RESUMEN
Metastatic gastric cancer (mGC) often has a poor prognosis and may benefit from a few targeted therapies. Ramucirumab-based anti-angiogenic therapy targeting the VEGFR2 represents a milestone in the second-line treatment of mGC. Several studies on different cancers are focusing on the major VEGFR2 ligand status, meaning VEGFA gene copy number and protein overexpression, as a prognostic marker and predictor of response to anti-angiogenic therapy. Following this insight, our study aims to examine the role of VEGFA status as a predictive biomarker for the outcome of second-line therapy with Ramucirumab and paclitaxel in mGC patients. To this purpose, the copy number of the VEGFA gene, by fluorescence in situ hybridization experiments, and its expression in tumor tissue as well as the density of micro-vessels, by immunohistochemistry experiments, were assessed in samples derived from mGC patients. This analysis found that amplification of VEGFA concomitantly with VEGFA overexpression and overexpression of VEGFA with micro-vessels density are more represented in patients showing disease control during treatment with Ramucirumab. In addition, in the analyzed series, it was found that amplification was not always associated with overexpression of VEGFA, but overexpression of VEGFA correlates with high micro-vessel density. In conclusion, overexpression of VEGFA could emerge as a potential biomarker to predict the response to anti-angiogenic therapy.
RESUMEN
Lynch syndrome (LS) is an inherited cancer susceptibility syndrome caused by germline mutations in a DNA mismatch repair (MMR) gene or in the EPCAM gene. LS is associated with an increased lifetime risk of colorectal cancer (CRC) and other malignancies. The screening algorithm for LS patient selection is based on the identification of CRC specimens that have MMR loss/high microsatellite instability (MSI-H) and are wild-type for BRAFV600. Here, we sought to clinically and molecularly characterize patients with these features. From 2017 to 2023, 841 CRC patients were evaluated for MSI and BRAFV600E mutation status, 100 of which showed MSI-H. Of these, 70 were wild-type for BRAFV600. Among these 70 patients, 30 were genetically tested for germline variants in hereditary cancer predisposition syndrome genes. This analysis showed that 19 of these 30 patients (63.3%) harbored a germline pathogenic or likely pathogenic variant in MMR genes, 2 (6.7%) harbored a variant of unknown significance (VUS) in MMR genes, 3 (10%) harbored a VUS in other cancer-related genes, and 6 (20%) were negative to genetic testing. These findings highlight the importance of personalized medicine for tailored genetic counseling, management, and surveillance of families with LS and other hereditary cancer syndromes.
RESUMEN
Inflammasomes are multiprotein complexes expressed by immune cells in response to distinct stimuli that trigger inflammatory responses and the release of pro-inflammatory cytokines. Evidence suggests a different role of inflammasome NLRP3 in IBD. NLRP3 inflammasome activation can be controlled by post-translational modifications such as ubiquitination through BRCC3. The aim of this study was to investigate the effect of miR-369-3p on the expression and activation of NLRP3 inflammasomes via BRCC3 regulation. After bioinformatics prediction of Brcc3 as a gene target of miR-369-3p, in vitro, we validated its modulation in bone marrow-derived macrophages (BMDM). The increase in miR-369-3p significantly reduced BRCC3 gene and protein expression. This modulation, in turn, reduced the expression of NLRP3 and blocked the recruitment of ASC adaptor protein by NLRP3. As a result, miR-369-3p reduced the activity of Caspase-1 by the inflammasome, decreasing the cleavage of pro-IL-1ß and pro-IL-18. These results support a novel mechanism that seems to act on post-translational modification of NLRP3 inflammasome activation by BRCC3. This may be an interesting new target in the personalized treatment of inflammatory disorders, including IBD.
Asunto(s)
Enfermedades Inflamatorias del Intestino , MicroARNs , Humanos , Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteínas Adaptadoras Transductoras de Señales , MicroARNs/genética , Enzimas DesubicuitinizantesRESUMEN
Anaplastic classic Kaposi sarcoma (CKS) is an extremely rare pathologic variant of CKS characterized by high aggressiveness and poor prognosis. We report the clinical course of this malignant histologic form in an otherwise healthy 67-year-old male from Apulia in Southern Italy. The anaplastic progression arose during a long history of CKS and developed after multiple local and systemic treatments. The extremely aggressive and chemorefractory nature of the disease dictated amputation of a lower limb and, later, surgery for metastatic pulmonary involvement. At subsequent relapse, therapy with the anti-PD-1 inhibitor pembrolizumab was started. The immunotherapy was selected based on the PD-L1 expression in the tumor and tumor microenvironment. Remarkably, PD-1 blockade induced a complete and durable response in the patient, with a disease-free survival that has exceeded 18 months, and follow-up is still ongoing.
