62, 92, 120 and 200 kDa. The median (Md) value for 92 kDa‐MMP activity was significantly increased in breast (Md 1.34 arbitrary units [AU]/ml plasma, range 0.0–7.2) and lung cancer patients (Md 1.43 AU/ml, range 0.0–3.6) compared with the controls (Md 0.48 AU/ml, range 0.0–1.8). Patients with colon cancer or gliomas presented values of MMP‐9 similar to those of the healthy population. Multivariate analysis indicated that plasma MMP‐9 activity was not predicted by the known clinicopathological parameters such as age, stage, tumor size, number of positive lymph nodes, histologic grade, histologic type, nuclear grade or mitotic index. Lung cancer patients also presented high values of MMP‐9 (Md 1.43, range 0.0–3.6 [n = 26]), without association with tumor stage or histologic type. The levels of 92 kDa‐MMP activity in the plasma euglobulin fraction could be a potentially useful tumor marker in breast and lung cancer. Int. J. Cancer 89:389–394, 2000.

Mast cell phenotypes and microvessels in non–small cell lung cancer and its prognostic significance

The impact of interstitial inflammatory cells, such as mast cells, and angiogenesis on the prognosis of cancer patients has been reported in many solid tumors, although there is disagreement about their role. We undertook a retrospective study with tissue samples from 65 patients with stage I and II non-small cell lung cancer to assess the clinical pathologic role and prognostic significance of mast cells. Mast cell phenotypes were identified by immunohistochemistry for tryptase and chymase. In addition, we identified microvessels using the endothelial marker CD34. Mast cell and microvessel density was quantified by assessing immunopositive cells in the intratumoral and peritumoral zones of tumor specimens. Both mast cell and microvessel density was higher in the peritumoral zone than the intratumoral zone (P <or= .05). A positive correlation between mast cell (tryptase-chymase phenotype) and microvessel densities was observed in the intratumoral zone (P <or= .05), supporting the involvement of mast cells in the angiogenic process. Regarding survival, a subset of stage I patients had a worse prognosis at five years when low mast cell (tryptase-chymase phenotype) density was found in the peritumoral zone (median survival in months [range]: 27 [1-60] versus 46 [1-60]). Multivariate Cox analysis indicated that this parameter may be an independent prognostic factor (P <or= .05) useful for selecting candidates for earlier treatment.

Neural cell adhesion molecule in human serum. Increased levels in dementia of the Alzheimer type

Memory impairment is a process associated with alterations in neuronal plasticity, synapses formation, and stabilization. As the neural cell adhesion molecule (NCAM) plays a key role in synaptic bond stabilization, we analyzed the usefulness of soluble NCAM isoforms in the diagnosis of patients with dementia of the Alzheimer type (DAT). NCAM was measured in the sera of 70 control subjects and 43 DAT patients (with different severity of cognitive impairment, GDS), employing Western blot and densitometric quantification. LMW-NCAM bands (100-130 kDa) decreased significantly with age independently of sex. DAT patients presented values of LMW-NCAM and HMW-NCAM significantly higher than healthy controls of similar age (higher than 130 kDa). Only LMW-NCAM was associated with GDS. Our results suggest that NCAM could be involved in the pathogenesis of DAT disorder and that serum NCAM levels could be useful as differential diagnostic markers of the disease.

Prognostic value of Mdm2, p53 and p16 in patients with astrocytomas.

Surgical cure of gliomas infiltrating into the brain is practically impossible and their clinical course is primarily determined by the biological behavior of the tumor cell. The purpose of this study was to analyze retrospectively prognostic input of p53, Mouse double minute-2 (Mdm2) and p16 in 103 uniformly treated patients with astrocytic tumors. The expression of these molecules was measured by immunohistochemical procedure. Prognostic evaluation was performed with the multivariate proportional hazards model. The follow-up period lasted 19 (5–122) months for the survivors. We observed that 66% of gliomas showed mutated p53, while only 17% overexpressed Mdm2, the p53-regulatory molecule. Besides, almost 50% of gliomas lost p16 immunopositivity. Only p53 labeling showed a positive correlation with the grade of malignancy, according with the WHO classification. The association between mutated p53 and histological grade remained when prognostic variables were considered in a multivariate analysis. No association between p53 status and overall survival was found. On the other hand, Mdm2 overexpression and, unexpectedly, p16 immunopositivity were associated with a shorter survival in an univariate analysis. However, Cox-regression analysis showed that only Mdm2 in female patients was an independent prognostic factor, associated with shorter survival.In conclusion, our results suggest that Mdm2 could be a relevant marker in determining the evolution of glioma patients and could provide a more objective way to classify astrocytomas.

