الفهرس 
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Abstract
Summary
HCC is one of the most common cancer types
worldwide, with a high prevalence and a very low survival rate
of 3 – 5 %. Many factors play a role in the etiology of HCC,
but, most importantly, HCC can be caused by HBV and HCV infections. HCC cells, unlike normal cells, are resistant to deathreceptor-mediated apoptosis because their surface death
receptors are cross-linked with either agonistic antibodies or
soluble death ligand proteins. Several cell cycle proteins play a
role in the etiology of liver cancer.
So far, the development of chemotherapeutic drugs has
targeted molecules in HCC, but the results have not been
promising because of drug resistance and systemic toxicity,
specifically in the advanced stage of this deadly malignancy.
Therefore, searching for new natural and nontoxic compounds
with cytotoxic effects against HCC cells is of particular
interest.One of the less studied flavonoids is hesperidin, the most
abundant flavanone in citrus fruits. Hesperidin possesses
medical benefits, including antioxidant, anti-inflammatory,
hypotensive properties and anti-carcinogenic activities This study was carried out in order to evaluate the
antiproliferative effect of hesperidin on human hepatocellular
carcinoma HepG2 cell line which is a well-differentiated
transformed cell line closely related to HCC. In this study, The HepG2 cells was maintained in RPMI-
1640 medium was supplemented with 10% heat inactivated
FBS, 2% Penecillin and streptomycin, 1 % Amphotricin B,
culture medium was changed three times per week. Hesperidin was dissolved in DMSO at room temperature
to prepare 1mM stock solution of hesperidin; this solution was
diluted with tissue culture medium before treatment of the
cells. Then, completely confluent HepG2 cells in T-25 flasks
were treated with different concentrations of hesperidin 25, 50
& 100 μM and untreated cells serve as control. All flasks were
maintained at 37°C in a humidified, incubator with 95% air and
5 %CO2.After 24, 48 & 72 hrs all flasks were trypsinized with 2
ml of trypsin-EDTA and counted under the light microscope
(100x) using trypan-blue dye (0.04%) to count the number of
viable cells. The cell suspensions were centrifuged then, the
pellet were washed with PBS and kept in sterile Eppindorffs at
-80 °C until the biochemical assays were performed.
RT-PCR was performed to detect NF-kB geneexpression, as the following: total RNA was extracted from all
of the above cell-lysate, and then, in order to preserve RNA
samples, total RNA was transcribed into cDNA. RT-PCR was
performed using a real time PCR 7500 fast thermal cycler.
At 24, 48 & 72 hrs time intervals after treatment, culture
medium was harvested for measuring the content of AFP using
Abbott AxSYM Immunology Analyzer based on MEIA
technology, untreated cells serve as control.
Results obtained could be summarized as follows:
1- Hesperidin inhibited the growth and caused
ultrastructural morphological changes in HepG2 cells in
a time-dependent manner at 24, 48 & 72 hrs after
treatment as the % of survival rates have significantly
decreased with the progress of the time compared with
the untreated control cells.
2- Hesperidin inhibited the activity of NF-kB gene
expression of HepG2 treated-cells compared with the
control.
3- Hesperidin inhibited AFP secretion in HepG2 cells
compared with the control.
Therefore, the conclusion was that hesperidin inhibits the
growth of HepG2 cells through suppressing the activity of NFkB
gene expression and modulates the secretion of AFP from
the cells.