Asunto(s)
Sarcoma de Kaposi , Neoplasias Cutáneas , Masculino , Humanos , Anciano , Sarcoma de Kaposi/terapia , Antígeno B7-H1/uso terapéutico , Neoplasias Cutáneas/patología , Supervivencia sin Enfermedad , Inmunoterapia , Microambiente TumoralRESUMEN
The immunoproteasome is a multi-catalytic protein complex expressed in hematopoietic cells. Increased expression of immuno-subunits followed by increased proteasome activities is associated with the pathogenesis of IBD. Therefore, the identification of molecules that could inhibit the activities of this complex has been widely studied. microRNAs are small molecules of non-coding RNA that regulate the expression of target genes. Our purpose was to demonstrate that miR-369-3p is able to reduce the expression of the PSMB9 subunit and consequently modulate the catalytic activities of immunoproteasome. After bioinformatics prediction of the gene target of miR-369-3p, we validated its modulation on PSMB9 expression in the RAW264.7 cell line in vitro. We also found that miR-369-3p indirectly reduced the expression of other immunoproteasome subunits and that this regulation reduced the catalytic functions of the immunoproteasome. Increased levels of PSMB9 were observed in colon samples of acute IBD patients compared to the remission IBD group and control group. Our data suggest that miR-369-3p may be a future alternative therapeutic approach to several compounds currently used for the treatment of inflammatory disorders including IBD.
Asunto(s)
Enfermedades Inflamatorias del Intestino , MicroARNs , Animales , Humanos , Catálisis , Colon , Enfermedades Inflamatorias del Intestino/genética , Intestinos , MicroARNs/genética , RatonesRESUMEN
SLC15A4/PHT1 is an endolysosome-resident carrier of oligopeptides and histidine recently come into view as a key path marker of immune/autoimmune/inflammatory pathways in immune cells. Yet, its emerging role in inflammatory processes directly targeting the gastrointestinal epithelial layer, as in the multifactorial pathophysiology of inflammatory bowel disease (IBD), is poorly investigated. Here, the first identification of SLC15A4/PHT1 gene products in human colonic epithelium of ulcerative colitis (UC) patients is reported, showing protein primarily localized in intracellular vesicle-like compartments. Qualitative and quantitative immunohistochemical analyses of colon biopsies revealed overexpression of SLC15A4/PHT1 protein product in the epithelial layer of UC patients. Results were successfully mirrored in vitro, in spontaneously differentiated enterocyte-like monolayers of Caco-2 cells specifically exposed to DSS (dextran sodium sulphate) to mimic IBD inflammatory onsets. SLC15A4/PHT1 expression and cellular localization were characterized confirming its (dys)regulation traits in inflamed vs. healthy epithelia, strongly hinting the hypothesis of SLC15A4/PHT1 increased function associated with epithelial inflammation in IBD patients.
Asunto(s)
Colitis Ulcerosa , Enfermedades Inflamatorias del Intestino , Proteínas de Transporte de Membrana , Humanos , Células CACO-2 , Colitis/patología , Colitis Ulcerosa/genética , Colitis Ulcerosa/metabolismo , Colitis Ulcerosa/patología , Colon/metabolismo , Colon/patología , Sulfato de Dextran , Enfermedades Inflamatorias del Intestino/metabolismo , Mucosa Intestinal/metabolismo , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Ratones Endogámicos C57BL , Proteínas del Tejido Nervioso/metabolismo , Regulación hacia ArribaRESUMEN
c-MYC is one of the most important factors involved in colorectal cancer (CRC) initiation and progression; indeed, it is found to be upregulated in up to 80% of sporadic cases. During colorectal carcinogenesis, c-MYC is maintained upregulated through ß-catenin-mediated transcriptional activation and ERK-mediated post-translational stabilization. Our data demonstrate that p38α, a kinase involved in CRC metabolism and survival, contributes to c-Myc protein stability. Moreover, we show that p38α, like ERK, stabilizes c-MYC protein levels by preventing its ubiquitination. Of note, we found that p38α phosphorylates c-MYC and interacts with it both in vitro and in cellulo. Extensive molecular analyses in the cellular and in vivo models revealed that the p38α kinase inhibitors, SB202190 and ralimetinib, affect c-MYC protein levels. Ralimetinib also exhibited a synthetic lethality effect when used in combination with the MEK1 inhibitor trametinib. Overall, our findings identify p38α as a promising therapeutic target, acting directly on c-MYC, with potential implications for countering c-MYC-mediated CRC proliferation, metastatic dissemination, and chemoresistance.
