Effect of ß-catenin alterations in the prognosis of patients with sporadic colorectal cancer.

CONTEXT: Wnt pathway activation represents a critical step in the etiology of most of colorectal cancer (CRC) and it is commonly due to mutations in the APC gene, which originates the loss of ß-catenin regulatory function. It has been suggested that APC inactivation or ß-catenin alteration have similar effects in tumor progression in CRC tumorigenesis. AIMS: The aim of this study was to analyze the frequency of ß-catenin gene mutation in patients with sporadic CRC and to determine its effect in prognosis. MATERIALS AND METHODS: This was a prospective cohort study, which included 345 patients with sporadic CRC. ß-Catenin gene mutations in exon 3 were detected by single strand conformation polymorphism (SSCP). Exon 3 deletion was studied by identifying differences in fragment length of specific amplification products. All the altered samples were confirmed by direct sequencing. RESULTS: In our population, point mutations were detected in 1.8% of the samples and 4.9% of the samples showed deletion. We observed association between exon 3 mutations and increased levels of Carcinoenbryonic Antigen (CEA). In these patients, clinically relevant improvement in overall survival was also observed. CONCLUSION: Frequency of point mutations in exon 3 ß-catenin gene is low in our population. It would be interesting to increase the population size to test the clinically relevant influence in the prognosis found, and to test the relation of these events with Microsatellite Instabillity (MSI) pathway. If these findings were confirmed, ß-catenin determination would help in the selection of patients with different prognosis.

A9 region in EPHB2 mutation is frequent in tumors with microsatellite instability. Analysis of prognosis.

AIM: The aim of the present study was to determine the relation of EPH tyrosine kinase receptor B2 (EPHB2) A9 region mutation and microsatellite instability (MSI); and to analyze their influence in prognosis of patients with sporadic colorectal cancer (CRC). PATIENTS AND METHODS: A total of 481 patients with CRC were examined. MSI (NCI criteria) and EPHB2 were analyzed using PCR and fragment analysis software. RESULTS: EPHB2 mutation was detected in 3.1% of patients. Mutation of EPHB2 was associated with location and with MSI status. We considered low instability (L-MSI) when only one marker showed instability, high instability (H-MSI) when two or more markers were positive and microsatelllite stable (MSS) when no instability was detected. The stratified analysis of overall survival (OS) and disease-free survival (DFS) in MSI according to EPHB2 status revealed no statistically significant differences. However, the risk of recurrence of H-MSI tumors with EPHB2 mutation carriers was 3.6-times higher than in non-mutation carriers. CONCLUSION: The frequency of EPHB2 mutation is higher in patients with H-MSI than MSS tumors. Promising results were found regarding the prognostic influence of EPHB2 in H-MSI.

Serum level of fibroblast growth factor 23 in maintenance renal transplant patients.

BACKGROUND: The discovery of fibroblast growth factor 23 (FGF23) provides a new conceptual framework that improves our understanding of the pathogenesis of post-transplant bone disease. Excess FGF23 is produced in the early post-transplant period; levels return to normal in the months following transplant. However, few manuscripts discuss FGF23 levels in stable long-term renal transplant recipients. METHODS: We performed a cross-sectional observational study of 279 maintenance kidney recipients with chronic kidney disease (CKD) Stages 1-4 and stable allograft function who had received their transplant at least 12 months previously. We calculated the estimated GFR (eGFR) using the MDRD4 equation. RESULTS: FGF23, parathyroid hormone (PTH) and phosphorus values were higher in more advanced stages, while the serum calcitriol levels and the phosphate reabsorption rate were lower. A significant inverse correlation was found between eGFR and FGF23 (r = -0.487; P < 0.001), PTH (r = -0.444; P < 0.001), serum phosphate levels (r = -0.315; P < 0.001) and fractional excretion of magnesium (r = -0.503; P < 0.001). Multivariable analysis showed that increased time on corticosteroids (P < 0.001), PTH (P < 0.001), serum phosphate (P = 0.003), decreased serum calcitriol (P = 0.049) and estimated glomerular filtration (P = 0.003) rate were associated with high FGF23 levels. In contrast with pre-transplant patients and first year post-transplant patients, higher FGF23 values were not correlated with increased phosphate excretion. An elevated phosphate reabsorption rate was associated with decreased PTH (P < 0.001) and calciuria (P = 0.028) and increased serum calcitriol (P = 0.009), plasma bicarbonate (P = 0.024) and estimated glomerular filtration (P = 0.003). CONCLUSIONS: Serum FGF23 concentrations remain increased in long-term kidney graft recipients, even in the early stages of CKD. It remains to be seen whether measures aimed at reducing serum levels of PTH and phosphate and/or corticosteroid doses might help to lower serum FGF23 and whether this will improve kidney recipient outcomes.

