Abstract
Purpose: Human hepatocellular carcinoma is one of the most common causes of death in the world. Metformin and rapamycin may decrease the expression of VEGF protein and subsequently angiogenesis. The purpose of this study was to evaluate the effect of these two drugs on expression of VEGF protein and the cell proliferation in the hepatocellular carcinoma cell line (ATCC HB-8065).Methods: HepG2 was cultured in RPMI-1640 medium at 37°C for 48h as a pre-culture and then treated by different concentrations of metformin (0, 5, 10 and 20 mM) and rapamycin (0, 5, 10 and 20 nM) at different times (12, 24 and 48 h). Cell viability was assessed by the MTT assay. Total RNA was extracted by the Trizol reagent and VEGF gene expression was analyzed by quantitative real-time PCR and was calculated by 2–ΔCt method. The VEGF protein level was determined by Elisa assay. Finally, Apoptosis index was calculated by DAPI staining.Results: Metformin and rapamycin significantly decrease cancer cells viability (p<0.05). Rapamycin but not metformin decreases VEGF gene expression in HepG2 cells. Metformin and rapamycin significantly induce cell apoptosis in hepatocellular carcinoma (HCC) cells.Conclusion: Metformin and rapamycin have an anti-tumor effect on HCC. According to our data rapamycin might have an anti-angiogenesis effect via inhibition of VEGF expression. Our results provide an insight into future clinical strategies to improve chemotherapy outcomes in HCC.
Highlights
Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver and is the fourth leading cause of cancer-related death in the world.[1]
Metformin and rapamycin decrease cancer cells viability To evaluate the effects of metformin and rapamycin on HCC cells growth in vitro, cell viability of HepG2 Cells was assessed by MTT assay following treatment with metformin and rapamycin in various concentrations and at different times
Results showed that both metformin and rapamycin inhibited the growth of HepG2 cells in a dose-dependent manner (p < 0.05) (Figure 1)
Summary
Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver and is the fourth leading cause of cancer-related death in the world.[1]. Angiogenesis is a complex process that is based on cooperation between various cells, such as pericytes, endothelial cells, fibroblasts and smooth muscle cells These cells produce a variety of cytokines and growth factors that interact with other cells or with the extracellular matrix and affect migration, proliferation and angiogenesis.[5] Angiogenesis plays a key role in tumor growth, progression and metastasis.
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