الفهرس 
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Abstract
The main aim of the current study is to investigate HCV core polymorphism and its potential association with HCC in a comparative genetic analysis study merely focused on HCV core region in HCV chronically infected Egyptian patients. Also the study aimed to reveal the following; possible association of HCV viral load with HCC, association of hepatic fibrosis with HCV core polymorphism in addition to determination of HCV genotypes and subtypes by direct sequencing of the core gene of HCV, and finally to determine predictive risk factors for HCC development.
This study was carried out on sixty HCV seropositive RNA positive subjects divided into two groups:
• Cases included 30 chronic HCV patients with hepatocellular carcinoma.
• Control included 30 chronic HCV patients free from hepatocellular carcinoma.
Sera from all patients were tested by the following molecular investigations:
• RNA extraction was performed using Qiagen QIAamp viral RNA mini spin protocol.
• Detection of HCV RNA viral load using Real time PCR using Artus HCV QS-RGQ PCR Kit.
• Amplification of core region of HCV genome by conventional nested PCR.
• Direct sequencing of HCV core gene using Sanger sequencing technique.
• Determination of HCV genotypes and subgenotypes after sequencing the core region using HCV BLAST software.
• Comparison of the results with reference strains to uncover the presence of mutations using BioEdit software.
The most striking result of our study is that HCC core genes do have characteristic mutations rather than those of HCV core genes in the control group. In another words, significant association of HCV core polymorphism with HCC was reported in HCV GT-4 Egyptian patients. Moreover, significant association of HCV core aa substitutions at position 71 (core-p71) was reported with HCC, but not at position 70 nor position 91 which were frequently reported by other investigators on HCV GT-1a.
Significant association between advanced age and HCC was reported in the current study as the majority of cases (HCV patients with HCC) were above the age of 50 (70%), while the majority of control group (HCV patients without HCC) were below that age (70%).
We demonstrated that the majority of patients in both study groups were males (80% in cases and 70% in control). The male to female ratio was 4:1 in the group of cases and 3:1 in the control group.
The current study also demonstrated that past history of the patients that may predispose for acquisition of HCV infection is more significantly reported with dental procedures while is least frequently reported with blood transfusion.
In the present study laboratory liver related investigations were used to assess the status of liver disease among both study groups. Our results revealed significant difference between the two groups regarding liver transaminases (ALT and AST), AST/ALT ratio, GGT, total bilirubin, alkaline phosphatase (ALP), and alpha fetoprotein (AFP) level.
The results revealed that all patients (100 %) from both study groups belonged to genotype 4. In cases, 73.3% were of subgenotype 4a, followed by 13.3% subgenotype 4l then subgenotype 4n and 4o in equal percentage which is 6.7%. Using the same technique for subgenotyping HCV amplicons from control group, 90% were of subgenotype 4a followed by 10 % subgenotype 4l, 4n and 4o in equal percentage which is 3.3%. Although HCV subgenotype 4l was reported in HCC cases 4 times more than in the control group, that difference was not statistically significant. Significant association between other HCV subgenotypes and HCC do not exist.
In our study we revealed that there is no significant difference between HCV viral load in both HCV patients with or without HCC.
We demonstrated a significant association between advanced hepatic fibrosis with HCC development and also HCV core aminoacid heterogeneity with advanced degree of hepatic fibrosis in both study groups.
The results of our study of multivariate analysis of factors predicting HCC development among patients with HCV-GT4 demonstrated that AST, ALT, AST/ALT ratio, GGT, ALP, AFP, HCV core aminoacid mutations and hepatic fibrosis score; all were found to be important predictors for having HCC among HCV patients. Furthermore, Advanced hepatic fibrosis, AFP and HCV core aa mutations were reported as the most important predictors for HCC risk.