Nuevo Metodo de Regeneracion hepatica

Postoperative liver failure (PLF) is the most feared and serious complication after extensive liver resections. We present an innovative surgical technique for the treatment of a patient with colorectal cancer and initially unresectable liver metastases. After completing neoadjuvant chemotherapy, it was decided to perform simultaneous surgery. A left hemicolectomy and cleaning of the metastases in the left liver was performed. As the future liver remnant (FLR) was insufficient, it was decided to perform an in situ liver split and a right portal vein ligation. On the 6(th) day after the surgery a volumetric CT showed an increase greater than 40% of the FLR. The right hepatectomy was completed and the patient was discharged on the 11(th) day after surgery. The technique induced a rapid growth of the FLR, exceeding that reported using portal occlusion. If these findings are corroborated in future studies, this revolutionary technique could enable surgery to be performed in two stages on patients with initially unresectable liver disease during the same hospital admission and without PLF.

Colorectal Cancer Staging: Reappraisal of N/PN Classification

PURPOSE: Current American Joint Committee on Cancer and the Union Internationale Contre le Cancer TNM classification disregards location of positive nodes, discontinuing N3 category, which constitutes a major modification to 1987 version. This study was designed to assess the impact of the recategorization of former N3 cases and the reliability of the current N1-N2 subcategorization of Stage III patients. METHODS: Prospectively collected data from 1,391 patients (55.8 percent males; median age, 64 (range, 21–97) years), operated on with curative intent between 1980 and 1999, were analyzed. The median follow-up was 60 (interquartile range, 27–97) months with 129 cases lost to follow-up. RESULTS: Of positive node cases, 25.3 percent were former N3. Among them, 30.5 percent migrated to the N1 group and 69.5 percent to the N2 group. The proportions of former N3 cases in N1 and N2 groups were 12.5 percent and 46.1 percent, respectively (P < 0.001). Node-positive patients had an actuarial five-year survival rate of 56.7 percent (95 percent confidence interval, 53–59), with a significant difference between N1/N2 categories (63.6 vs. 44.1 percent, respectively; P < 0.001). Although apical node involvement and more than three positive nodes were associated with poorer outcomes in univariate analysis, only the number of positive nodes had independent association (hazard ratio, 1.6 (range, 1.2–2.2); P < 0.001). Integration of former N3 cases did not modify outcomes. CONCLUSIONS: The recategorization of former N3 involved a high proportion of positive node cases. Current N1/N2 categories clearly defined different outcomes and were not modified by the integration of former N3.

The neural cell adhesion molecule is involved in the metastatic capacity in a murine model of lung cancer.

Neural cell adhesion molecule (NCAM) is involved in cell growth, migration, and differentiation. Its expression and/or polysialylation appear to be deregulated in many different cancer types. We employed the lung tumor cell line LP07, syngeneic in BALB/c mice to investigate the role of NCAM in malignant progression. LP07 cells express the three main NCAM isoforms, all of them polysialylated. This cells line, pretreated with an anti‐NCAM antibody and inoculated intravenously (i.v.) into syngeneic mice, developed less and smaller lung metastases. In vitro studies showed that NCAM bound antibody inhibited cell growth, mainly due to an increase in apoptosis, associated with a decrease of cyclin D1 and enhanced expression of active caspase 3 and caspase 9. Anti‐NCAM‐treated LP07 cells showed impairment in their ability to migrate and adhere to several extracellular matrix components. Secreted uPA activity was also reduced. NCAM‐140 knocked‐down by siRNA in LP07 cells pretreated or not with anti‐NCAM showed an impaired metastasizing ability upon i.v. inoculation into mice. These results suggest that anti‐NCAM treatment could be mimicking homophilic trans‐interactions and NCAM‐140 knocked‐down impairs heterophilic interactions, both leading to inhibition of metastatic dissemination. The involvement of NCAM in lung tumor progression was confirmed in human NSCLC tumors. Sixty percent of the cases expressed NCAM at tumor cell level. A multivariate analysis indicated that NCAM expression was associated with a shorter overall survival in this homogeneous series of Stages I and II NSCLC patients. NCAM may be able to modulate mechanisms involved in lung carcinoma progression and represents an attractive target to control metastatic progression. Mol. Carcinog.

