RESUMO
There is evidence that autophagy can play a dual role in tumor cells as a tumor suppressor, and a process involved in tumor cell survival. The aim of this work was to assess the expression of the genes engaged in the autophagy process in biopsies taken from the colon, confirmed as adenocarcinoma, and normal tissue and to relate them to the clinical stage of the tumor. A total of 20 pairs of surgically removed tumors and healthy (marginal) tissue samples from colorectal cancer patients at clinical stages (CS) I-IV were analyzed. Gene expression profile analysis was performed using HG-U133A microarrays. Differentially expressed genes were identified, using the PL-Grid Infrastructure. Only for CSI, there were two specific genes: FOXO1 and BNIP1; further in CSII LAMP2, MET and BCL2L, in CSIII HIF1A and 2 ID mRNAs for HGF and 18 genes were specific for CSIV in comparison to controls. PINK1 is the only gene that differentiates all transcriptome groups from controls. Furthermore, examination of the expression of genes associated with the autophagy process may allow for better knowledge and understanding of the processes occurring during the development of colon cancer. The presented genes may be used as prognostic markers of clinical stages of colorectal cancer, contributing to the development of new lines of therapy focused on reducing metastasis of the primary tumor.
Assuntos
Adenocarcinoma/genética , Autofagia/genética , Colo/metabolismo , Neoplasias Colorretais/genética , Regulação Neoplásica da Expressão Gênica , Transcriptoma , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Colo/patologia , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Proteína Forkhead Box O1/genética , Proteína Forkhead Box O1/metabolismo , Perfilação da Expressão Gênica , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Proteína 2 de Membrana Associada ao Lisossomo/genética , Proteína 2 de Membrana Associada ao Lisossomo/metabolismo , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/metabolismoRESUMO
Phosphatase and Tensin Homolog deleted on chromosome 10 (PTEN) gene is one of the most important tumor suppressor genes which is involved in the regulation of many signaling cascades (AKT/PKB and MAPK). Subtle changes in its activity lead to cancer susceptibility or aggressive tumor behaviour. Despite the diversity of mechanisms leading to PTEN inactivation, it is frequently associated with a decreased or complete loss of protein expression. About 20% decrease in PTEN expression could lead to the development of cancer. There have been no objective, quantitative methods of PTEN expression assessment that allow to measure the subtle variations of the protein concentration in a tissue-contextual manner. A new quantitative algorithm of immunostaining evaluation based on combination of color deconvolution and relative chromogen signal intensity was used in the study. The proposed algorithm was implemented in the popular ImageJ image analysis software and positively verified in cancer cell lines and tissue models as well as in the tissue samples of colorectal cancer (CRC) patients. The proposed quantitative method of PTEN expression assessment creates an alternative to currently available subjective methods and forms the basis for inter-case and inter-tissue comparisons. Using the algorithm it would be possible to identify three groups of patients with advanced colorectal cancer which could significantly differ in the overall survival. The research should be continued.
Assuntos
Neoplasias/enzimologia , PTEN Fosfo-Hidrolase/metabolismo , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Linhagem Celular Tumoral , Compostos Cromogênicos/metabolismo , Epitélio/metabolismo , Epitélio/patologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Neoplasias/patologiaRESUMO
Adamalisynes (ADAMs) play an important role in inter-membrane interactions, cell adhesion and fusion processes and protein shedding from the cell surface. Many reports indicate that members of the ADAMs family are overexpressed in human cancer. The aim of the present study was to evaluate ADAM28 and Insulin Like Growth Factor Binding Protein-3 (IGFBP-3)) gene expression in colorectal carcinoma tissues with regard to the overweight or obese status of the patients using an oligonucleotide microarray technique. Fresh tissue specimens were obtained from colorectal cancer patients during surgical treatment. Eighteen specimens from tumour and 18 normal tissue specimens from colorectal cancer patients at clinical stages III and IV were analysed. The examined patients were divided into two groups; those with BMI greater than or equal to 25 and those with normal BMI. The control group consisted of 18 specimens of non-neoplastic colon tissues, which were divided between overweight/obese and normal body weight patients. The gene transcriptional activity from the specimens was analysed using an oligonucleotide microarray technique. Microarrays and rinsing and marking solutions were prepared according to the procedure in the Gene Expression Analysis Technical Manual. The following conclusions were made: i) change of ADAM28 and IGFBP-3 genes expression are present in the normal tissue in overweight/obese patients with colorectal cancer only; ii) the observed molecular variability of ADAM28 and IGFBP-3 expression may be an initial process of cancer proliferation; iii) the histopathologically normal surgical margin in this group of patients was not equal to the molecular margin.
Assuntos
Proteínas ADAM/genética , Adenocarcinoma/genética , Neoplasias Colorretais/genética , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Sobrepeso/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Análise de Sequência com Séries de OligonucleotídeosRESUMO
Colorectal cancer is one of the most commonly diagnosed neoplasms still associated with relatively high mortality. Viral infections are often mentioned among the neoplasm transformation risk factors. Incidence of human papilloma virus (HPV), associated with high oncogenic risk, in the large intestine and the meaning of its presence in the colorectal carcinogenesis are still not clear. The aim of the study was to show a presence of HPV in specimens of adenomatous polyps and colorectal cancer using the Q-PCR method. Fifty patients (32 M/18W, mean age 62.8 years) were enrolled in the study, for whom tissue samples were obtained. Study material involved paraffin blocks derived from samples collected by flexible sigmoidoscopy from 10 polyps and 10 large intestine adenocarcinomas and 30 paraffin blocks with specimens of surgically removed large intestine adenocarcinomas. Presence of HPV genome was confirmed by quantitative PCR method using commercially available Abbott RealTime High Risk HPV test. The test is able to detect 14 most prevalent high oncogenic risk subtypes of human papilloma virus. Status of HPV DNA was successfully assessed in all 50 samples. No HPV DNA was discovered in any of the tested samples. Presence of high oncogenic risk HPV subtypes in large intestine adenoma and adenocarcinoma seems to be very rare, and its dominating role in the pathogenesis of colorectal cancer, even if possible, is unlikely.