Detección rápida de metástasis en ganglio centinela y comparaciónde diferentes técnicas en carcinomas de mama de bajo grado / Rapid detection of sentinel lymph node metastases in different techniques and comparison in low-grade breast carcinomas

Introduction: The role of sentinel node biopsy has revolutionized breast cancer treatment. This determinationreduces the mobility of a complete axillary lymphadenectomy. The aim of our study is to analyze the value of sentinel node in low-grade histological breast tumors, studied with hematoxylin and eosin techniques, mmunohistochemistry,and molecular chain reaction in real-time quantitative polymerase (RT-PCR). Materials and methods: In a pilot study we studied a total of 21 patients with histological diagnosis of mucinous carcinoma, adenoid cystic carcinoma, and medullar carcinoma that underwent the sentinel node technique. Once the lymph node was removed, it was sent to pathology, where it was fragmented and evaluated, using between 25% and 50% of the lymph node for molecular biology laboratory studies. Results: The sentinel nodes studied were 32, corresponding to the 21 patients. Of the 32 lymph nodes analyzed, 29 (90.6%) were negative on histopathological examination and the molecular identification, 2 (6.2%) were positive in both techniques and 1 (3.125%) lymph node was positive with quantitative RT-PCR and negative in histology (H&E), which – subsequently by immunohistochemistry (IHC) – was diagnosed as isolated tumor cells (ITC). Conclusion: When comparing the techniques of hematoxylin and eosin, immunohistochemistry, and molecularRT-PCR technique, we found greater sensitivity of molecular techniques; this can reduce the false negative andimprove diagnosis of sentinel node metastases. Patients with low histological grade carcinomas have high survivalrates, less aggressive tumor behavior, and reduced lymph node at diagnosis.

Circulating tumor cells in solid tumor in metastatic and localized stages.

UNLABELLED: The aim of this study was the detection of circulating tumor cells (CTC) in three tumor types of epithelial origin. PATIENTS AND METHODS: Four hundred and thirty-eight patients with breast cancer (56.2% localized and 43.8% metastatic), 195 with colorectal tumors (84.1% localized and 15.9% metastatic) and 50 with prostate cancer (52% localized and 48% metastatic) took part in this study. CTC quantification was performed using the CellSpotter Analyzer (Veridex LLC). RESULTS: 31.5% of patients with cancer had > or =2 CTCs/7.5 mL but none of the healthy volunteers were above this level (p<0.001). Among patients with metastatic disease, 62.3% of them had > or =2 CTCs/7.5 mL but only 14.0% of those with localized disease were above this level (p<0.001). The presence of CTCs were correlated to stage in the three studied tumor types and no differences in the number of cells were found between them. CONCLUSION: The presence of more than 2 CTCs/7.5 ml is a frequent event in metastatic cases. In particular, patients with localized disease who have more than 2 CTCs/7.5 ml should be carefully studied to determine the possible prognostic and predictive value of this finding.

Circulating tumour cells in locally advanced breast cancer.

MATERIAL AND METHODS: A prospective study was conducted to determine the value of changes in circulating tumour cell (CTC) levels prior to and after the first cycle of neoadjuvant treatment in early prediction of pathologic response in locally advanced breast cancer (LABC). Two blood samples were obtained from 72 eligible LABC patients to isolate and enumerate CTCs before neoadjuvant chemotherapy started on day 1, and on day 21, immediately before second cycle administration. RESULTS: Sixty patients (83.3%) had <1 CTC in the first sample and response rates in this cohort were pathologic complete response (PCR) in 2 patients (5%), partial response (PR) in 35 (87.5%), stable disease (SD) in 2 (5%) and progressive disease (PD) in 1 (2.5%). Twelve patients (16.7%) had >2 CTCs in the first sample; these patients were more likely to have triple negative tumours. All 12 had fewer CTCs in the second sample. Response rates in this second cohort of 12 patients were PCR in 4 (34%), PR in 6 (50%), SD in 1 (8%) and PD in 1 (8%). PCR rate was markedly better in this second cohort (p<0.0042; OR 14.5, 95% CI 2.3-92). DISCUSSION: This study suggests that the presence of CTCs prior to neoadjuvant therapy might be a predictor of response to this therapy.

Inactivation of p16 by CpG hypermethylation in renal cell carcinoma.