RESUMEN
Genetic variants located in non-coding regions can affect processes that regulate protein expression, functionally contributing to human disease. Germline heterozygous mutations in the non-coding region of the PTEN gene have been previously identified in patients with PTEN hamartoma tumor syndrome (PHTS) diagnosed with breast, thyroid, and/or endometrial cancer. In this study, we report a PTEN promoter variant (rs34149102 A allele) that was identified by direct sequencing in an Italian family with a history of gastroesophageal junction (GEJ) adenocarcinoma and breast cancer. In order to investigate the putative functional role of the rs34149102 A allele variant, we evaluated the status of PTEN alterations at the somatic level. We found that PTEN protein expression was absent in the GEJ adenocarcinoma tissue of the index case. Moreover, we detected the occurrence of copy number loss involving the PTEN rs34149102 major C allele in tumor tissue, revealing that the second allele was somatically inactivated. This variant is located within an active regulatory region of the PTEN core promoter, and in silico analysis suggests that it may affect the binding of the nuclear transcription factor MAZ and hence PTEN expression. Overall, these results reveal the functional role of the PTEN promoter rs34149102 A allele variant in the modulation of PTEN protein expression and highlight its contribution to hereditary cancer risk.
Asunto(s)
Adenocarcinoma , Neoplasias de la Mama , Síndrome de Hamartoma Múltiple , Neoplasias de la Mama/genética , Neoplasias Esofágicas , Femenino , Células Germinativas/metabolismo , Síndrome de Hamartoma Múltiple/genética , Humanos , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/metabolismoRESUMEN
To identify useful markers for prognostic and therapeutic purposes, The Cancer Genome Atlas (TCGA) provided a molecular classification of gastric cancers (GCs). Previous studies have used immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) to define immunophenotypic surrogate markers of the molecular alterations. Some critical issues concerning the correct definition of immunophenotypic groups have emerged in these studies that employed tissue microarrays (TMAs). We performed an immunophenotypic classification by evaluating MLH1, p53, HER2, E-cadherin, and Epstein-Barr virus (EBV) on the whole section of the surgical GC samples compared to most of the studies conducted on TMAs. We also investigated the immunohistochemical expression of PD-L1, a known therapeutic target. We identified the following immunophenotypic groups: EBV (2.9%); mismatch repair deficient (MMR-D) (7.2%); overexpressed p53 and/or HER2+ (61.4%); aberrant E-cadherin (11.4%); and normal pattern (17.1%). The use of surgical samples emphasized that some immunohistochemical markers were not useful for properly classifying the GC specimens. We can state that EBV (significantly correlated to PD-L1 expression) and MMR-D GCs are well-defined groups, mutually exclusive, and easily assessable with IHC and CISH, and could be candidates for immunotherapy with PD-1/PD-L1 inhibitors. As regards p53, our findings suggest that IHC assessment may be responsible for a misclassification of GC groups. Immunohistochemical evaluation of E-cadherin needs to be standardized, particularly in terms of the heterogeneous cytoplasmic/membranous staining pattern. Whether to consider the normal-pattern group as a separate category remains to be clarified. Because GC specimens with known therapeutic targets account for only 40%, we suggest reviewing the immunophenotypic classification to find new therapeutic targets, such as PD-L1, MLH1, and HER2.
