Autophagy-related gene expression in colorectal cancer patients.

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.

The profile of melatonin receptors gene expression and genes associated with their activity in colorectal cancer: a preliminary report.

The antiproliferative and immunomodulatory effects of melatonin (MLT) have been demonstrated in a variety of neoplasms including colorectal cancer (CRC). In humans and other mammals, MLT acts on target tissues through membrane and retinoid nuclear receptors. The aim of this study was to evaluate transcription activity of melatonin receptors and genes associated with regulation of their activity in colorectal adenocarcinoma tissues in relation to clinical stage of cancer. A total of 24 pairs of surgically removed tumoral and healthy (marginal) tissue samples from colorectal cancer patients at clinical stages I-II and III-IV were collected. As an additional control, twenty normal samples were tak¬en from people whose large intestine tissues were reported as non-tumoral after colonoscopy. Expression of mRNA genes was studied by microarray HG-U133A analysis. The analysis of gene expression profile was performed using commercially available oligonucleotide microarrays of HG-U133A. High increase of MT1 mRNA expression levels in all cancerous samples vs non-cancerous tissues was observed. The MT2 mRNA expression levels increased slightly in marginal and malignant samples. Among the genes participating in the cascade of signal transfer in cells activated by MLT via melatonin receptors, we found encoding genes (GNA11, OXTR, TPH1) only for differentiating stage III - IV of CRC. Monitoring the expression levels of genes that are related to melatonin receptors may offer a strategy to anticipate tumour development and estimate the molecular changes that occur during carcinogenesis. The mechanism behind this association needs further elucidation.

Expression of ADAM28 and IGFBP-3 genes in patients with colorectal cancer - a preliminary report.

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.

Statistical analysis of differential gene expression in colorectal cancer using CLEAR-test.

CLEAR test provides a novel method of analysis by combining inference for differential expression and variability. Frozen tumor specimens from 14 (3 coded Stage I, 5 Stage II, 2 Stage III and 4 Stage IV) colon cancer patients were obtained. Archived primary tumor samples were collected at the time of surgery and normal colon mucosae (controls specimens) were also collected. The studied transcriptomes were clustered using hierarchical agglomeration with Ward's method and Tchebychev distance. The separable groups of transcriptomes were classified as high clinical stage of adenocarcinoma (HCS; stages II-IV), low clinical stage of adenocarcinoma (LCS; stages I and 3 controls), and two normal colon mucosae (controls N1 and N2). The results of the CLEAR-test algorithm in normal colon specimens and adenocarcinoma specimens with low and high clinical stage showed 50 most and 50 least significant genes. The list of differential genes (p<0.01) in normal colon specimens and adenocarcinoma specimens with low and high clinical stage presented 58 genes.

Genetic disregulation of gene coding tumor necrosis factor alpha receptors (TNFalpha Rs) in colorectal cancer cells.

The expression of TNF ligand by malignant cells might be a mechanism for tumour immune escape. Genetic disregulation of gene coding TNF receptors was observed in neoplastic disease by an increased number of receptors on tumour cells and ligand-receptor activity. It might cause tumour proliferation and metastatic potential. Structure of TNF receptors influences TNF activity in vivo and structure of TNF R2 gene may suggest post-transcription modification based on alternative splicing. The aim of the study was to analyse the expression of gene coding TNF receptors R2 and R2/R7 (without exon 7) by estimation of mRNA expression of colorectal cancer cells in comparison with surrounding tissue free from neoplastic infiltration and searched for differently spliced TNFalphaR2/R7 isoforms. The study included fifty four patients with histopathologically confirmed adenocarcinoma (Stage III according to the AJC TNM Classification). Tissue samples removed from the tumour region were obtained from colorectal cancer patients undergoing surgical treatment. The samples were divided into two parts. The first one--was routinely examined histopathologically, the second one--was used for RNA extraction and the number of TNF and its receptors mRNA copies were subsequently quantified. The TNF and TNFRII genes expression were estimated based on the number of mRNA copies on 1 microg total RNA. The presence of TNFR2 and TNFR2/R7 isoforms in tumour, normal and metastatic cells was observed. The highest number of mRNA TNF copies and over expressed TNF genes were investigated and significantly noticed in metastatic cells (lymph nodes). The decreased number of TNFR2/R7 mRNA copies in metastatic lymph nodes secondarily influenced the decreased TNF soluble receptors' concentration. In conclusion, the genetic disregulation observed in neoplastic disease usually concerns dysfunction of cytokines receptor genes.

On the holistic approach in cancer biology: tumor necrosis factor, colon cancer cells, chaos theory and complexity.

