Study of long non-coding RNA Tug 1 expression in Egyptian colorectal adenocarcinoma patients

Purpose: Colorectal cancer (CRC) is one of the most fatal tumors worldwide. In Egypt, most CRC cases occur in individuals > 40 years old. TUG1 has been proved to be disrupted in different malignancies and may have a critical role in tumor progression, invasion, and metastasis. However, its role in CRC has not been adequately studied. Materials / Methods: Quantitative real-time polymerase chain reaction (PCR) was used to evaluate the expression levels of long non-coding RNA (LncRNA) taurine upregulated gene 1 (TUG1), in nonmetastatic and metastatic CRC tissues and adjacent noncancerous tissues as control. Results: LncRNA TUG1 expression was significantly upregulated in both nonmetastatic and metastatic CRC tissues, in comparison with the adjacent noncancerous tissue. It was found that TUG1 could have a possible prognostic role in CRC, by comparing the sensitivity and specificity of TUG1 with those of CEA and CA19-9. Conclusion: The results of the current study suggest that the LncRNA TUG1 participates in the malignant behaviors of CRC cells. (AU)

Data mining identifies novel RNA-binding proteins involved in colon and rectal carcinomas.

Colorectal adenocarcinoma (COREAD) is the second most deadly cancer and third most frequently encountered malignancy worldwide. Despite efforts in molecular subtyping and subsequent personalized COREAD treatments, multidisciplinary evidence suggests separating COREAD into colon cancer (COAD) and rectal cancer (READ). This new perspective could improve diagnosis and treatment of both carcinomas. RNA-binding proteins (RBPs), as critical regulators of every hallmark of cancer, could fulfill the need to identify sensitive biomarkers for COAD and READ separately. To detect new RBPs involved in COAD and READ progression, here we used a multidata integration strategy to prioritize tumorigenic RBPs. We analyzed and integrated 1) RBPs genomic and transcriptomic alterations from 488 COAD and 155 READ patients, 2) ∼ 10,000 raw associations between RBPs and cancer genes, 3) ∼ 15,000 immunostainings, and 4) loss-of-function screens performed in 102 COREAD cell lines. Thus, we unraveled new putative roles of NOP56, RBM12, NAT10, FKBP1A, EMG1, and CSE1L in COAD and READ progression. Interestingly, FKBP1A and EMG1 have never been related with any of these carcinomas but presented tumorigenic features in other cancer types. Subsequent survival analyses highlighted the clinical relevance of FKBP1A, NOP56, and NAT10 mRNA expression to predict poor prognosis in COREAD and COAD patients. Further research should be performed to validate their clinical potential and to elucidate their molecular mechanisms underlying these malignancies.

Combination of vitamin D and probiotics inhibits chemically induced colorectal carcinogenesis in Wistar rats.

The modulation of inflammatory elements, cell differentiation and proliferation by vitamin D and the role of probiotics in the intestinal microbiota and immunogenic response have sparked interest in the application of both in chemotherapeutics and chemoprevention of colorectal tumors. AIMS: The present study aimed to investigate the effects of isolated and/or combined treatment of vitamin D3 and probiotics on colorectal carcinogenesis. MAIN METHODS: Pre-neoplastic lesions were induced with 1,2-dimethylhydrazine in the colon of Wistar rats, which were treated with probiotics and/or vitamin D in three different approaches (simultaneous, pre-, and post-treatment). We investigated the frequency of aberrant crypt foci (ACF) and aberrant crypt (AC) in the distal colon, fecal microbiome composition, gene and protein expression through immunohistochemical and RT-PCR assays, and general toxicity through water consumption and weight gain monitoring. KEY FINDINGS: Results confirm the systemic safety of treatments, and show a protective effect of vitamin D and probiotics in all approaches studied, as well as in combined treatments, with predominance of different bacterial phyla compared to controls. Treated groups show different levels of Nrf2, GST, COX2, iNOS, ß-catenin and PCNA expression. SIGNIFICANCE: These experimental conditions explore the combination of vitamin D and probiotics supplementation at low doses over pathways involved in distinct stages of colorectal carcinogenesis, with results supporting its application in prevention and long-term strategies.