Asunto(s)
Inmunohistoquímica/métodos , Fenotipo , Neoplasias Gástricas/clasificación , Neoplasias Gástricas/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Antígeno B7-H1/metabolismo , Biomarcadores de Tumor/metabolismo , Infecciones por Virus de Epstein-Barr/metabolismo , Infecciones por Virus de Epstein-Barr/virología , Femenino , Herpesvirus Humano 4/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Homólogo 1 de la Proteína MutL/metabolismo , Pronóstico , Receptor ErbB-2/metabolismo , Neoplasias Gástricas/patologíaRESUMEN
The treatment of patients with human epidermal growth factor receptor 2 (HER2)-negative gastric cancer is a major challenge. Immunotherapy using immune checkpoint inhibitors is a rapidly growing field. In a number of malignancy types it has been demonstrated that patients with mismatch repair deficiency efficiently respond to programmed death-ligand 1 (PD-L1) blockade therapy. Recent studies have evaluated tumor microenvironment immune types to predict which patients may clinically benefit from immunotherapy. The present study aimed to evaluate the immunohistochemical expression of PD-L1 in 70 gastric cancer tissue samples. Potential associations between PD-L1 expression and mismatch repair deficiency, HER2 and Epstein Barr virus (EBV) status were then investigated in the context of the tumor microenvironment. A positive association was identified for PD-L1 expression with mismatch repair deficiency and EBV status; however, no association was revealed with HER2 status. Immunohistochemistry was then used to classify the microenvironment immune types. This demonstrated that the majority of the gastric cancer samples (73%) belonged to the tumor microenvironment immune type II [PD-L1-/cluster of differentiation 8 (CD8)+ low], which involves an immune ignorant state and has a low sensitivity to immunotherapy. However, 7% of the gastric cancer cases were identified to belong to the tumor microenvironment immune type I (PD-L1+/CD8+ high), which exhibits adaptive immune escape responses and a high chance of reversion with immune checkpoint blockade therapy. In conclusion, the present study emphasized the importance of evaluating tumor microenvironment immune types, mismatch repair deficiency status and EBV status, rather than PD-L1 expression alone, when evaluating the eligibility of a patient for immunotherapy with anti-programmed cell death protein-1/PD-L1 antibodies.
RESUMEN
RATIONALE: Inflammatory bowel disease (IBD) patients manifest symptoms of disturbed gut function, such as neural sensory-motor changes. Programmed cell death-ligand 1 (PD-L1), normally present in neural tissue, exists in close apposition to the mucosal immune system and intestinal epithelium, and a bi-directional communication is known to occur at these interfaces. Somatostatin has been shown to suppress the inflammatory reaction, and has been used in several clinical trials to treat inflammatory disorders, such rheumatoid arthritis. Recently, somatostatin receptor type 2A, that regulates neurotransmission, proliferation, and apoptosis, has been recognized in IBD. Although prominent abnormalities in the morphology of the enteric nervous system have been observed in idiopathic IBD, they are more marked in Crohn disease. PATIENT CONCERNS: A 55-year-old woman with recurrent Crohn disease, just surgically treated for ileal resection, have a stenotic complication. INTERVENTIONS: At surgery 5âcm of preterminal ileum with stenosis and anastomotic ileocolic block was removed. DIAGNOSES: The histopathology showed a recurrent Crohn in fistulo-stenotic phase; the stenosis was mainly sustained by mass-forming, ganglioneuromatous hyperplasia. Normally very rare, fine nerve twigs extend up into mucosa but we found a new-formed fibrillary network, extending into the inflammation area at the subepithelial luminal site of the mucosa, that was positive to PD-L1 and somatostatin receptor type 2A (SSTR2A) immunostaining but not visualized in routinary stained slides. OUTCOMES: After surgery the patient was semestrally followed with clinical endoscopic evaluation that results uneventfully. LESSONS: Our case shows that before surgery neuromatous abnormalities can be predicted by immunostained new-formed twigs in the mucosa.
