Underexpression of LATS1 TSG in colorectal cancer is associated with promoter hypermethylation.

AIM: To investigate large tumor suppressor 1 (LATS1) expression, promoter hypermethylation, and microsatellite instability in colorectal cancer (CRC). METHODS: RNA was isolated from tumor tissue of 142 CRC patients and 40 colon mucosal biopsies of healthy controls. After reverse transcription, quantitative polymerase chain reaction (PCR) was performed, and LATS1 expression was normalized to expression of the ACTB and RPL32 housekeeping genes. To analyze hypermethylation, genomic DNA was isolated from 44 tumor CRC biopsies, and methylation-specific PCR was performed. Microsatellite instability (MSI) status was checked with PCR using BAT26, BAT25, and BAT40 markers in the genomic DNA of 84 CRC patients, followed by denaturing gel electrophoresis. RESULTS: Decreased LATS1 expression was found in 127/142 (89.4%) CRC cases with the average ratio of the LATS1 level 10.33 ± 32.64 in CRC patients vs 32.85 ± 33.56 in healthy controls. The lowest expression was found in Dukes' B stage tumors and G1 (well-differentiated) cells. Hypermethylation of the LATS1 promoter was present in 25/44 (57%) CRC cases analyzed. LATS1 promoter hypermethylation was strongly associated with decreased gene expression; methylated cases showed 162× lower expression of LATS1 than unmethylated cases. Although high-grade MSI (mutation in all three markers) was found in 14/84 (17%) cases and low-grade MSI (mutation in 1-2 markers) was found in 30/84 (36%) cases, we found no association with LATS1 expression. CONCLUSION: Decreased expression of LATS1 in CRC was associated with promoter hypermethylation, but not MSI status. Such reduced expression may promote progression of CRC.

The mRNA level of the transforming growth factor ß1 gene, but not the amount of the gene product, can be considered as a potential prognostic parameter in inflammatory bowel diseases in children.

PURPOSE: Transforming growth factor ß1 (TGF-ß1) plays a role in cell proliferation and differentiation, and it can modulate immune response. In this work, we asked whether levels of either TGF-ß1 or mRNA of the corresponding gene in plasma or tissue can be useful in diagnosing and/or monitoring of the clinical course of inflammatory bowel diseases (IBD). METHODS: The study group consisted of 104 pediatric patients with IBD: 36 with Crohn's disease (CD) and 68 with ulcerative colitis (UC); 42 children represented the control group. TGF-ß1 levels in plasma and intestinal mucosa were estimated by ELISA and immunohistochemistry (IHC), respectively. Levels of TGF-ß1 mRNA were determined by reverse transcription and real-time PCR. RESULTS: In patients with IBD, and in subgroups with CD and UC, no significant differences in the TGF-ß1 level in plasma and tissue were found relative to the control group. These variables were not dependent on the stage of the disease, its activity or severity of endoscopic and histopathological findings. TGF-ß1 mRNA levels were significantly higher in tissue samples withdrawn during the relapse of the disease than in those taken during the remission or in the control group. However, no correlation between TGF-ß1 plasma levels and TGF-ß1 mRNA amount in the intestinal mucosa was observed. CONCLUSIONS: The TGF-ß1 mRNA level, but not the amount of the gene product, was significantly increased in the pathologically changed tissue during the relapse of IBD. We suggest that this parameter might be considered as a potential prognostic value when assessing IBD in children.

[Role of transforming growth factor beta1 (TGF-beta1) in the pathogenesis and clinical course of chronic hepatitis in children]. / Rola transformujacego czynnika wzrostu beta1 (TGF-beta1) w etiopatogenezie i przebiegu klinicznym przewleklych zapalen watroby u dzieci.

UNLABELLED: Transforming growth factor-beta1 (TGF-beta1) is known to play a key role in processes of cell proliferation and differentiation. It also plays an important role in modulation of the immune response. Various diseases may arise both from excessive and insufficient activity of this cytokine. THE AIM OF THE STUDY was to evaluate the role of TGF-beta1 in the pathogenesis of chronic hepatitis (Ch.h.) and to assess whether TGF-beta1 level in plasma or its tissue expression can be useful in diagnosing and monitoring of clinical course of this disease. PATIENTS AND METHODS: Twenty-one children with chronic hepatitis were included in the study and 42 healthy children constituted the control group. Liver function tests and TGF-beta1 plasma levels measured by ELISA method were evaluated in both groups of patients. In liver tissue obtained by needle biopsy, the histopathological grading and staging of hepatitis was evaluated, TGF-beta1 protein was assessed by immunohistochemical methods and TGF-beta1 gene expression was measured by reverse transcription and real-time polymerase chain reaction. RESULTS: In chronic hepatitis group of patients the plasma TGF-beta1 level did not differ from the control group and did not correlate with grading and staging of the liver tissue while positive correlation was observed with gamma-glutamyl transpeptidase activity in the serum. There was no correlation between tissue TGF-beta1 expression and TGF-beta1 plasma level and staging or grading in liver tissue. TGF-beta1 gene expression correlated positively with ESR and ALAT activity but no correlation with TGF-beta1 plasma level, TGF-beta1 gene or protein expression, grading or staging in liver tissue were observed. CONCLUSION: 1. In children with chronic hepatitis, TGF-beta1 plasma level is not related to grading or staging in the liver tissue. This finding may be due to low level of fibrosis observed in the studied children. 2. It appears that local expression of TGF-beta1 in liver tissue should not be used as a sole marker in differentiating and monitoring the course of chronic hepatitis. 3. In children with chronic hepatitis assessment of liver TGF-beta1 gene expression is not helpful in the evaluation of pathological changes in liver tissue. 4. Due to the relatively low number of patients in the analysed groups it seems advisable to perform similar complex studies in larger groups of children with chronic hepatitis.

High serum interleukin-18 concentrations in patients with coronary artery disease and type 2 diabetes mellitus.

AIMS: The aim of our study was to analyse the serum level of interleukin 18 (IL-18) in coronary artery disease (CAD) patients with type 2 diabetes mellitus (DM), and to relate this to clinical findings. METHODS: The IL-18 level was measured by ELISA in serum samples from 130 CAD patients prior to their first, elective, coronary artery bypass surgery. Forty-three of them had been diabetic for several years. A control group consisted of 31 healthy people matched according to age, BMI, lipid and smoking status. RESULTS: The CAD patients with DM were similar to the non-diabetic CAD patients with respect to age, BMI, grade of heart failure, ejection fraction. There were no differences in the duration of CAD, history of myocardial infarction and PTCA or instability of angina. The serum level of IL-18 was higher in the CAD patients than in the control group. The CAD patients with DM had a higher concentration of IL-18 compared to the non-diabetic CAD group. The diabetic patients with triple-vessel disease were characterized by a higher concentration of IL-18 than the non-diabetic patients with the same grade of CAD. Smoking affected the IL-18 concentration, particularly in the diabetic patients. CONCLUSION: Type 2 DM predisposes patients, especially those with multi-vessel CAD who were smokers, to a higher serum level of IL-18, which may help explain their vulnerability to fatal, secondary cardiovascular events. These patients should be in the first line for stringent, secondary cardiovascular prevention.