Tissue factor expression in colorectal adenocarcinoma: association with angiogenesis and clinical and pathological aspects

Introduction: Tissue factor (TF) expression has been described in various neoplasms and was correlated with angiogenesis and metastases. Objectives: To describe TF expression in colorectal cancers, correlating it with microvessel density and clinical and pathological variables. Methods: Immunohistochemistry was used to determine TF expression and microvessel density. The Student t-test was used to compare high and low TF expression with microvessel density andwith age. The chi-squared test was used for other comparisons, and Kaplan-Meier curves were used for survival analyses. Results: Forty-three patients were operated with curative intent. Their mean age was 58.1±12.6 years old, and 62.8% were male. The rectum was the most common location (60,4%), and most tumors reached the serosa and peri-intestinal fat (72.1%). Lymph nodes were positive in 46.5%, and 72.1% of the tumors were moderately differentiated adenocarcinomas. Death occurred in 27.6±12.8months in 51.1% of the patients who had recurrence. Tissue factor expression was intense in 88.4%. There was a positive correlation between TF expression and microvessel density (p=0.02), and between TF and older age (p< 0.01). There was no correlation between TF expression and other variables (gender, histological type, penetration into the intestinal wall, and lymphatic and systemic metastases). Tissue factor expression did not correlate with survival. Conclusion: Tissue factor expression correlated with increased microvessel density and older age. Further studies are necessary to ascertain the clinical relevance of TF in colorectal cancer. (AU)

Profiles of endoglin and vascular endothelial growth factor based on staging and histological grading of colorectal cancer and their relationship with bevacizumab therapy / Perfis da endoglina e do fator de crescimento endotelial vascular com base no estadiamento e na graduação histológica do câncer colorretal e sua relação coma terapia com bevacizumabe

Objective: The present study evaluated the profile of endoglin (CD105) and vascular endothelial growth factor (VEGF) based on staging and histopathological grading of colorectal cancer as well as their relationship with bevacizumab therapy. Methods: A total of 88 cases of colorectal adenocarcinoma were included in the present study. The levels of VEGF and CD105 protein were evaluated with enzymelinked immunosorbent assay (ELISA). Results: There was a significant difference in the level of CD105 (p=0.002) between metastases and non-metastases subjects, showing that CD105 was higher in metastases subjects (4.59 ng/ml). Therewas no significant difference in the level of VEGF based on the presence of metastasis (p=0.625). There was a significant difference in the levels of CD105 (p=0.038) and VEGF (p=0.010) between the subjects who received chemotherapy and those who did not. The CD105 level was higher in the subjects who received chemotherapy (4.43 ng/ml); conversely, the level of VEGF was lower in subjects who received chemotherapy (543.65 pg/ml). There was a statistically significant difference in the levels of CD105 (p=0.003) and VEGF (p=0.002) between subjects who received bevacizumab therapy and subjects who did not. The levels of CD105 were higher in subjects who received bevacizumab therapy (5.11 ng/ml); in contrast, the level of VEGF was higher in subjects who did not receive bevacizumab therapy (645.92 pg/ml). There was a significant positive correlation between CD105 and VEGF in subjects who did not receive bevacizumab (p<0.01). Conclusion: The results of this study support a hypothesis of "escape mechanism" in the failure of anti-angiogenesis therapy (anti-VEGF). (AU)