Asunto(s)
Antígeno B7-H1/metabolismo , Enfermedad de Crohn/patología , Mucosa Intestinal/patología , Intestino Delgado/patología , Receptores de Somatostatina/metabolismo , Enfermedad de Crohn/metabolismo , Enfermedad de Crohn/cirugía , Femenino , Humanos , Mucosa Intestinal/inervación , Mucosa Intestinal/metabolismo , Mucosa Intestinal/cirugía , Intestino Delgado/inervación , Intestino Delgado/metabolismo , Intestino Delgado/cirugía , Persona de Mediana Edad , RecurrenciaRESUMEN
OBJECTIVES: We investigated the PD-L1 expression in colorectal cancer (CRC) and in its microenvironment. RESULTS: PD-L1 was expressed in neoplastic cells (NCs) and tumor-infiltrating immune cells (IICs). All samples PD-L1+ on NCs were also on IICs. Three types of cancers could be grouped: group A(NCs-/ IICs-); group B (NCs-/ IICs+); group C (NCs+/IICs+). To group A belong tumors characterized by poorly immunogenic competence, poor immune response but massive granulocyte infiltrate, justifying the absence of PD-L1 as an immunoinhibitor receptor. To Group B probably belong more immunogenic CRCs, justifying the strong IICs-mediated immune response, and up-regulation of PD-L1 expression only on IICs. To group C belong CRCs probably characterized by a large amount of tumor neoantigens resulting in a marked infiltration of lymphocytes and PD-L1 upregulation also in NCs. MATERIALS AND METHODS: Sixty-three colorectal cancer specimens from a cohort of 61 patients were retrospectively reviewed. Thirty-seven MSS and 26 MSI-H CRCs enrolled in this study. Immunohistochemical staining to PD-L1 was performed by using MAb E1L3N. CONCLUSIONS: Our study calls attention to the importance to assess PD-L1 expression in tumor microenvironment also evaluating type and density of infiltrating immune cells to better stratify CRCs with different immunological patterns.
RESUMEN
Neuroendocrine neoplasms (NENs) are rare, heterogeneous and ubiquitous tumors commonly localized in the gastrointestinal tract, lung, and pancreas. The clinical behavior of NEN is highly unpredictable; in fact, low-grade cases can unexpectedly be associated with metastases. Currently, the 2010 WHO NEN classification employs histological differentiation and the proliferation index for grading tumors but fails to provide reliable prognostic and therapeutic indications. Therefore, there is an urgent need for a better characterization of G2/G3 NENs. Similar to several other tumors, NENs possess immune-escape mechanisms, but very little has yet been done to characterize this crucial aspect. There are no available data describing PD-L1 expression in these tumors. Here we provide, for the first time, evidence of PD-L1 tissue expression in gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs). PD-L1 expression was significantly associated with a high-grade WHO classification (G3) (P<0.001) but not with gender, primary site, or lymph node status. The PD-L1 positivity rate and signal intensity are directly correlated (P<0.001) with a grade increase from G1 to G3. In particular in G3 cases, we observed a dichotomy between the morphology (WD- and PD-NENs) and Ki67. Moreover, our study demonstrated a significant association with the grade and PD-L1 expression levels in immune-infiltrating cells (P<0.001). In particular, G3 tumors are characterized by strong PD-L1 expression in both the tumor and infiltrating immune cells (P<0.001), reflecting an unfavorable environment for T-cell-mediated tumor aggression. These findings suggest that NENs might acquire resistance to immune surveillance by upregulating PD-L1 and inhibiting peritumoral and intratumoral infiltrating lymphocytes. Here we demonstrate that PD-L1 is currently the best-known biomarker for G3 NENs, becoming the new gold standard for G3 NEN discrimination. Furthermore, pharmacological approaches using anti-PD-1 antibodies may become the logical choice for the treatment of G3 cases with a poor prognosis.
Asunto(s)
Antígeno B7-H1/biosíntesis , Biomarcadores de Tumor/biosíntesis , Neoplasias Intestinales/metabolismo , Tumores Neuroendocrinos/metabolismo , Neoplasias Pancreáticas/metabolismo , Neoplasias Gástricas/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Neoplasias Intestinales/patología , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Tumores Neuroendocrinos/patología , Neoplasias Pancreáticas/patología , Neoplasias Gástricas/patologíaRESUMEN
BACKGROUND: Myocardial infarction and colorectal cancer are associated at the population level and in autoptic studies. Glycated apolipoprotein B (apoB) is a risk factor for the development of myocardial infarction. The association of glycated apoB with dysplastic and neoplastic colorectal tissue was investigated. MATERIALS AND METHODS: Forty-eight consecutive surgical specimens, 26 colorectal adenomas and 22 colorectal carcinomas, retrieved from the archives of the Pathologic Anatomy Department of our institution, were examined. The tissue content of glycated apoB was determined using a monoclonal antibody. RESULTS: Glycated apoB was detected in 27% of the adenomas and 45% of the cancer tissue, but only in 18% of the normal tissue near the cancer site. CONCLUSION: Glycated apoB is associated with dysplastic and even more so with neoplastic cancer tissue.