OBJECTIVE: Renal carcinoma develops as a consequence of the accumulation of several genetic aberrations. Alterations in the p16 gene have been described in many tumors. Methylation of its promoter in CpG islands is the most common mechanism of inactivation of this gene. The aim of this study was to establish whether p16 gene methylation leads to a loss of the encoded protein in 57 patients with renal carcinoma, and if this aberration has any value in predicting disease progression in these patients. METHODS: Gene promoter methylation was determined by deoxyribonucleic acid treated with sodium bisulfite to subsequently amplify methylated and unmethylated regions rich in CpG islands. The p16 protein product was detected for immunohistochemical examination. RESULTS: Hypermethylation of the p16 gene was detected in 22.9% of the patients, none of whom had the protein product. A lack of p16 protein was confirmed in 52.9% of the tumors, indicating another genetic alteration or posttranscriptional modifications preventing the codification of this protein. Through multivariate analysis of overall survival, gene methylation was found to have independent prognostic value: the absence of alteration confers an undefined risk of death. CONCLUSIONS: Of the molecular modifications described for renal carcinoma, aberrations in the p16 gene are frequent. In these patients, methylation of the p16 gene promoter seems to afford a protective effect against the risk of death.

[Detection and quantification of circulating tumor cells in peripheral blood in patients with colon cancer]. / Detección y cuantificación de células tumorales en sangre periférica en pacientes con cáncer de colon.

BACKGROUND AND OBJECTIVE: The aim of this study is the detection and quantification of circulating tumor cells (CTC) in patients diagnosed with colon cancer and to establish whether they are related to the main clinicopathologic variables for this type of carcinoma. PATIENTS AND METHOD: Twenty-five colon cancer patients and 30 healthy volunteers were analysed. The quantification was performed using the CellSpotter Analyzer (Veridex LLC), that allows immunomagnetic isolation and immunospecific labelling of the cells for their enumeration. RESULTS: 72% of the colon cancer patients showed CTC and the mean number of cells found was 5 CTC/7.5 ml of peripheral blood. 52% of the samples contained 2 or more cells. Considering 2 cells as the cut-off point, a significant relationship with lactate dehydrogenase was found. CONCLUSIONS: This new technology which allows isolation and quantification of CTC in peripheral blood has proven to be valid for the detection of epithelial cells in colon cancer patients in every tumor stage. The results shown in this work confirm that cytokeratin 8, 18 and 19 are detected in CTC in this tumor type and will allow us to develop a protocol for the study of the relationship of quantification of theses cells and the clinical parameters involved in colon cancer.

Detección y cuantificación de células tumorales en sangre periférica en pacientes con cáncer de colon / Detection and quantification of circulating tumor cells in peripheral blood in patients with colon cancer

Fundamento y objetivo: El objetivo de este estudio es cuantificar las células tumorales circulantes (CTC) en pacientes diagnosticados de cáncer de colon y determinar su relación con las variables clinicopatológicas de interés en este tipo de carcinomas. Pacientes y método: La población del estudio está formada por 25 pacientes con cáncer de colon y 30 personas voluntarias sanas. El análisis de las CTC se realiza con el CellSpotter Analyzer (Veridex LLC), que aísla y determina las células mediante técnicas inmunomagnéticas. Resultados: El 72% de los pacientes con cáncer de colon estudiados presentan CTC, con una media de 5 células/7,5 ml de sangre periférica. El 52% presenta 2 o más de 2 CTC. Con este punto de corte, se observa relación de la determinación de CTC con la lactatodeshidrogenasa. Conclusiones: Este nuevo sistema de aislamiento y cuantificación de CTC en sangre periférica permite la detección de células epiteliales en pacientes con adenocarcinoma de colon de todos los estadios tumorales. Los resultados de este estudio confirman que con las citoqueratinas 8, 18 y 19 se detectan las CTC en este tipo de tumores y nos permite establecer un protocolo para estudiar la relación de la cuantificación de estas células con los parámetros clínicos del cáncer de colon

Background and objective: The aim of this study is the detection and quantification of circulating tumor cells (CTC) in patients diagnosed with colon cancer and to establish whether they are related to the main clinicopathologic variables for this type of carcinoma. Patients and method: Twenty-five colon cancer patients and 30 healthy volunteers were analysed. The quantification was performed using the CellSpotter Analyzer (Veridex LLC), that allows immunomagnetic isolation and immunospecific labelling of the cells for their enumeration. Results: 72% of the colon cancer patients showed CTC and the mean number of cells found was 5 CTC/7.5 ml of peripheral blood. 52% of the samples contained 2 or more cells. Considering 2 cells as the cut-off point, a significant relationship with lactate dehydrogenase was found. Conclusions: This new technology which allows isolation and quantification of CTC in peripheral blood has proven to be valid for the detection of epithelial cells in colon cancer patients in every tumor stage. The results shown in this work confirm that cytokeratin 8, 18 and 19 are detected in CTC in this tumor type and will allow us to develop a protocol for the study of the relationship of quantification of theses cells and the clinical parameters involved in colon cancer

Circulating tumoral cells lack circadian-rhythm in hospitalized metastasic breast cancer patients.