Objetivo: Este estudo avaliou o perfil da endoglina (CD105) e do fator de crescimento endotelial vascular (FCEV) com base no estadiamento e graduação histopatológica do câncer colorretal, assim como sua relação com a terapia com bevacizumabe. Métodos: No total, 88 casos de adenocarcinoma colorretal foram incluídos no presente estudo. Os níveis das proteínas FCEV e CD105 foram avaliados com ensaio imunoenzimático (ELISA, na sigla em inglês). Resultados Houve uma diferença significativa no nível de CD105 (p=0,002) entre indivíduos commetástases e semmetástases, que indicou que o nível de CD105 émais alto em indivíduos com metástases (4,59 ng/ml). Não houve diferença significativa no nível de FCEV com base na presença de metástases (p=0,625). Houve diferença significativa nos níveis de CD105 (p=0,038) e de FCEV (p=0,010) entre os indivíduos que receberam quimioterapia e os que não receberam. Encontrou-se um nível de CD105 mais alto nos indivíduos que submetidos a quimioterapia (4,43 ng/ml); Em contrapartida, encontrou-se um nível de FCEV mais baixo em indivíduos que submetidos a quimioterapia (543,65 pg/ml). Houve uma diferença estatisticamente significativa nos níveis de CD105 (p=0,003) e de FCEV (p=0,002) entre os indivíduos submetidos e não submetidos à terapia com bevacizumabe. Os níveis de CD105 foram mais elevados em indivíduos submetidos à terapia combevacizumab (5,11 ng/ml); em contraste, observou-se um nível de FCEV mais alto em indivíduos que não foram submetidos à terapia com bevacizumabe (645,92 pg/ml). Houve uma correlação positiva significativa entre CD105 e FCEV em indivíduos que não receberam bevacizumabe (p<0.01). Conclusão: Os resultados deste estudo corroboram a hipótese de "mecanismo de escape" na falha da terapia anti-angiogênica (anti-FCEV). (AU)

Tumor microenvironment-associated gene C3 can predict the prognosis of colorectal adenocarcinoma: a study based on TCGA.

BACKGROUND: Colorectal cancer is one of the most common malignancies. With continuous exploration of the interaction between tumor cells and the immune system, tumor immunotherapy has become a revolution. However, CRC remains one of the less effective tumors for immunotherapy. The tumor microenvironment plays an important role in tumorigenesis and progression. The aim of this study is to explore tumor microenvironment-related genes that can predict the prognosis of colorectal adenocarcinoma, and also to provide new ideas for the mechanism of tumor development as well as immunotherapy. METHODS: After estimating Stromalscore and Immunescore of colorectal adenocarcinoma tumor samples according to RNA-Seq expression data downloaded from TCGA, we screened for TME-related differential genes. We filtered prognosis-related core genes by constructing protein-protein interaction networks and making one-factor cox analysis for prognosis. Finally, the relative content of 22 immune cells in tumor tissues was evaluated, and then immune cells associated with core genes were identified. RESULTS: We screened 773 differential genes related to the TME. Then we identified C3 as a core gene associated with prognosis. Single gene analysis showed that C3 expression was significantly higher in tumor tissues than in normal tissues (p < 0.001). High C3 expression was associated with lower overall survival (p = 0.046). Tumor immune cell analysis showed that mast cells resting, mast cells activated, T cells CD4 memory activated, eosinophils, and macrophages M0 were C3-associated immune cells. CONCLUSIONS: C3 has potential as a biomarker for colorectal adenocarcinoma and could provide new research ideas for the diagnosis and treatment of colorectal adenocarcinoma, especially for immunotherapy.

Resecciones colorrectales segmentarias en pacientes con colitis ulcerosa y displasia o adenocarcinoma / Segmental colectomy in patients with colitis-associated neoplasia or dysplasia

Resumen Introducción: El tratamiento estándar para los pacientes con colitis ulcerosa y displasia o adenocarcinoma de colon ha sido la proctocolectomía total, lo que conlleva una morbilidad y una reducción en la calidad de vida significativa. Materiales y Método: Se hace un análisis retrospectivo de 5 pacientes con colitis ulcerosa a los que se realiza una resección segmentaria por displasia o adenocarcinoma. Resultados: La mediana de edad al diagnóstico de colitis ulcerosa y de la neoplasia fue de 56 y 62 años respectivamente. El tiempo de evolución de la enfermedad fue de 1 a 13 años. La mediana de seguimiento postoperatorio fue de 57 meses apareciendo en uno de los pacientes un nuevo foco de displasia. Conclusiones: En determinados pacientes seleccionados, las resecciones segmentarias podrían ser una opción segura si tienen buen control de la enfermedad, escasa actividad inflamatoria, pocos años de evolución y que puedan realizar un adecuado seguimiento posterior.