Association between mast cells of different phenotypes and angiogenesis in colorectal cancer

It is known that mast cells proliferate in solid tumours and increase tumour angiogenesis. Nevertheless, there is no consensus regarding their role in colorectal cancer (CRC). In this study, we aimed to clarify the relationship of mast cells positive for tryptase (MCts) and tryptase-chymase (MCtcs) with microvessel density (MVD) in the intratumoral zone and the invasive edge of 80 CRC patient tumours. We evaluated these parameters and associated their expression with clinicopathological parameters, including survival rate. Tumour sections from each patient were immunostained for tryptase to evaluate MCts, chymase to evaluate MCtcs, and CD34 to evaluate microvessel counts under x100 microscopy. The number of MCs of both phenotypes and the MVD counts were higher in the invasive edge than in the intratumoral zone (p<0.001). MCt numbers were higher than those of MCtcs in all Astler-Coller stages in both regions. A positive correlation between MVD and MCts or MCtcs was observed (Pearsons test p<0.001). Neither the number of MCs nor MVD was associated with overall survival (log rank test). However, only 8.3% of patients with low numbers of MCtcs in the invasive edge succumbed to the disease, compared to 32% with high numbers of MCtcs. Our results indicate that angiogenesis and MC hyperplasia are events which appear early during CRC development. The correlation of MC phenotypes with MVD is in agreement with the role attributed to MCs, that of angiogenesis enhancement. Collectively, these findings suggest that screening during the early malignization of CRC can provide valuable clinical information.

Tumor size as prognostic factor in osteosarcomas

9061 Background: Multidisciplinary treatment of osteosarcoma has achieved overall survival greater than 70%. Several prognostic factors have been described. In our country it is frequent to diagnose voluminous tumors. Greater tumors usually have worst outcome because of greater chance of tumor resistance. In order to assess the relation between tumor size and prognosis in high-grade osteosarcoma, we performed the following analysis. METHODS Retrospectively, tumor volume was determined pre and post chemotherapy in 40 patients with histological diagnosis of high-grade osteosarcoma stage IIb. All of them received neo-adjuvant chemotherapy (ifosfamide + doxorrubicin + high dose methotrexate). Tumor size was determined in plain radiography (RX) (2 dimensions) and magnetic resonance imaging (MRI) (3 dimensions), according to RECIST criteria. Tumor calcifications, pathological fracture and skip metastasis were evaluated. RESULTS 22 men/18 women. Median age 17 years (range 11-47). Localizations: femur (25), tibia (9), fibula (3), humerus (2) and pelvis (1). Six patients presented pathological fracture and only one skip metastasis. 14 patients had systemic progression. Median survival was 96 months (IC 95%: 80-109) and median disease free survival was 84 months (IC 95%: 69-100). Tumor volume pre chemotherapy greater than >1500 cm3 (MRI) was the variable most predictor of mortality (p=0.02). Median survival was 48 months (IC 95%: 27-70) in tumors greater than 1500 cm3 versus 104 months (IC 95%: 91-118) in those tumors smaller than 1500cm3 (p=0.001). Intensification of radiological calcification, pathological fracture and changes in size pre and post chemotherapy did not have prognostic value. CONCLUSIONS tumor measurement at diagnosis can distinguish high-risk populations. MRI is superior to RX to estimate prognostic size value. MRI does not sub estimate soft parts of the tumor and allows delimiting better tumor margins. No significant financial relationships to disclose.