A 50-gene signature is a novel scoring system for tumor-infiltrating immune cells with strong correlation with clinical outcome of stage I/II non-small cell lung cancer.

PURPOSE: To identify a novel system for scoring intratumoral immune response that can improve prognosis and therapy decisions in early stage non-small cell lung cancer (NSCLC). METHODS/PATIENTS: Eighty-four completely resected stage I/II NSCLC without adjuvant therapy were classified by expression profiling using whole genome microarrays. An external cohort of 162 tumors was used to validate the results. Immune cells present in tumor microenvironment were evaluated semiquantitatively by CD20, CD79, CD3, CD8, CD4 and CD57 immunostaining. Univariate and multivariate analyses of variables associated with recurrence-free survival were performed. RESULTS: Initial molecular classification identified three clusters, one with significantly better RFS. A reduced two-subgroup classification and a 50-gene predictor were built and validated in an external dataset: high and low risk of recurrence patients (HR = 3.44; p = 0.001). Analysis of the predictor´s genes showed that the vast majority were related to a B/plasma cell immune response overexpressed in the low-risk subgroup. The predictor includes genes coding for unique B lineage-specific genes, functional elements or other genes that, although non-restricted to this lineage, have strong influence on B-cell homeostasis. Immunostains confirmed increased B-cells in the low-risk subgroup. Gene signature (p < 0.0001) and CD20 (p < 0.05) were predictors for RFS, while CD79 and K-RAS mutations showed a tendency. CONCLUSIONS: Favorable prognosis in completely resected NSCLC is determined by a B-cell-mediated immune response. It can be differently scored by a 50-gene expression profile or by CD20 immunostaining. That prognosis information not reflected by traditional classifications may become a new tool for determining individualized adjuvant therapies.

Celecoxib in a 12-year-old boy with familial adenomatous polyposis.

Familial Adenomatous Polyposis (FAP) is an autosomal dominant disorder characterized by colonic polyps in early adult life. Children with this disease are at risk for colonic cancer, so prophylactic colectomy is the standard treatment to prevent this complication. Chemoprevention experience with NSAIDs in children is exceptional. This case report describes our experience with Celecoxib, a COX-2 inhibitor, in a 12-year-old boy.

Molecular analysis of colorectal cancer tumors from patients with mismatch repair proficient hereditary nonpolyposis colorectal cancer suggests novel carcinogenic pathways.

PURPOSE: A subset of colorectal cancers (CRC) arises in families that, despite fulfilling clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC), do not show evidence of a mismatch repair (MMR) deficiency. The main objective of this study was to characterize these tumors at the molecular level. EXPERIMENTAL DESIGN: After comprehensive germ line mutation scanning, microsatellite analysis, and MMR protein expressions, we selected a well-defined cohort of 57 colorectal tumors with no evidence of MMR defects. In this group of tumors, we analyzed KRAS, BRAF, and APC somatic mutations, as well as methylguanine methyltransferase (MGMT) and beta-catenin expression. We correlated these alterations with clinicopathologic data and explored the relationship between KRAS G > A transitions and lack of MGMT expression. RESULTS: The mutation profile at the RAS/RAF/MAPK pathway mimics sporadic microsatellite-stable CRCs. We found an average age of diagnosis 10 years older in KRAS-mutated patients (P = 0.001). In addition, we show that KRAS G > A transitions are actively selected by tumors, regardless of MGMT status. Similarities with HNPCC high-microsatellite instability tumors are observed when APC data are analyzed. The APC mutation rate was low and small insertions/deletions accounted for 70% of the alterations. In addition, we found a low frequency of beta-catenin nuclear staining. Finally, we did not find evidence of tumors arising in individuals from the same family sharing molecular features. CONCLUSIONS: We show evidence that CRC tumors arising in HNPCC families without MMR alterations have distinctive molecular features. Overall, our work shows that systematic analysis of somatic alterations in a well-defined subset of CRCs is a good approach to provide new insights into the mechanisms of colorectal carcinogenesis.

P53 overexpression predicts endometrial carcinoma recurrence better than HER-2/neu overexpression.

OBJECTIVE: to investigate the prognostic value of p53 and HER-2/neu overexpression in endometrial cancer. STUDY DESIGN: p53 and HER-2/neu immunostaining was performed in 114 paraffin-embedded specimens of endometrial cancer diagnosed and treated between 1990 and 1997. Nuclear p53 and membrane HER-2/neu immunostaining were used. RESULTS: p53 and HER-2/neu overexpression was observed in 17 cases (14.9%) and in 19 cases (16.7%), respectively. In univariate analysis p53 (P<0.001) and HER-2/neu (P=0.018) overexpression had a positive correlation with a high risk of recurrence. In multivariate analysis, age (P<0.001), FIGO stage (P<0.001), differentiation (P=0.013), non-endometrioid subtypes (P<0.001) and p53 overexpression (P<0.001), but not HER-2/neu overexpression, were independent prognostic indicators of recurrence. Simultaneous p53 and HER-2/neu overexpression made worse the prognostic (P<0.001). CONCLUSIONS: p53 overexpression was an independent predictor of recurrent disease in endometrial cancer. HER-2/neu overexpression had a more limited effect but enhance the effect of p53.

Cyclophosphamide induces the development of early myeloid cells suppressing tumor cell growth by a nitric oxide-dependent mechanism.

