Amr Amin

Publications (48) View all

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  Article: The anticancer effect of saffron in two p53 isogenic colorectal cancer cell lines.

  Khuloud Bajbouj, Jan Schulze-Luehrmann, Stefanie Diermeier, Amr Amin, Regine Schneider-Stock
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  ABSTRACT: BACKGROUND: Saffron extract, a natural product, has been shown to induce apoptosis in several tumor cell lines. Nevertheless, the p53-dependency of saffron's mechanism of action in colon cancer remains unexplored. METHODS: In order to examine saffron's anti-proliferative and pro-apoptotic effects in colorectal cancer cells, we treated two p53 isogenic HCT116 cell lines (HCT wildtype and HCT p53-/-) with different doses of the drug and analyzed cell proliferation and apoptosis in a time-dependent manner. MTT viability and crystal violet assays were performed in order to determine the effective dose of saffron on both cell lines. The cell cycle progress was examined by Flow cytometric analysis. Apoptosis was assessed using Annexin-PI-staining and Western Blotting for caspase 3 and PARP cleavage. Autophagy was determined by Western Blotting of the light chain 3 (LC3)-II and Beclin 1 proteins. The protein content of phospho-H2AX (gammaH2AX), a sensor of DNA double strand breaks, was also analyzed by Western Blotting. RESULTS: Saffron extract induced a p53-dependent pattern of cell cycle distribution with a full G2/M stop in HCT116 p53 wildtype cells. However, it induced a remarkable delay in S/G2 phase transit with entry into mitosis in HCT116 p53 -/- cells. The apoptotic Pre-G1 cell fraction as well as Annexin V staining and caspase 3 cleavage showed a more pronounced apoptosis induction in HCT116 p53 wildtype cells. Obviously, the significantly higher DNA-damage, reflected by H2AX protein levels in cells lacking p53, was coped by up-regulation of autophagy. The saffron-induced LC3-II protein level was a remarkable indication of the accumulation of autophagosomes, a response to the cellular stress condition of drug treatment. Conclusions: This is the first study showing the effect of saffron in HCT116 colorectal cancer cells with different p53 status. Saffron induced DNA-damage and apoptosis in both cell lines. However, autophagy has delayed the induction of apoptosis in HCT116 p53 -/- cells. Considering the fact that most tumors show a functional p53 inactivation, further research is needed to elucidate the long-term effects of saffron in p53 -/- tumors.
  BMC Complementary and Alternative Medicine 05/2012; 12(1):69. · 2.24 Impact Factor
• Article: Neural Network Assessment of Herbal Protection against Chemotherapeutic-Induced Reproductive Toxicity

  Amr Amin, Doaa Mahmoud-Ghoneim, Muhammed I Syam, Sayel Daoud
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  ABSTRACT: The aim of this study is to assess the protective effects of Ginkgo biloba’s (GB) extract against chemotherapeutic-induced reproductive toxicity using a data mining tool, namely Neural Network Clustering (NNC) on two types of data: biochemical & fertility indicators and Texture Analysis (TA) parameters. GB extract (1 g/kg/day) was given orally to male albino rats for 26 days. This period began 21 days before a single cisplatin (CIS) intraperitoneal injection (10 mg/kg body weight). GB given orally significantly restored reproductive function. Tested extract also notably reduced the CIS-induced reproductive toxicity, as evidenced by restoring normal morphology of testes. In GB, the attenuation of CIS-induced damage was associated with less apoptotic cell death both in the testicular tissue and in the sperms. CIS-induced alterations of testicular lipid peroxidation were markedly improved by the examined plant extract. NNC has been used for classifying animal groups based on the quantified biochemical & fertility indicators and microscopic image texture parameters extracted by TA. NNC showed the separation of two clusters and the distribution of groups among them in a way that signifies the dose-dependent protective effect of GB. The present study introduces the neural network as a powerful tool to assess both biochemical and histopathological data. We also show here that herbal protection against CIS-induced reproductive toxicity utilizing classic methodologies is validated using neural network analysis.
  Theoretical Biology and Medical Modelling. 01/2012; 9(1).
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  Article: Cancer chemoprevention.

