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Vol.9 No.4-4: MiRNA-122 association with TNF-α in some liver diseases of Egyptian patients

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By: Ahmed Abdelhalim Yameny1, Sabah Farouk Alabd1, and Magda Ahmed M. Mansor2

1Molecular Biology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), University of      Sadat City, Egypt

2Department of Histology, Faculty of Medicine, Menoufia University, Egypt

Abstract:

Background:  Due to the high frequency of HCC, ongoing research is needed to find precise, non-invasive biomarkers for early identification and follow-up that will improve prognostic results. Patients and methods: this study was conducted on 90 patients with liver diseases and 25 healthy control G1, patients divided into 4 groups, (G2) 25 patients with HCV infection, (G3) 25 HCC+HCV infection, (G4) 25 patients with HBV infection, (G5) 15 patients with HCC + HBV. Results: Serum miR-122 and TNF-α levels were increased in HCV and HBV infection significantly with p-value >0.001*compared to the control group, and their levels decreased when developed into HCC but still higher than the healthy subjects significantly with p-value >0.001. For discriminating HCV from HCV+HCC the cut-off for miR-122 was >7.1 at sensitivity 100%, specificity 100%, and the AUC was 1.0 (Excellent) P-value <0.001, also the sensitivity and specificity for TNF-α 72%, and 60% respectively with cut off >12.1 and AUC of 0.745 (Good) p-value 0.003. For discriminating HBV from HBV+HCC the cut-off for miR-122 was ≤6.4 at a sensitivity of 86.67% and specificity of 96%, and the AUC of miR-122 was 0.99 (Excellent) P-value <0.001, also the sensitivity and specificity for TNF-α 93.33%, and 48.0% respectively with cut-off ≤15.73, TNF-α has AUC of 0.527 (fair) it was not significant p-value 0.780.

MiRNA-122-association-with-TNF-α-in-some-liver-diseases-of-Egyptian-patients

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Vol.9 No.3-2:Biomolecular evaluation of apoptosis, cell cycle, oxidative stress, and limiting enzymes of the glycolytic pathway in hepatocellular carcinoma cell line HepG2 treated with crude snake venom with or without sorafenib

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By: Maha Y. Abdel Al Shakour1, Emad M. Elzayat2*, Khalid M. Mahmoud3,

Mamdouh I. Nassar4, Abdel Hamid Z. Abdel-Hamid5

1PhD student at Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

2* Biotechnology Department, Faculty of Science, Cairo University, Giza, Egypt

3Pharmacognosy Department, NRC, Giza, Egypt

4Entomology Department, Faculty of Science, Cairo University, Giza, Egypt

5Therapeutic Chemistry Department, NRC, Giza, Egypt

Abstract

Background: Natural venoms have biological activities including anti-inflammatory, antimicrobial, and anticancer effects.  Hepatocellular carcinoma (HCC) is still a worldwide problem and difficult to treat by chemotherapeutic agents especially sorafenib (SOR), as it evokes many harsh side effects and is disable to differentiate between normal and cancer cells. Objective: The present study aimed to test the hypothesis that combining crude venoms of the snake or the bee or the scorpion could synergistically enhance the antiproliferative effects of SOR in hepatocellular carcinoma cell line (HepG2). Experimental design: Separate crude venoms have been applied to HepG2 cells and normal human retinal cells(RBE1) for estimation of IC50. The most effective venom has been combined with sorafenib in five nonconstant ratios and the combination index (CI) was estimated to expose their synergistic or antagonistic action. The best combination was used for downstream analysis. Results: The crude snake venom exhibited the most cytotoxic effect and the least IC50. It has been combined with sorafenib, and the combination index (CI) was calculated. IC25 SV + IC10 SOR was the best combination with CI=0.209 indicating high synergistic cytotoxic activity against HepG2. The underlining molecular mechanisms of action, in terms of the expression level of apoptotic genes (p53, Bax, Caspase 3, and Bcl2), flow cytometric analysis of cell cycle, oxidative stress markers as well as the activity of some limiting enzymes in the glycolytic pathway (ALDOB, PK and LDH) have been investigated.

Conclusion: Our results suggest a novel synergistic, and anti-proliferative effect of snake venom with sorafenib on HepG2 cells.