BACKGROUND: The relationship between breast cancer and circadian rhythm variation has been extensively studied. Increased breast tumorigenesis has been reported in melatonin-suppressed experimental models and in observational studies. OBJECTIVES: Circulating Tumor Cells (CTC) circadian- rhythm may optimize the timing of therapies. This is a prospective experimental study to ascertain the day-time and night-time CTC levels in hospitalized metastasic breast cancer (MBC) patients. MATERIAL AND METHODS: CTC are isolated and enumerated from a 08:00 AM and 08:00 PM blood collections. 23 MBC and 23 healthy volunteers entered the study. 69 samples were collected (23 samples at 08:00 AM and 23 samples at 08:00 PM from MBC; 23 samples from healthy volunteers). Results from two patients were rejected due to sample processing errors. No CTC were isolated from healthy-volunteers. RESULTS AND DISCUSSION: No-differences between daytime and night-time CTC were observed. Therefore, we could not ascertain CTC circadian-rhythm in hospitalized metastasic breast cancer patients.

[Microsatellite instability predicts better outcome in colorectal cancer patients]. / La inestabilidad de microsatélites predice un mejor pronóstico en los pacientes intervenidos de carcinoma colorrectal.

BACKGROUND AND OBJECTIVE: Two different pathways for the development of tumor have been described in colorectal carcinoma: the chromosomic instability, raised by suppressor genes and proto-oncogene alterations, and the microsatellite instability (MSI), caused by alterations in DNA repairing genes. PATIENTS AND METHOD: The frequency and the clinical meaning of the microsatellites instability pathway were determined in a consecutive prospective cohort of 106 patients who underwent surgical resection of colorectal carcinoma by a single surgeon. Microsatellite instability determination was established according to the criteria proposed by the National Cancer Institute in 1998. RESULTS: 9.4% of patients had a high instability and it was low in 11.3%; both groups displayed different clinico-pathological characteristics (age, sex, tumor site and histologic type). In the multivariant analysis of overall survival and disease free survival, high instability exhibited prognostic value independent of the rest of variables evaluated (p < 0.0001). CONCLUSIONS: The genetic alterations giving rise to microsatellite instability lead to a better prognosis in patients with colorectal cancer.

La inestabilidad de microsatélites predice un mejor pronóstico en los pacientes intervenidos de carcinoma colorrectal / Microsatellite instability predicts better outcome in colorectal cancer patients

FUNDAMENTO Y OBJETIVO: En el carcinoma colorrectal se describen 2 vías genéticas diferentes implicadas en la génesis del tumor: la inestabilidad cromosómica, debida a la alteración de genes supresores o protooncogenes, y la inestabilidad de microsatélites, originada por alteraciones en los genes reparadores del ADN. PACIENTES Y MÉTODO: En este estudio se determina la frecuencia y el significado clínico de la vía de la inestabilidad de microsatélites en una cohorte prospectiva consecutiva de 106 pacientes intervenidos por carcinoma colorrectal por un mismo cirujano. Para la determinación de la inestabilidad de microsatélites se han seguido los criterios propuestos por el National Cancer Instituteen 1998. RESULTADOS: El 9,4% de los pacientes muestra una alta inestabilidad y el 11,3% una inestabilidad baja. Ambos grupos presentan diferentes características clínico patológicas (edad, sexo, localización del tumor y tipo histológico). En el análisis multivariante de la supervivencia global y de la supervivencia libre de enfermedad, la alta inestabilidad presenta un valor pronóstico independiente del resto de las variables clínico patológicas analizadas (p < 0,0001). CONCLUSIONES: La alteración genética que supone la alta inestabilidad de microsatélites confierea los pacientes con cáncer colorrectal un mejor pronóstico

BACKGROUND AND OBJECTIVE: Two different pathways for the development of tumor have been described in colorectal carcinoma: the chromosomic instability, raised by suppressor genes and protooncogene alterations, and the microsatellite instability (MSI), caused by alterations in DNA repairing genes. PATIENTS AND METHOD: The frequency and the clinical meaning of the Microsatellites instability pathway were determined in a consecutive prospective cohort of 106 patients who underwent surgical resection of colorectal carcinoma by a single surgeon. Microsatellite instability determination was established according to the criteria proposed by the National Cancer Institute in1998. RESULTS: 9.4% of patients had a high instability and it was low in 11.3%; both groups displayed different clinico-pathological characteristics (age, sex, tumor site and histologic type). In the multivariant analysis of overall survival and disease free survival, high instability exhibited prognostic value independent of the rest of variables evaluated (p < 0.0001). CONCLUSIONS: The genetic alterations giving rise to microsatellite instability lead to a better prognosis in patients with colorectal cancer