Introduction: The historical management for patients with ulcerative colitis and displasia or adenocarcinoma associated was to perform a total proctocolectomy, what cause important morbidity and affect patient's quality of life. Materials and Method: A retrospective review about 5 patients with a segmental colectomy due to dysplasia or adenocarcinoma is done. Results: The median age at diagnosis of ulcerative colitis and cancer was 56 and 62 years respectively. Disease time evolution was between 1 and 13 years. The patients were followed up for a median of 57 months. During the follow-up evaluation, 1 patient was found to have dysplasia. Conclusion: Segmental colectomy could be a safe option in clinically stable patients, few years of diagnosis and with and a suitable follow up.

Biological and toxicological evaluation of Rhus trilobata Nutt. (Anacardiaceae) used traditionally in mexico against cancer.

BACKGROUND: Rhus trilobata Nutt. (Anacardiaceae) (RHTR) is a plant of Mexico that is traditionally used as an alternative treatment for several types of cancer. However, the phytochemical composition and potential toxicity of this plant have not been evaluated to support its therapeutic use. Therefore, this study aimed to evaluate the biological activity of RHTR against colorectal adenocarcinoma cells, determine its possible acute toxicity, and analyze its phytochemical composition. METHODS: The traditional preparation was performed by decoction of stems in distilled water (aqueous extract, AE), and flavonoids were concentrated with C18-cartridges and ethyl acetate (flavonoid fraction, FF). The biological activity was evaluated by MTT viability curves and the TUNEL assay in colorectal adenocarcinoma (CACO-2), ovarian epithelium (CHO-K1) and lung/bronchus epithelium (BEAS-2B) cells. The toxicological effect was determined in female BALB/c mice after 24 h and 14 days of intraperitoneal administration of 200 mg/kg AE and FF, respectively. Later, the animals were sacrificed for histopathological observation of organs and sera obtained by retro-orbital bleeding for biochemical marker analysis. Finally, the phytochemical characterization of AE and FF was conducted by UPLC-MSE. RESULTS: In the MTT assays, AE and FF at 5 and 18 µg/mL decreased the viability of CACO-2 cells compared with cells treated with vehicle or normal cells (p ≤ 0.05, ANOVA), with changes in cell morphology and the induction of apoptosis. Anatomical and histological analysis of organs did not reveal important pathological lesions at the time of assessment. Additionally, biochemical markers remained normal and showed no differences from those of the control group after 24 h and 14 days of treatment (p ≤ 0.05, ANOVA). Finally, UPLC-MSE analysis revealed 173 compounds in AE-RHTR, primarily flavonoids, fatty acids and phenolic acids. The most abundant compounds in AE and FF were quercetin and myricetin derivates (glycosides), methyl gallate, epigallocatechin-3-cinnamate, ß-PGG, fisetin and margaric acid, which might be related to the anticancer properties of RHTR. CONCLUSION: RHTR exhibits biological activity against cancer cells and does not present adverse toxicological effects during its in vivo administration, supporting its traditional use.

