الفهرس 
Title : Molecular assessment of anti-cancer activity of crude Bee Venom and its active component, Melittin, either alone or in combination with Sorafenib in treatment of Hepatocellular carcinoma. In-vitro and in-silico study
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Abstract
Background: Hepatocellular carcinoma (HCC) is the sixth most common tumor worldwide and the second predominant cause of cancer-related death. The standard treatment for advanced HCC is Sorafenib (Sorf), which is a multikinase inhibitor and mediates its action through suppressing angiogenesis along with induction of oxidative stress. However, it has very harsh side effects and can trigger drug-resistance. Thus, there are current attempts to find a safe substitute or adjuvant for Sorf in treating HCC. Recently, Bee Venom (BV) and its active component, Melittin (Mel), have demonstrated several therapeutic effects on various types of malignant cells through different mechanisms such as induction of apoptosis, suppression of proliferation, migration and cell cycle progression. Aim of Study: To determine the affinity of Mel towards different cellular markers in-silico; to evaluate the anti-proliferative activity of BV or Mel either as a single treatment or when combined with Sorf against HCC cell line, HepG2; to determine the safety of these compounds on the normal liver cell line, THLE-2; and to investigate the mechanism of action of single and combined treatments in terms of gene expression, oxidative stress and cell cycle analysis.Methods: Interactions between Mel and some cancer-related proteins were evaluated computationally using molecular docking tools. The cytotoxicity of single treatments of BV, Mel and Sorf on HepG2 and THLE-2 cells was conducted using MTT assay to determine the IC50. Combinations of Sorf with BV or Mel was performed in 24 non-constant ratios on HepG2 cells and combination Index (CI) was calculated to determine the interaction type. Gene expression of certain markers that are involved in apoptosis, cell cycle, migration, metastasis, hypoxia, angiogenesis and inflammation were measured using real-time PCR. Status of oxidative stress and antioxidant enzymes were evaluated spectrophotometrically. Cell cycle analysis using flow cytometry was conducted.Results: Computational analysis showed that Mel can bind to and inhibit some critical proteins in cancer development. Single treatments of BV, Mel and Sorf showed IC50 of 2.8, 1.4 and 4.15 og/ml on HepG2 versus 95.64, 78.31 and 4.45 og/ml, respectively on THLE-2. Among 24 combinations, the most effective ratios were BV: Sorf (3/2: 3/4) and Mel: Sorf (3: 3/4) on HepG2. A significant increase in the expression level of apoptotic markers and a significant decrease in oncogenic markers were observed, while the oxidative stress markers showed insignificant changes. The treatments caused cell cycle arrest at G2/M phase.