الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
epatocellular carcinoma (HCC) is the most common type of primary liver cancers with cirrhosis being the major risk factor for its development, which occurs usually due to HBV and HCV infection. Egypt has the highest prevalence of HCV worldwide and thus has rising rates of HCC representing a major health problem in Egypt, which highlights the urgent need for novel biomarkers for early diagnosis of HCC. HCC diagnosis depends on imaging techniques, serological tumor markers analysis and histopathological confirmation. Imaging techniques include US, CT& MRI however their sensitivities for detection of small HCC nodules might be low while the histopathological confirmation is an invasive technique and may cause tumor seeding. As regards serological tumor markers, currently the most widely used diagnostic marker is serum AFP, which has many limitations as it is not significantly elevated in many HCC patients particularly those in early stages which is potentially curable. Moreover, AFP could be elevated in cirrhotic patients or during exacerbations of chronic viral hepatitis without HCC. Due to these limitations, AFP has been excluded from HCC surveillance or diagnosis by the
H
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American Association for the Study of Liver Diseases guidelines (AASLD). So there is urgent need for new biomarkers that would be simple, minimally-invasive and reliable for the early diagnosis of HCC.
Exosomes contains proteins, lipids, RNA molecules, and cell-free circulating DNA. There is a great association between exosomes and hepatocellular carcinoma (HCC). Exosomes promote the progression of HCC by regulating the malignant behaviors of HCC cells including growth, migration, and invasion.
Due to previous stated informations, we used an integrative approach based on bioinformatics analysis together with clinical validation to provide great insights into the molecular mechanisms of HCC. To the best of our knowledge it is the first time, we assessed a competing endogenous network of Ras-related in brain 11 gene (RAB11A), which was selected according to public microarray databases, because this gene plays a major role in exosomal secretion. To confirm the expression of RAB11A gene (RAB11A mRNA) in HCC cases; we searched Gene Atlas database and protein Atlas database. Next, lncRNA-RP11-513I15.6, was identified, using a database of lncRNAs that act as ceRNAs (the lnCeDB database). This
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lncRNA acts as master regulator of the RAB11A by competing for miR1262 binding RAB11A gene
This study was done at Medical Biochemistry Department, Faculty of Medicine, Ain Shams University during the period from March 2015 – October 2016 and included 60 HCC patients 42 CHC patients and 18 normal volunteers.
The aim of the current study was to evaluate the clinical utility of serum levels of exosomal RAB11A-mRNA, miRNA-1262 and LncRNA-RP11513I15.6 expression as non-invasive biomarkers in diagnosis of HCC by quantitative Real Time -PCR and to correlate the expression of these markers to the various clinico-pathological parameters of the patients. This was done in an attempt to evaluate their role in HCC assessment and their use as potential selected diagnostic biomarkers. And to characterize the efficacy of Proton Pump Inhibitor PPI on liver function tests, liver histopathology, modulating exosomal production and exosomal RNA related to HCC using animal models.
The study included
120 subjects classified into three main groups:
Group1: 60 Hepatocellular carcinoma (HCC) cases with median age 60 years.
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group 2: 42 chronic hepatitis C (CHC) cases with median age 56.5 years.
group 3: 18 Healthy normal individuals with median age 53 years.
All studied subjects in this study were subjected to full clinical and radiological examination preceded by complete history taking and routine laboratory investigations including liver function tests as serum ALT, serum albumin, serum total bilirubin, serum AFP and INR.
30 Animals divided into 5 groups (6 rats/each):
1. Control naïve group; rats were injected with 0.9% NaCl
2. HPCL group; rats were injected intraperitoneally (IP) with DEN & 2AAF 100 mg/kg once weekly for 3 consecutive weeks followed by 1 week rest period then 2AAF was injected once intraperitoneally in a dose of 300mg/kg
3. Pantoprazole 100mg group; HPCL group treated with 100mg/kg pantoprazole i.p.
4. Pantoprazole 50mg group; HPCL group treated with 50mg/kg pantoprazole i.p
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5. Pantoprazole 25mg group; HPCL group treated with 25mg/kg pantoprazole i.p
Pantoprazole was injected 3 times weekly in the last 4 weeks of the experiment group (week 7-10)
At the end of the 10th week rats were anesthetized. Retro orbital blood samples were taken, then rats were sacrificed and liver sample were collected. For each animal, parts of the liver and blood samples were kept in -80 ºC until analysis.
Evaluation of exosomal RAB11A-mRNA, LncRNA-RP11513I15.6 in human and rats’ samples was performed by real time PCR in relation to ACTB as the housekeeping gene in all samples. And exosomal miRNA1262 was performed also by real time PCR in relation to RNU-6 as an internal control in all samples. Then the results were statistically analyzed by the SPSS software.
A significant difference was observed in the positivity rates for exosomal lncRNA-RP11-513I15.6, exosomal miR-1262, and exosomal RAB11A mRNA, which were 96.7%, 95%, and 75%, respectively, in the malignant group. While among the CHC patients, the positivity rates of exosomal lncRNA-RP11-513I15.6, exosomal miR-1262, and exosomal RAB11A mRNA were 7.1%, 28.6% and 31%, respectively