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Vol.7 No.4 – 4: Therapeutic Role of Quercetin against Experimentally Induced Hepatocellular Carcinoma in Female Albino Rats and their offspring

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By: Asma R Abdeltawab1, Hassan I.H Elsyyad2, Karoline K.Abdelaziz1,

Abd El-Fattah B. M. El-Beltagy1*

1, Zoology Department, Faculty of Science, Damanhour University, Damanhour, Egypt

2Zoology Department, Faculty of Science, Mansoura University, Mansoura, Egypt

Abstract

Aim:  To evaluate the potential role of quercetin against N-methyl nitrosourea (MNU)-induced hepatocellular carcinoma (HCC) in pregnant rats and their offspring. Material & Methods: Twenty-four female rats were used in this study. Six rats were preserved without treatment and the other eighteen female rats were induced by a single dose of MNU (50 mg /kg B wt). After confirmation of the positive tumor marker test, female rats were placed with the males for mating. The pregnant rats were divided into four groups (n=6). Group1: control rats, group2: Quercetin supplemented rats (20 mg/kg B.Wt, group3: MNU-induced rats, and group4: MNU-treated rats followed by supplementation with quercetin. At the end of the weaning period, the mothers and their offspring (at 21 days old postnatal) of all groups were sacrificed, the liver was removed immediately for histological and immunohistochemical investigations. Also, blood samples were collected, centrifuged, and processed for the estimation of antioxidants. Results: In the control and quercetin groups, the histological investigation of the liver of mother rats and their offspring appeared with normal architecture. In Group3 (MNU-induced group) the liver sections of mother’s rats revealed degenerated hepatic lobules with pronounced cellular hyperplasia (HCC) especially around the central vein and portal area as well as numerous Kupffer cells and fat droplets.  However, the liver sections of offspring displayed little cellular hyperplasia but the central and portal veins appeared damaged and congested with blood. Immunohistochemically, the liver sections of MNU-induced mother’s rats and their offspring appeared strongly stained with α-FP antibody and negatively stained with caspase-3 antibody. Furthermore, the levels of serum antioxidants; SOD, CAT&GSH were significantly decreased however the levels of MDA and NO were significantly increased in MNU-induced mother rats and their offspring if compared with control. In group 4, quercetin was able to restore the histological and immunohistochemical changes in the liver induced by MNU.  Also, the levels of antioxidants, as well as MDA and NO, were markedly restored near to the normal level as in control.  Conclusion: Quercetin has a powerful therapeutic role against MNU-induced HCC in pregnant rats and their offspring.

Therapeutic

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Vol.5 No.3 – 5 : Serum cytokine profile during disease progression stages in male and female hepatitis C patients

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By: Abdel-Rahman N. Zekri1, Abeer M. Badr2*, Shimaa Rabah2, Maysa El Razky3, Somaya El Deeb2

1Molecular Virology and Immunology Unit, Department of Cancer Biology, National Cancer Institute, Cairo University, Cairo 11976, Egypt.

2Zoology Department, Faculty of Science, Cairo University, Giza, 12613, Egypt.

3Hepatology Department and Tropical Medicine, Faculty of Medicine, Cairo University, Cairo, 11441, Egypt.

Abstract

T helper (Th) cytokines play a key role in the immunological aspects of hepatitis C virus (HCV) pathogenesis. The pattern of Th1 (IL-2, interferon (IFN)-γ), Th2 (IL-10), and immunomodulatory cytokines (IL-12, IL-1β, IFN-α and tumor necrosis factor-α receptor (TNF-αR2) balance participated in the outcome of host immune responses. The study aimed toinvestigate the serum levels of Th1/Th2 and immunomodulatory cytokines in HCV infected patients in both genders during various liver disease stages compared to healthy controls. Blood samples were collected from 16 healthy individuals and 77 patients at different disease stages including chronic, cirrhosis, and hepatocellular carcinoma (HCC). Serum cytokine levels were measured by ELISA. Levels of serum IL-12 and IL-10 were significantly higher in both genders in all groups than those in corresponding healthy subjects. Whereas, HCV infected female patients showed significant lower levels of IL-2, IL-1β, IFN-α in chronic and cirrhosis stages than corresponding males. Serum level of IFN-γ could be utilized as biomarker for early detection of HCC. Finally, cytokine response variation in gender during various stages of disease, imply that the subsequent activation and attenuated functional immune responses displayed differences in the balance of Th1 and immunomodulatory related cytokines between females and males upon infection.