Asunto(s)
Adenoma/metabolismo , Pólipos Adenomatosos/metabolismo , Neoplasias Colorrectales/metabolismo , Lipoproteínas LDL/biosíntesis , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Adenoma/patología , Pólipos Adenomatosos/patología , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/patología , Femenino , Productos Finales de Glicación Avanzada , Humanos , Inmunohistoquímica , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND/AIM: The crucial role of KRAS status in new colorectal cancer target therapy raises the issue regarding which testing method to use. This study analysed 112 formalin fixed, paraffin-embedded (FFPE) metastatic tissue samples using three different commercially available kits. PATIENTS AND METHODS: A group of 40 KRAS wild-type (wt), 40 codon 12-mutated and 32 codon-13 mutated samples, previously evaluated by real-time PCR (TheraScreen kit), used as reference method, were analysed by Ampli-set-K-RAS and K-RAS StripAssay kit (herein called kit A and B, respectively) based on two different technologies. RESULTS: The sensitivity of both kits was 92.5% for wt samples, 100% and 95.0% for kit A and B, respectively for samples mutated in codon 12. The specificity was 100% for both kits for all groups of samples. After a minor modification of the kit A method, its specificity reached 100%. CONCLUSION: of low cost and easy to use, kit A may be suitable for use in a routine diagnostic setting.
Asunto(s)
Biomarcadores de Tumor/genética , Neoplasias Colorrectales/genética , Genes ras , Tiras Reactivas , Codón , Neoplasias Colorrectales/patología , ADN de Neoplasias/análisis , ADN de Neoplasias/genética , Genotipo , Humanos , Mutación , Metástasis de la Neoplasia , Adhesión en Parafina , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y EspecificidadRESUMEN
The aims of this study were: (i) to examine frozen-thawed ovarian tissues for features of follicular health and atresia by histology; (ii) to assess the expression of estrogen receptors alpha (ERalpha) and beta (ERbeta) by real-time PCR; (iii) to evaluate the Bax/Bcl-2 ratio, as an apoptotic index, in the ovarian tissues before and after cryopreservation. Ovarian cortical biopsies were obtained from 11 patients. The fragments were subdivided into two groups, fresh (control tissues) and cryopreserved tissues obtained by direct plunging into liquid nitrogen. Both tissue groups were subjected to a histological evaluation of the healthy and atretic follicles, immunohistochemical localization of the ER, and a real-time PCR (qPCR) to evaluate the expression of ER, Bax, Bcl-2 as well as beta-actin, as control gene. Damage was observed in 31% of primordial, 45% of primary, and 75% of secondary follicles in the cryopreserved tissue group. The qPCR analysis showed that the level of ERbeta was greater in fresh than cryopreserved tissues, whereas the ERalpha expression and Bax/Bcl-2 ratio were similar in both tissue groups. A significant inverse association was observed between ERalpha mRNA levels in the fresh tissue group and subjects' ages. The results show that cryopreservation and thawing of human ovarian tissue does not affect the morphology of primordial or primary follicles and that cryopreservation does not affect apoptosis. However, cryopreservation seems to have an inhibitory effect on the level of ERbeta. Additional studies are needed to evaluate the differential effects of freezing follicles at different stages of follicular development and ovarian steroidogenesis.