Cáncer colorectal en la edad pediátrica: reporte de un caso

El cáncer colorrectal (CCR) constituye una dolencia extremadamente infrecuente en niños. Es más excepcional cuando ocurre en niños con neurofibromatosis tipo 1, que es considerada como factor de riesgo para el desarrollo de tumores malignos. Se reporta el caso de un niño de 8 años con CCR metastático de alto grado. El paciente presentó diarrea crónica muco-sanguinolenta, rectorragia y pérdida de peso. Su examen reveló manchas café con leche diseminadas sobre la espalda, pelvis y miembros inferiores. En la colonoscopia se visualizó una masa polipoide, estenosante, pediculada, ulcerativa, localizada a 35 cm del margen anal. La TAC reveló una metástasis hepática. Se realizó una colectomía amplia y el estudio histológico demostró 23 pólipos, 112 ganglios reaccionales y un tumor con componente mucinoso. Recibió quimioterapia y lobectomía hepática. No hay protocolo específico para el tratamiento de esta afección en niños. El pronóstico depende principalmente del carácter agresivo del tumor, de su estadificación y del lugar en el que se desarrolla.

Colorectal cancer (CRC) is an extremely rare condition in children. It is more exceptional when it oc-curs in children with neurofibromatosis type 1, which is considered a risk factor for the development of malignant tumors. We report the case of an 8-year-old child with high-grade metastatic CRC. The patient had chronic muco-bloody diarrhea, rectal bleeding and weight loss. His examination revealed cafe-au-lait spots scattered on his back, pelvis, and lower limbs. A polipoid, stenosing, pediculated, ul-cerative mass located 35 cm from the anal margin was visualized by colonoscopy. CT revealed hepatic metastasis. An ample colectomy was performed and the histology showed 23 polyps, 112 reactive lymph nodes and one tumor with a mucinous component. The patient received chemotherapy and liver lobectomy. There is no specific protocol for the treatment of this condition in children. The prog-nosis depends mainly on the aggressive nature of the tumor, its staging and the place where it develops.

Expressão das metaloproteinases 2 e 9 em adenocarcinoma colorretal / Metaloproteinases 2 and 9 expression in colorectal adenocarcinoma

Introdução: Os carcinomas da região colorretal representam o terceiro tipo mais comum de neoplasia maligna e a terceira causa de mortalidade relacionada ao câncer no mundo. Objetivos: Este estudo teve como objetivo verificar se existe associação entre estadiamento, invasão vascular, tamanho do tumor, classificações Duke's, Astler-Coller, grau de diferenciação tumoral e expressão das metaloproteinases 2 e 9 em células de adenocarcinoma intestinal. Métodos: Foi realizado um estudo retrospectivo de 40 blocos de parafina contendo amostras obtidas de ressecção cirúrgica de tecido intestinal anteriormente diagnosticado como adenocarcinoma de intestino humano. O material foi coletado no período entre 1998 e 2003 no Hospital Universitário de Santa Maria, RS, Brasil. Como controle, foram utilizadas amostras fígado humano para MMP-9 e células deciduais normais para MMP2. Foi realizada a técnica de imunohistoquímica e sistema de amplifica- ção por polímero não biotinilado Novolink (Novocastra, New Castle Upon Tyne, Inglaterra) para avaliar as metaloproteinases 2 e 9, por percentual de células neoplásicas coradas, semi-quantitativamente, considerando-se expressão citoplasmática. Resultados: Realizada análise estatística através do teste exato de Fisher, com nível de significância de 0,05, os testes não evidenciaram associação significativa para nenhuma das aplicações feitas. Conclusões: Os achados sugerem que não existe associação entre a expressão das metaloproteinases 2 e 9 com as variáveis deste estudo, sendo necessário estudos futuros (AU)

Introduction: Colorectal carcinomas are the third most common type of malignant neoplasia and the third leading cancer-related cause of death worldwide. Aims: This study aimed at determining if there is an association between staging, vascular invasion, tumor size, Duke and Astler-Coller classifications, tumor differentiation grade, and metalloproteinase 2 and 9 in intestinal adenocarcinoma. Methods: A retrospective study was carried out of 40 paraffin blocks containing samples from surgical resections of intestinal tissue previously diagnosed as colon adenocarcinoma. The material was collected from 1998 to 2003 in the School Hospital of Santa Maria, RS, Brazil. Human liver extracts for MMP9 and normal decidual cells for MMP2 were used as controls. Immunohistochemistry and non-tinylated polymer amplification system Novolink (Novocastra, New Castle Upon Tyne, England) were performed to evaluate metalloproteinases 2 and 9 by percentage of stained cytoplasmic neoplastic cells, semi-quantitatively, and considering the cytoplasmic expression. Results: In the statistical analysis, the Fisher's exact test was used with a level of significance of 0.05, which did not show any significant association for any of the applications done. Conclusions: The findings show that there is no association of metalloproteinases 2 and 9 expression with the variables studied here (AU)