TNFalpha plays a role in the pathogenesis of septic shock, inflammatory diseases, autoimmune diseases, graft rejection reaction, acute, and chronic respiratory inefficiency among others. Its activity depends on the type of target cells and different regulating factors, but the effect of biological activity is conditioned by specific receptors such as p55 (type I, TNF R55) and p75 (type II, TNF R75). The aim of the study was to answer the following questions: 1) Is it possible to apply elements of non-linear dynamics to assess the level of expression of TNF, TNFRI, TNFRII genes in tumor cells, pathologically unchanged tissue and metastatically changed lymph nodes? 2) Is theoretically anticipated variability of cytokine and its receptors in colorectal carcinoma cells and the immediate vicinity justified in the developed mathematical model? The research material--specimens taken from tumor, unchanged tissue and metastatic lymph nodes--were histopathologically and molecularly analysed. Results of the molecular research were used to develop a mathematical model using the basic studies on the theory of chaos and biological system modelling.

Circadian fluctuations of melatonin, tumor necrosis factor-alpha and its soluble receptors in the circulation of patients with advanced gastrointestinal cancer.

The objective of the study was to investigate the dynamic changes of melatonin (MLT), tumor necrosis factor alpha (TNFalpha), soluble TNFalpha receptors ( type I and type II) in serum of advanced cancer patients during 24 hours. The examined group consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Blood samples were collected 6 times a day (8 a.m., 2 p.m., 6 p.m., 10 p.m., 2 a.m., and again 8 a.m.) as well as in healthy controls. Serum levels of TNFalpha and both its receptors were measured using ELISA type and the radioimmunoassay method was used to assess MLT levels. The circadian rhythm of MLT was altered because MLT reached its peak level at 8.50 a.m. with 5 hours delay in respect to average peak time in healthy humans. The presence of circadian rhythm of TNFalpha was proved (acrophase-1.40 a.m.), and no diurnal rhythm of soluble TNF receptors was observed. The concentration of soluble type I (p-55) receptor was distinctly lower than soluble type II (p-75). The peak of soluble type I receptor value appeared at 10.00 p.m. while the type II receptor reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations, the shapes of both curves remained inversely proportional. The present results may suggest the presence of complex self-regulation mechanisms between the neuroendocrine system and the cytokine network in advanced gastrointestinal cancer patients.

Genetic disregulation of gene coding tumor necrosis factor alpha receptors (TNF alpha Rs) in follicular thyroid cancer--preliminary report.

UNLABELLED: TNF alpha receptors participate in programmed cell death. TNF R2 efficiently assists TNF R7 effects by ligand passing. Structure of TNF alpha receptors influences TNF activity in vivo and the structure of TNF R2 gene may suggest post-transcription modification based on alternative splicing. The aim of the study was to analyse the expression of gene coding TNF alpha receptors R2 and R7 by estimation of mRNA expression of differentiated thyroid carcinomas in comparison to surrounding tissue free from neoplastic infiltration and search for differently spliced TNF alpha R2/R7 isophorms. The study included seven patients with histopathologically confirmed follicular thyroid cancer. Tissue samples removed from tumor region were obtained from the follicular thyroid cancer patients undergoing surgical treatment. The samples were divided into two parts. The first one was routinely examined histopathologically, the second was used for RNA extraction and the number of TNF and its receptors mRNA copies were subsequently quantified. RESULTS: 1) The presence of TNF alpha expression was observed in all examined samples, in contrast to TNF R1 expression; 2) The high level of TNF alpha expression was noted both for typical and sought TNF R2/R7 isoforms and 3) A considerable number of samples displayed higher levels of TNF R2 isoforms than TNF R2/R7 mRNA expression. Genetic disregulation observed in neoplastic disease usually concerns dysfunction of cytokines receptor genes.

Dynamics of circadian fluctuations in serum concentration of cortisol and TNF-alpha soluble receptors in gastrointestinal cancer patients.

The objective of the study was to investigate the dynamic changes of cortisol and soluble TNF-alpha p-55 and p-75 receptors in serum of advanced cancer patients during 24 h. The examined group consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Serum levels of cortisol, TNF-alpha and both receptors in cancer patients were measured using ELISA type kits 6 times a day (8.00 a.m., 2.00 p.m., 6.00 p.m., 10.00 p.m., 2.00 a.m. and again 8.00 a.m.) as well as in healthy controls. The levels of cortisol, TNF-alpha and its soluble receptors were substantially increased in examined group and displayed statistically significant circadian fluctuations. Cortisol peak was found typically at 8.08 a.m., the minimum value appeared at 6 p.m. The presence of circadian rhythm of the cytokine was proved (acrophase -00.36 a.m.), however no diurnal rhythm of soluble TNF receptors was observed. The concentration of p-55 receptor was distinctly lower then p-75. The peak p-55 value appeared at 10.00 p. m. while the p-75 reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations the shapes of both curves remained in inversely proportional manner to each other. The present results may suggest the presence of complex self-regulation mechanisms in advanced gastrointestinal cancer patients. There was no correlation observed between cortisol and TNF-alpha soluble receptor concentration.

Dynamics of diurnal changes in serum concentration of TNF-alpha soluble receptors in gastrointestinal cancer patients.