Serum cytokine profile during disease progression stages in male and female hepatitis C patients-converted

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Vol.4 No.4 – 11 : Comparison between glypican-3 and alpha-fetoprotein in discrimination of hepatocellular carcinoma from cirrhotic patients

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By: Abdelfattah Mohamed Attallah*1, Mohamed Abd El-Hafez El-Far 2, Mohamed Mostafa Omran3, Aya Mohamed Saeed1, Mohamed Sayed Elbendary1, Kareem Abdelfattah Attallah1, Khaled Farid Mari4

1* Research & Development Department, Biotechnology Research Center, New Damietta City, Egypt

2 Chemistry department, Faculty of Science, Mansoura University, Mansoura, Egypt

3 Chemistry department, Faculty of Science, Helwan University, Helwan, Egypt

4 Tropical medicine department, Faculty of Medicine, Mansoura University, Mansoura, Egypt

Abstract

Background: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer. Liver cirrhosis progression could be a consequence for developing HCC. Although alpha-fetoprotein (AFP) is widely used as, a marker for detection of HCC, but it has poor sensitivity. Objective: Evaluate the diagnostic power of serum AFP and Glypican-3 (GPC3) as biomarkers of development of HCC. Subjects and Methods: A total of 182 patients, 110 patients with HCC and 72 patients with liver cirrhosis were included. AFP and GPC3 were determined using ELISA. The diagnostic power was evaluated using Area under Roc curve (AUC). Results: levels of AFP and GPC3 in sera of HCC patients were higher than in those with liver cirrhosis (p < 0.0001). AFP had Area under curve (AUC) = 0.772 with sensitivity 39.1%, specificity 97.2%, positive predictive value (PPV) 97.7%, negative predictive value (NPV) 34.3% and efficiency 53.4% while GPC3 had AUC=0.841 yielded sensitivity 76.4%, specificity 86.1%, PPV 94.4%, NPV 64.3% and efficiency 78.8%. There was significant weak correlation (r = 0.241; P < 0.001) between AFP and GPC3. Conclusions: GPC3 is good marker for HCC diagnosis. Therefore, GPC3 may be more useful than AFP in differentiating HCC from cirrhotic patients.

Comparison between glypican-3 and alpha-fetoprotein in discrimination of hepatocellular carcinoma from cirrhotic patients-converted

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Vol.3 No.4 – 2: miRNA-122 from Laboratory biomarker to the treatment of HCV

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By : Ahmed Abdelhalim Yameny

Abstract

     Hepatitis C virus (HCV) is an important human pathogen that infects as many as 185 million persons worldwide, In the long-term, this can lead to advanced liver fibrosis, cirrhosis, and hepatocellular carcinoma HCC, there is currently no vaccine for hepatitis C. About 15–45% of infected persons spontaneously clear the virus within 6 months of infection without any treatment, miRNAs are endogenous short single-stranded noncoding RNAs and they are post-transcriptional negative regulators of gene expression, about18-22 nucleotides long, and play a crucial role in the regulation of gene expression. Now there are over 2500  mature potential human microRNAs recorded in miRBase (version 20, accessed January 2014), 84 miRNAs in serum and plasma of HCV-infected patients to identify miRNAs that correlated with different stages histologically assessed liver disease severity and during HCV infection, miRNA-122 is the most abundant in the liver,miRNA-122 is responsible, for liver homeostasis, several studies showed that miRNA-122 is required for HCV replication in infected cells, it can use as a serum biomarker over alanine leucine transaminase(ALT) in predicting the presence of chronic HCV infection, miR-122 also plays a crucial role in the regulation of cholesterol and fatty acid metabolism in the adult liver, and was identified as a regulator for systematic iron homeostasis, therapies that target it could present an effective approach for the development of new HCV antiviral drugs, Recently, the development of the anti-miR122 therapeutic miravirsen. Miravirsen (formerly SPC3649) is a 15-base oligonucleotide that is complementary to part of miR-122 and is the first miRNA-targeting agent administered to patients. Miravirsen has demonstrated in vitro antiviral activity against all HCV genotypes, Miravirsen interferes with the functions of miR-122 both in viral proliferation and in cholesterol homeostasis, miravirsen has demonstrated broad antiviral activity and a relatively high genetic barrier to resistance.

miRNA-122-from-Laboratory-diagnosis-to-treatment-of-HCV-converted

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