ABSTRACT
Hepatocellular carcinoma (HCC) is the most common primary liver cancer and is one of the leading causes of cancer-related deaths worldwide. Imatinib and GNF-5 are breakpoint cluster region-Abelson murine leukemia tyrosine kinase inhibitors which have been approved for the treatment of chronic myeloid leukemia and various solid tumors. However, the effect and underlying mechanisms of imatinib and GNF-5 in HCC remain poorly defined. In this study, we investigated the anticancer activity and underlying mechanisms of imatinib and GNF-5 in HepG2 human hepatocarcinoma cells. Cell proliferation and anchorage-independent colony formation assays were done to evaluate the effects of imatinib and GNF-5 on the growth of HepG2 cells. The cell cycle was assessed by flow cytometry and verified by immunoblot analysis. Gene overexpression and knockdown assays were conducted to evaluate the function of S-phase kinase-associated protein 2 (Skp2). Imatinib and GNF-5 significantly inhibited the growth of HepG2 cells. Imatinib and GNF-5 induced G0/G1 phase cell cycle arrest by downregulating Skp2 and upregulating p27 and p21. Overexpression of Skp2 reduced the effect of imatinib and GNF-5 on HepG2 cells. Knockdown of Skp2 suppressed the proliferation and induced G0/ G1 phase arrest. Furthermore, knockdown of Skp2 enhanced the effect of imatinib and GNF-5 on growth of HepG2 cells. In conclusion, imatinib and GNF-5 effectively suppress HepG2 cell growth by inhibiting Skp2 expression. Skp2 promotes the cell proliferation and reverse G0/G1 phase cell cycle arrest and it represents a potential therapeutic target for HCC treatment.
ABSTRACT
Hepatocellular carcinoma (HCC) is the most common primary liver cancer and is one of the leading causes of cancer-related deaths worldwide. Imatinib and GNF-5 are breakpoint cluster region-Abelson murine leukemia tyrosine kinase inhibitors which have been approved for the treatment of chronic myeloid leukemia and various solid tumors. However, the effect and underlying mechanisms of imatinib and GNF-5 in HCC remain poorly defined. In this study, we investigated the anticancer activity and underlying mechanisms of imatinib and GNF-5 in HepG2 human hepatocarcinoma cells. Cell proliferation and anchorage-independent colony formation assays were done to evaluate the effects of imatinib and GNF-5 on the growth of HepG2 cells. The cell cycle was assessed by flow cytometry and verified by immunoblot analysis. Gene overexpression and knockdown assays were conducted to evaluate the function of S-phase kinase-associated protein 2 (Skp2). Imatinib and GNF-5 significantly inhibited the growth of HepG2 cells. Imatinib and GNF-5 induced G0/G1 phase cell cycle arrest by downregulating Skp2 and upregulating p27 and p21. Overexpression of Skp2 reduced the effect of imatinib and GNF-5 on HepG2 cells. Knockdown of Skp2 suppressed the proliferation and induced G0/ G1 phase arrest. Furthermore, knockdown of Skp2 enhanced the effect of imatinib and GNF-5 on growth of HepG2 cells. In conclusion, imatinib and GNF-5 effectively suppress HepG2 cell growth by inhibiting Skp2 expression. Skp2 promotes the cell proliferation and reverse G0/G1 phase cell cycle arrest and it represents a potential therapeutic target for HCC treatment.
ABSTRACT
Obesity is a risk factor for various diseases, including cardiovascular disease, diabetes, renal disease, hypertension, cancer, and neural disease. Adipose tissue in animals is important for the mobilization of lipids, milk production, deposition of fat in different depots, and muscle and meat production. Understanding the genetic and physiological causes of metabolic disease is a priority in biomedical genome research. In this study, we examined several variables in mice fed a high-fat diet, including serum composition, body weight, total calorie intake, and differentially expressed genes. Body weight and blood glucose levels were not significantly different between animals fed high-fat and normal diets. However, high-fat diet groups showed reduced calorie and food intakes. Levels of sodium, ionized calcium, glucose, hematocrit, hemoglobin, pH, PCO2, PO2, TCO2 +, HCO3 +, base excess, and SO2 in the blood were not significantly different between mice fed high-fat and normal diets. Serum potassium concentration, however, was lower in mice a high-fat diet. Differentially expressed genes were also compared between the two groups. The purpose of this study was to discover new genes as a result of annealing control primer (ACP) PCR using 20 random primers. Five down regulated genes were identified and three of others were upregulated by high-fat diet. Known genes were excluded from this result. In addition, the relationships among candidate genes and high-fat diet should be investigated according to potassium concentration in the blood. In conclusion, mice fed normal and high-fat diets showed no significant difference in body weight, whereas high-fat diet led to changes in blood composition and differential expression of several genes. These findings may provide a better understanding of the mechanisms underlying the association between obesity and metabolic diseases.