Asunto(s)
Apoptosis/fisiología , Criopreservación , Receptor alfa de Estrógeno/metabolismo , Receptor beta de Estrógeno/metabolismo , Folículo Ovárico/patología , Ovario/metabolismo , Ovario/patología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Adolescente , Adulto , Femenino , Humanos , Folículo Ovárico/metabolismo , ARN Mensajero/metabolismoRESUMEN
The EGF receptor (EGFR) has emerged as a rational target for anticancer therapy for the treatment of colorectal cancer (CRC). Positive immunohistochemistry (IHC) staining for EGFR is used as a criterion for patient selection; however, doubt has been cast on the utility of this method. Not only is the response to cetuximab, an anti-EGFR mAb, low in patients expressing EGFRs, but a similar response to cetuximab has also been described in patients who do not express EGFRs. This review aims to evaluate the possible cause of the lack of correlation between the efficacy of cetuximab and EGFR IHC staining in CRC, as well as any modifications in the IHC method necessary to optimize patient selection for cetuximab therapy. In our opinion, the heterogeneous expression of the receptor in the neoplastic population and the inability of the mAbs used to predict the response to cetuximab could be the major cause of the failure of IHC staining as a reliable tool for patient selection. The use of specific mAbs directed against the phosphorylated and mutant form (EGFRvIII) of the EGFR could reinstate IHC as a valid predictor of response to therapy.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Neoplasias Colorrectales/tratamiento farmacológico , Receptores ErbB/efectos de los fármacos , Anticuerpos Monoclonales Humanizados , Cetuximab , Humanos , Inmunohistoquímica , Hibridación in SituRESUMEN
Whereas long-term cholestasis results in intestinal alterations and increased permeability to hepatotoxins, the effect of short-term cholestasis is less known and was investigated in bile duct ligated (BDL) rats. In the intestinal mucosa, at Day 7 BDL, total glutathione and protein sulfhydryl contents had decreased, oxidized glutathione levels increased (P<0.05 vs baseline), and a reduced epithelium thickness with dissolving crypt phenomena was observed in 40% of rats. At Day 10, total protein content, glutathione-related enzyme activities, and the transmural electrophysiological activity had decreased (-50%); by contrast, oxidized proteins doubled (P<0.05), and histological changes were extended to 70% of rats. In vitro exposure to taurodeoxycholate at micellar concentrations determined dysepithelization in normal gut but dissolving crypt phenomena and necrosis in cholestatic bowels. In the liver, ongoing cholestasis was associated with early oxidative changes especially in mitochondria, where protein sulfhydryls were decreased and negatively correlated with glutathione-protein mixed disulfides (r=-0.807, P<0.001). Daily oral administration of tauroursodeoxycholate, a hydrophilic bile salt, and glutathione to BDL rats improved intestinal histology, function, and redox state. In conclusion, short-term cholestasis results in distinctive functional, oxidative, and morphological changes of intestinal mucosa, determined increased vulnerability to toxic injury, and parallel hepatic oxidative damage.
Asunto(s)
Antioxidantes/uso terapéutico , Ácidos y Sales Biliares/uso terapéutico , Colestasis/complicaciones , Intestinos/lesiones , Hígado/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Animales , Conductos Biliares/fisiología , Modelos Animales de Enfermedad , Disulfuro de Glutatión/metabolismo , Intestinos/efectos de los fármacos , Hígado/lesiones , Masculino , Ratas , Ratas WistarRESUMEN
Mismatch repair (MMR) proteins are capable of recognizing and processing not only single base-pair mismatches and insertion-deletion loops that occur during DNA replication, but also adducts in DNA resulting from treatment with cancer chemotherapy agents. MMR deficiency leads to microsatellite instability (MSI) and results in resistance to antimetabolites, alkylating and platinating agents, DNA minor groove binders, and inhibitors of topoisomerases. Therefore, anticancer agents that can be recommended for use in MMR deficient colorectal cancers are those that exert their cytotoxicity regardless of the MMR status. These include some alkylating drugs, brostacillin, gemcytabine, photodynamic therapy, taxanes. An approach that is currently receiving much attention is the use of agents such as 5-azacytidine, an inhibitor of the DNA methyltransferases, in combination with inhibitors of histone de-acetylation, to restore the MMR function. A strong anti-proliferative efficacy with a relatively low direct cytotoxicity, obtainable with oloumicine and roscovitine (selective cyclin-dependent kinases inhibitors) can represent a new expedient for the therapeutic treatment of MMR deficient colorectal cancers. The question of how MMR defects modulate the response to chemotherapeutics deserves further investigation, to enable a more aware choice of cancer treatment.
Asunto(s)
Antimetabolitos Antineoplásicos/uso terapéutico , Azacitidina/uso terapéutico , Disparidad de Par Base/genética , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Humanos , Estadificación de NeoplasiasRESUMEN
Immunohistochemical (IHC) assessment of the mismatch repair proteins has been proposed as an alternative strategy to molecular biology for evaluating the unstable phenotype of tumors. With the aim of introducing IHC analysis as a routine diagnostic test, the authors compared the expression of MLH1 and MSH2 proteins with a PCR-based microsatellite assay. The concordance rate between the two methods was 90% after IHC evaluation of two different areas of each tumor. These results show that IHC may be as efficient as PCR in detecting unstable phenotype by using only one surgical or biopsy sample.