الفهرس 
AcknowledgementOnly 14 pages are availabe for public view
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Abstract
Introduction: Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the second most common cause of cancer related death. Currently available screening tests to detect early stage of HCC are combined α-fetoprotein (AFP) analysis and ultrasound. However, screening for early detection of HCC has become quite common in clinical practice, its effectiveness remains controversial Several genetic and epigenetic alterations were shown to be related with hepatocarcinogenesis, including frequent promoter hyper methylation of several tumor suppressor genes such as Ras association domain family 1 isoform A (RASSF1A), E Cadherin gene (Cdh1) and RUNX3. The aim of work : Evaluation of the methylation of multiple tumor suppressor genes in the serum of patients with HCV related liver cirrhosis as well as Hepatocellular carcinoma as possible serum biomarkers for early detection of HCV related hepatocellular carcinoma.Research Plan: The study included 53 Egyptian healthy control volunteer, 18 chronic hepatitis C patients, 189 HCV related liver cirrhosis (LC) patients and193 HCV related HCC patients attending the clinics of Tropical Medicine Department. Laboratory investigations were done in the molecular genetic unit of endemic hepatogastroenterology and infectious diseases (MGUEHGID). All participants were subjected To : • Full history taking and Clinical examination. • Radiological investigations : Abdominal ultrasonography U/S and spiral CT for detection of hepatic focal lesions, Molecular study : detection of hyper methylated RASSF1A, e-cadherin and RUNX3 gene by real time methylation specific PCR. Results : 1- Significant hypermethylation of RASSF1A, e-cadherin and RUNX3 genes in HCV associated liver cirrhosis. 2- Significant hypermethylation of RASSF1A, e-cadherin and RUNX3 genes in HCV associated HCC. 3- Significant increase in hypermethylated RASSF1A, e-cadherin and RUNX3 genes in HCV associated HCC than liver cirrhosis and apparently healthy control. 4- Performance of the hyper methylated RASSF1A, E Cadherin and RUNX 3 genes as new biomarkers for prediction of HCV related HCC from peripheral blood DNA by MSP revealed that at cut off levels of >0.389 , >0.14 and >0.23 respectively, the sensitivities were 46.36%,83.96% and 76.19% respectively and the specificities were 86.7% ,59.46% and 58.6% respectively.5-No significant correlations were found between AFP level and methylation level of candidate genes in LC and HCC groups. 6- No significant association was found between methylation level of candidate genes and clinical data within the studied groups. 7- No significant association was found between methylation level of candidate genes and biochemical data within the studied groups. 8- After Logistic regression analysis for prediction of HCC within LC cases, using age, gender, smoking, AFP, candidate genes as covariates, RASSF1a and E Cadherin genes hypermethylation were considered as predictors of HCC within LC cases in uni and multivariable analyses. 9- After logistic regression analysis for prediction of HCC within LC cases with low AFP. RASSF1a, E Cadherin and RUNX3 genes hypermethylation were associated with risk of HCC within LC cases in univariable analysis. 10- Also after multivariable regression analysis for prediction of HCC within LC cases with low AFP. E Cadherin gene hypermethylation was considered as the only independent risk factor for prediction of HCC with low AFP. 11- Performance of the combined hyper methylated RASSF1A, E Cadherin and RUNX 3 genes at cut off levels of >0.389 , >0.14 and >0.23 respectively, showed significantly higher AUCs = 0.858, Sensitivity= 71.6% and Specificity= 84.9% revealing that they could be as new biomarkers from peripheral blood DNA by MSP that were conducted for discrimination between HCV related HCC and LC cases. Conclusion : • Hypermethylation of RASSFIA, RUNX3 and E Cadherin genes in peripheral blood could be a noninvasive marker for diagnosis of hepatocellular carcinoma (HCC). • Hypermethylation of RASSFIA and E-Cadherin genes in peripheral blood could be predictor risk factor of HCC in HCV associated liver cirrhosis patients. •E Cadherin gene hypermethylation was considered as the only independent risk factor for prediction of HCC with low AFP. • The presence of hypermethylated RASSF1A, RUNX3, and E Cadherin in serum may be useful biomarkers for HCC early prediction in patients of LC with HCV. •The prognostic and therapeutic values of hypermethylation of RASSFIA, RUNX3 and E Cadherin genes in peripheral blood of HCC patients need further research.