Avaliação da expressão da caspase-3 no adenocarcinoma colorretal através de um sistema computadorizado de análise de imagem / Evaluation of caspase-3 markers expression in colorectal adenocarcinoma by a computadorized image analysis system

RACIONAL: O câncer colorretal é uma das causas mais comum de mortes por neoplasias malignas no mundo ocidental, sendo que o adenocarcinoma colorretal representa mais de 90 por cento dos tumores malignos que acometem o intestino grosso. A cisteíno-protease caspase-3, uma das mais participantes enzimas do processo apoptótico, pode ser utilizado como marcador tumoral. OBJETIVO: Identificar e quantificar a expressão da caspase-3 em amostras de adenocarcinomas colorretais através de um sistema computadorizado de análise de imagem, bem como correlacionar a expressão deste marcador. MÉTODOS: Foi estudada por imunoistoquímica a proteína caspase-3 em 55 amostras de adenocarcinoma colorretal parafinizadas. Para analisar os tecidos imunomarcados, o sistema SAMBA 4000 foi utilizado. Avaliaram-se as lâminas com o programa IMMUNO 4.00 através da leitura de 8 a 10 campos, determinando-se dois parâmetros: índice de marcagem (LI, Labeling Index) que descreve a porcentagem de área tecidual especificamente marcada pela prova imunoistoquímica, e a densidade óptica média (MOD, Mean Optical Density), a qual representa a intensidade de coloração do tecido medida por pixels positivos. Para a análise estatística, utilizou-se o teste t-Student e o coeficiente de Pearson para verificar a existência de correlação entre índice de marcagem e a densidade óptica média do mesmo marcador. RESULTADOS: Todas as amostras estudadas apresentaram positividade à caspase-3 (LI médio: 85,24 por cento ± 8,71 por cento de células positivas). Houve correlação estatisticamente significante entre o índice de marcagem e a densidade óptica média deste marcador. CONCLUSÃO: A intensidade da expressão de caspase-3 e o número de células expressando este marcador correlacionam-se positiva e significantemente.

BACKGROUND: Colorectal cancer is one of the most common causes of death for malignant neoplasm in the Occidental World, corresponding colorectal adenocarcinoma to represent more than 90 percent of malignant tumors in large bowel. Caspase-3 cysteine protease, one of the most important enzymes related in the apoptosis process, may be used as a tumor marker. AIM: To identify and quantify Caspase-3 expression in colorectal adenocarcinoma samples and to correlate the marker expression using an image computerized analysis system. METHOD: Caspase-3 expressions were studied by immunohistochemistry in 55 colorectal adenocarcinoma sampled-tissues, prepared in paraffin blocks. SAMBA 4000 System was used for the immunostained tissues analysis. Microscopic evaluation was performed by the IMUNNO 4.00 software, which analyzed 8 to 10 fields in order to determine two parameters: Labeling Index (LI), indicating the percentage of cells specifically stained by immunohistochemistry, and Mean Optical Density (MOD), representing enhance intensity, measured by positive pixels. For the statistical analysis, t-Student test and Pearson coefficient verified the possible correlation concerning LI and MOD. RESULTS: All samples presented positivity for Caspase-3 expression (LI 85,24 percent ± 8,71 percent positive cells). Caspase-3 expression revealed statistically significance correlation between LI and MOD (P = 0,013). CONCLUSIONS: The intensity of Caspase-3 expression and the number of positive cells expressing such a marker showed a positive and significant correlation.