The objective of the study was to investigate the dynamic changes in soluble TNFalpha p-55 and p-75 receptors in serum of advanced cancer patients during 24 hours. The group examined consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Serum levels of the cytokine and both receptors in cancer patients were measured using ELISA type kits 6 times a day (8.00 a.m., 2.00 p.m., 6.00 p.m., 10.00 p.m., 2.00 a.m. and again 8.00 a.m.) as well as in healthy controls. The levels of TNFalpha and its soluble receptors were substantially increased in the examined group and displayed statistically significant circadian fluctuations. The presence of circadian rhythm of the cytokine was proved (acrophase - 00.36 a.m.), however no diurnal rhythm of soluble TNF receptors was observed. The concentration of p-55 receptor was distinctly lower then p-75. The peak p-55 value appeared at 10.00 p.m. while the p-75 reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations the shapes of both curves remained inversely proportional. The present results may suggest the presence of complex self-regulation mechanisms in advanced gastrointestinal cancer patients.

[Soluble tumor necrosis factor receptors--their properties and role in diagnosis and therapy of acute and chronic diseases]. / Rozpuszczalne receptory czynnika martwicy nowotworów--ich wlasciwosci i znaczenie w diagnostyce i terapii schorzen o przebiegu ostrym i przewleklym.

The review presents the properties and significance of soluble forms of tumor necrosis factor-alpha receptors in physiology and pathology of human organism. Special attention was turned to the possibility of practical application of those forms of the receptors in the therapy of diseases caused by excessive activity of TNF-alpha. The significance of the soluble receptors as prognostic factors of disease progression and outcome was stressed.

The estimation of endogenous tumor necrosis factor alpha and cortisol levels in serum in advanced neoplasm.

The immune reaction mediators--cytokines may play an important role in central nervous system function and influence the neuroendocrine system. The aim of the study was to assess the hypothesis of a possible correlation between endogenous TNF-alpha and glucocorticoids secretion in advanced neoplasm. Thirty patients (17 males and 13 females) with advanced neoplasm were enrolled in the study. The levels of TNF-alpha were measured with ELISA type test and cortisol levels were estimated by RIA method. The blood samples were collected 6 times during 24-hours for TNF and 6 times a day for cortisol. Twenty healthy volunteers were studied in parallel. Both TNF-alpha and cortisol levels revealed the presence of circadian rhythm in examined patients. The levels of TNF-alpha were very high and achieved the peak value about midnight (acrophase-01.40). Cortisol peak was found typically at 08.08. The fluctuations of examined molecules showed regular patterns. TNF-alpha levels markedly decreased together with the increase of cortisol and rose when cortisol concentration in serum was low. Although the correlation was not confirmed by Spearman correlation rank the shapes displayed by both curves may suggest the presence of neuroendocrine feed-back loop between corticosteroids and TNF-alpha in advanced cancer.

Circadian rhythmometry of serum endogenous tumor necrosis factor-alpha in patients with colorectal cancer metastases.

The objective of this study was to test the hypothesis of a diurnal fluctuation of circulating levels of endogenous tumor necrosis factor-alpha (TNF-alpha) in patients with colorectal cancer metastases. Serum levels of TNF-alpha were measured 6 times a day in 27 colorectal cancer patients as well as in healthy controls. The levels of TNF-alpha in serum from healthy controls were close to the detection limit of the method or were not detectable at all. Serum TNF-alpha levels in advanced cancer patients were substantially increased. Peak values appeared at 2 a.m. In contrast, the levels were low in the afternoon (2 p.m). The presence of a circadian rhythm was found with the acrophase at 0 hour 40 min.

Is there a correlation between tumor necrosis factor alpha and soluble p-55 receptor plasma levels in advanced gastrointestinal cancer patients?

UNLABELLED: Studies have shown an increased spontaneous TNF alpha production, and the 24--rhythmicity of TNF blood concentration in patients with advanced cancer. The present study investigates whether diurnal rhythmicity of endogenous TNF alpha is associated with the induction of circulating sTNF Rp55 in advanced gastrointestinal neoplasm. The levels of endogenous TNF alpha and sTNF Rp55 in serum were measured at 8 a.m., 2 p.m., 6 p.m., 10 p.m., 2 a.m., and again at 8 a.m. RESULTS: There is circadian rhythm in the secretion of endogenous TNF alpha in patients with advanced gastrointestinal cancer, however no diurnal rhythm of sTNF Rp55 was observed. Since it has been suggested that human tumors susceptible to TNF alpha may escape destruction by secreting or inducing the secretion of sTNF Rp55 to block the effects of TNF alpha, the possibility that the observed fluctuations could also reflect the complexation of a proportion of the receptor molecules with their ligand should be considered.

Human recombinant tumor necrosis factor alpha in combined therapy of advanced pancreatic cancer.

Preliminary results of a combined therapy consisting of interferon-alpha, human recombinant tumor necrosis factor-alpha and 5-FU in patients with advanced pancreatic cancer are presented. Each patient underwent surgical treatment, except one case in which partial resection was done. In all other patients only palliative operation was performed because of the advanced stage of the disease. The patients have been followed up for 4-19 months and all of them are still alive. Complete remission was observed in one patient; survival time 13 months. In two patients partial remission was achieved, survival time 10 and 6.5 months, respectively. The general condition of other patients was good and stable during the follow up.