Background: Hyperglycemia is one of the important featurs of diabetes. In cell culture studies different methods are used to mimic the hyperglycemia condition. In this study we investigate response of human liver cancer cell line (HEPG2) to high insulin, high glucose, and high insulin/ high glucose medium exposure. Methods: HEPG2 cells were settled in DMEM+0. 1% FBS or DMEM free-serum medium with high concentrations of d-glucose (30 mm) and/or insulin (1μ M) for 24h after an overnight starving in serum-free medium. The level of hyperglycemia was estimated in the supernatants via GOD-POD method. Results: Serum-free madium with high insulin/ high glucose consentration made the higher level of hypreglycemia in HEPG2 cells. Conclusions: Our study interduced high insulin/ high glucose treatment as the best way to induction hyperglycemia.

Aim: Addiction is an important social and health problem in many Middle Eastern countries. Studies show that opium is the most commonly used substance after tobacco in many countries, especially in Iran. Opium is obtained from the seeds of the Papaver somniferum plant and contains more than 40 different alkaloids, the most important of which are morphine, codeine, papaverine, noscapine, and thebaine. Various studies show that morphine, which is one of the most important substances in opium, increases the production of free radicals in the body and reduces the antioxidant capacity. Opium consumption has various adverse health effects on the body. Therefore, long-term use of this combination can be related to some pathological consequences, including neurological disorders, liver toxicity, kidney dysfunction, oxidative stress, and apoptosis. Most drugs are metabolized by liver hepatocytes and excreted by kidney cells. Therefore, opium consumption can directly cause damage to liver cells. Free radicals play an important role in liver diseases. The increase of free radicals and on the other hand the decrease of antioxidants in the body caused liver toxicity. Oxidative stress is caused by an imbalance between the production of free radicals and their neutralization inside the body by antioxidant defense mechanisms. Oxidative stress is produced in various diseases, consumption of toxic substances, and old age, since it is not possible to investigate the effect of opium on human liver cells, and no study has been conducted in this field. The liver is the most important organ that is directly responsible for the metabolism and excretion of opium. Therefore, in this study, the effects of opium on oxidative stress markers in HEPG2 cell line were determined.Material and methods: Human liver cancer cell lines (HEPG2) were used in this experimental study. At first, the cell line was placed in a 25 ml flask in the culture medium. DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS (Fetal bovine serum), and 1% penicillin and streptomycin antibiotics were incubated in a 37°C incubator with 5% CO2 . The cells were treated with different concentrations of opium (0-100ug/ml) for 24 hours. Then IC50 was determined and then a lower concentration, IC50, and a higher concentration (three concentrations in total) were used for further studies.The lipid peroxidation was measured by thiobarbituric acid method. The total oxidant was measured using xylenol orange. The amount of total antioxidants was determined by the FRAP (Ferric Reducing / Antioxidant Power) method. The catalase, glutathione peroxidase, and superoxide dismutase enzyme activities were measured by colorimetry methods. the data was entered into SPSS software (Version 20)  and analyzed using ANOVA and Tukey statistical tests. P less than 0.05 was considered a significant level. Results: Statistical analysis results indicated a significant difference in the amount of oxidative stress factors compared with control (p<0.001). In the group receiving opium with a dose of 60 and 70 mg/ml compared to the control group, the amount of TOS and MDA increased significantly (p<0.001), while the amount of TAC decreased (p<0.001). Also, the activity of catalase, glutathione peroxidase, and superoxide dismutase enzymes decreased in opium-treated groups (p<0.001). Conclusion: The results of this research showed that opium had a lethal effect on HEPG2 cells. Also, opium caused an increase in oxidative stress in this cell line, which indicates the vulnerability of the liver against this compound. The results of this study show that although opium has analgesic effects, it can seriously damage the liver tissue.

Background: Factor VII, is a coagulant protease; it begins the proteolytic cascade reactions and produces thrombin. The use of recombinant human factor VII, (rhFVII) is effective for the treatment of patients with hemophilia A or B. It is a target for gene therapy. This study was done to clone factor VII from HEPG2 cell line.Methods: RNA was extracted from the hepatoma, (HEPG2), cell line. On reverse transcription FVII cDNA was amplified by RT-PCR. PCR product was cloned into the pTZ57R/T vector and transported into the E-coli cells.Results: By amplification of the FVII gene, the PCR band was observed and cloning into the vector was confirmed by restriction analysis.Conclusion: In this paper we report the cloning of factor VII from HEPG2 cell line.

Objective: The objective of the present study was to evaluate the cytotoxic potentials of Evolvulus alsinoides in human hepatoma HEPG2 cells. Materials and Methods: HEPG2 cells were treated with methanolic extract of E. alsinoides at 20, 40 and 80 μ g/ml for 24 hr and cytotoxic effect was analyzed by MTT assay. The apoptosis rate was investigated by Hoechst 33342 and annexin V/propidium iodide staining. Mitochondrial membrane potential was evaluated by rhodamine staining. Also, the expression of catenin – β 1 protein was analyzed by western blotting. Results: E. alsinoides methanolic extract treatment caused significant cytotoxicity in HEPG2 cells in a concentration-dependent manner. Dual staining assay confirmed the presence of early and late apoptotic cells only in extract-treated groups. Plant extract treatment also caused nuclear fragmentation and chromatin condensation in HEPG2 cells. Mitochondrial membrane potential also reduced upon E. alsinoides treatments. This treatment also modulated the catenin – β 1 protein expression. Conclusion: In this study, we demonstrated the proapoptotic potential E. alsinoides in HEPG2 cells; thus, this plant may be beneficial in the treatment of liver cancer.