  Amr Amin, Metka Filipič, Su S Chen, Regine Schneider-Stock
  Journal of Biomedicine and Biotechnology 01/2012; 2012:250491. · 2.44 Impact Factor
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  Article: Saffron: a potential candidate for a novel anticancer drug against hepatocellular carcinoma.

  Amr Amin, Alaaeldin A Hamza, Khuloud Bajbouj, S Salman Ashraf, Sayel Daoud
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  ABSTRACT: Saffron has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of saffron against diethylnitrosamine (DEN)-induced liver cancer in rats. Administration of saffron at doses of 75, 150, and 300 mg/kg/day was started 2 weeks prior to the DEN injection and was continued for 22 weeks. Saffron significantly reduced the DEN-induced increase in the number and the incidence of hepatic dyschromatic nodules. Saffron also decreased the number and the area of placental glutathione S-transferase-positive foci in livers of DEN-treated rats. Furthermore, saffron counteracted DEN-induced oxidative stress in rats as assessed by restoration of superoxide dismutase, catalase, and glutathione-S-transferase levels and diminishing of myeloperoxidase activity, malondialdehyde and protein carbonyl formation in liver. The results of immunohistochemical staining of rat liver showed that saffron inhibited the DEN-mediated elevations in numbers of cells positive for Ki-67, cyclooxygenase 2, inducible nitric oxide synthase, nuclear factor-kappa B p-65, and phosphorylated tumor necrosis factor receptor. Saffron also blocked the depletion in the number of cells positive for TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling) and M30 CytoDeath in liver tissues of DEN-treated rats. In vitro experiments carried out using HepG2 cells also confirmed these findings and showed inhibition of nuclear factor-kappa B activation, increased cleavage of caspase-3, as well as DNA damage and cell cycle arrest upon saffron treatment. Conclusion: This study provides evidence that saffron exerts a significant chemopreventive effect against liver cancer through inhibition of cell proliferation and induction of apoptosis. This report also shows some evidence that saffron protects rat liver from cancer via modulating oxidative damage and suppressing inflammatory response.
  Hepatology 05/2011; 54(3):857-67. · 11.66 Impact Factor
• Article: Texture analysis of liver fibrosis microscopic images: a study on the effect of biomarkers.

  Amr Amin, Doaa Mahmoud-Ghoneim
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  ABSTRACT: Chronic hepatic injury results in liver fibrosis with eventual progression to irreversible cirrhosis. Liver fibrogenesis involves the activation of the quiescent hepatic stellate cell into an activated myofibroblast that is characterized by α-smooth muscle actin (α-SMA) expression and the production of collagens (types I and III). In the present study, rats were randomly divided into three groups: (i) control group, where rats were only treated with a vehicle; (ii) fibrosis group, where rats were treated with carbon tetrachloride (CCl(4)) to induce liver fibrosis; and (iii) silymarin group, where rats were protected with silymarin during CCl(4) treatment. Rats were sacrificed and sections of liver tissue were counterstained with hematoxylin and eosin and Masson's trichrome. Other sections were immunostained using collagens and α-SMA primary antibodies. Fibrosis was confirmed using serum marker measurements. Microscopic images of the stained sections were acquired and digitized. The Biomarker Index of Fibrosis (BIF) was calculated from the images by quantifying the percentage of stained fibers. Statistical methods of texture analysis (TA), namely co-occurrence and run-length matrices, were applied on the digital images followed by classification using agglomerative hierarchical clustering and linear discriminant analysis with cross validation. TA applied on different biomarkers was successful in discriminating between the groups, showing 100% sensitivity and specificity for classification between the control and fibrosis groups using any biomarker. Some classification attempts showed dependence on the biomarker used, especially for classification between the silymarin and fibrosis groups, which showed optimal results using Masson's trichrome. TA results were consistent with both BIF and serum marker measurements.
  Acta Biochimica et Biophysica Sinica 03/2011; 43(3):193-203. · 1.38 Impact Factor