Adoptive immunotherapy with cyclophosphamide (Cy) increases the host resistance against tumor growth. The precise mechanism(s) by which this therapy enhances tumor suppression is unclear. Cy induces the development of early myeloid cells that may be strongly antiproliferative through NO production. These cells are similar to the natural suppressor cells found in normal bone marrow with a potential antitumor effect. Here we have addressed whether the development of NO-producing cells may be involved in this tumor resistance in Cy-treated mice. The results show a synergism between Cy treatment and tumor-specific lymphocytes transferred systemically (i.v.) or locally (Winn's assay) that results in a strong tumor suppression. Inhibition of NO production by N(G)-monomethyl-L-arginine at the site of tumor inoculation results in a loss of the protection achieved by the combined therapy. Cy-treated mice develop splenic early myeloid (CD11b, Gr-1, CD31 (ER-MP12), ER-MP20, ER-MP54) cells producing large amounts of NO upon T cell-derived signals (IFN-gamma plus CD40 ligation) able to inhibit tumor cell growth in vitro. Early myeloid cells (ER-MP54(+)) and cells expressing inducible NO synthase are increased at the site of tumor challenge in mice treated with the combined therapy, but not in those treated with Cy or immune cell transfer alone. Thus, Cy induces the expansion of early myeloid cells, inhibiting tumor cell growth by a mechanism involving NO. Both the recruitment and the activation of these myeloid cells at the site of tumor challenge appear to be dependent on the presence of tumor-specific lymphocytes.

[The cost effectiveness of echography biopsy]. / Rentabilidad de la ecopsia.

Ecopsy is a postmortem technique which, by means of echography-guided puncture and/or aspiration obtains material for histological analysis. This study compared cost and time employed in 100 ecopsies and 100 classic necropsies and confirmed that cost of materials in ecopsy is 65% lower than that in necropsy. Physicians, necropsy technicians, laboratory technicians and secretary team personnel spent 33%, 54%, 19% and 32% less time than in necropsy. The clinical report of ecopsy was finished within nine days even with the brain study included.

Prognostic significance of p53 gene mutations in squamous cell carcinoma of the lung.

The aim of the present study was to analyze the prevalence and clinical importance of p53 gene mutations in surgically treated squamous cell lung carcinoma. Sixty patients were included. Fifty-one patients in stages I to IIIa were submitted to radical resection. Twenty-five samples tested positive for the p53 immunohistochemistry assay, and were analyzed for p53 gene mutations. Eleven mutations were found. Patients harboring p53 gene mutations suffered a higher incidence of recurrence and a higher mortality rate. Disease-free interval and overall survival were shorter for patients with mutated p53 gene (p=0.03 and p=0.005, respectively).

Prognostic value of flow cytometric DNA analysis in non-small-cell lung cancer: rationale of sequential processing of frozen and paraffin-embedded tissue.

The objective of this study was to determine the prognostic information provided by flow cytometric DNA analysis in non-small-cell lung cancer. Lung samples of 132 consecutive patients submitted to surgery were prospectively processed. When no aneuploid populations were detected in fresh frozen samples, the process continued as a second step in paraffin-embedded tissue, consuming all the tumor available. The influence of ploidy on the postoperative outcome was studied by both a univariate and a multivariate analysis. Aneuploidy was found in 81 patients (61.4%). Fourteen patients showed no aneuploidy in fresh frozen samples; and only after further analysis in paraffin-embedded tissue was abnormal DNA detected. Overall, the 36-month survival was 69% for the diploid group and 24% for the aneuploid group (p = 0.0006). Including subjects submitted to complete tumor removal (stages I, II, and IIIA) in a multivariate analysis adjusted for TNM stage and histologic type, bearers of aneuploid tumors exhibited a higher risk of relapse (hazard ratio 2.65; CI 95% 1.5-4.66;p = 0.004) or death (hazard ratio 2.17; CI 95% 1.08-4.39;p = 0.032) than patients with diploid tumors. DNA ploidy resulted an independent prognostic factor of survival and tumor relapse in completely resected non-small-cell lung cancer. Sequential analysis of fresh and paraffin-embedded samples can help avoid the bias due to intratumoral DNA content heterogeneity. DNA ploidy could be an useful parameter in any future multifactorial analysis of outcome in such tumors.

[Recurrent ascites in peritoneal mesothelioma. Its diagnostic and therapeutic management]. / Ascitis recidivante en el mesotelioma peritoneal. Manejo diagnóstico y terapéutico.

Peritoneal mesothelioma is an uncommon neoplasm often related to previous asbestos exposure. It is necessary to exclude other secondary peritoneal neoplasm. The application of immunohistochemical analysis in the biopsy sample is important for establishing an accurate diagnosis. We report the case of a peritoneal mesothelioma that started as a haemorrhagic ascites. After laparotomy, the initial diagnosis was peritoneal carcinomatosis from adenocarcinoma of unknown origin. The diagnosis was obtained by using immunohistochemical analysis: vimentin and keratine antibodies were positive and leu M1, antibodies were negative. The interest of our case resides in the difficulty for obtaining the diagnosis and the complicate management of refractory ascites. Our patient required intraperitoneal 5-fluorouracil for controlling the ascites.

[Ascites due to cerebrospinal fluid accumulation]. / Ascitis por acúmulo de líquido cefalorraquídeo.

We present the clinical case of a 17-year-old male who developed an ascitic syndrome 11 years after the establishment of a ventricular-peritoneal shunt as the consequence of a malformation hydrocephalus. We discuss the pathogenic mechanism and present histopathological evidence that justifies the accumulation of spinal fluid in the peritoneal cavity. We insist on the convenience of keeping in mind this clinical situation in order to avoid unnecessary complementary exams and we analyze the most adequate therapeutic schedule.