Expression analysis of URG4 (upregulated gene 4) in osteoblast-like cells, Saos-2 and MG63

Abstract

URG4 is a novel gene firstly found in uninfected liver cells stimu-lated by HBxAg (Hepatitis B virus encoded X antigen) and HCC(Hepatocellular Carcinoma) cells. It is associated with cellgrowth, growth factor-independent survival, and anchorage-inde-pendent growth in HepG2 cells. It has been implicated as a puta-tive oncogene contributing HBxAg related multi-stephepatocarcinogenesis.URG4 has also been shown to be strongly expressed in gastriccancers and osteosarcoma tissues that is the most common pri-mary malignant bone tumor by immunohistochemistry. There-fore, in the present study, we aimed to determine the expressionprofile of URG4 in two different osteoblast like cell lines; MG63and Saos-2. These two osteoblastic cells represent different cellcharacteristics. Saos-2 cells are similar with late osteoblastic dif-ferentiation stage in human cells and exhibit the entire differenti-ation sequence of osteoblastic cells that form a calcifying matrixin diffusion chambers. MG63 cell line presents osteoblast like.URG4 expression level was determined by RT-PCR strategy.Cells were cultured in DMEM (Dulbecco’s Modified Eagle Med-ium) containing 10% FCS in humidified incubator. Total RNAwas isolated from cultured cells was used to synthesize cDNA.URG4 was amplified by using specific primers. It was visualizedby agarose gel electrophoresis and photographed. URG4 mRNAwas present in both cell lines. Further studies will be carried outif there is any effect of some cytokines on these cell lines onURG4 gene expression.