Overexpression of SPARC has been documented in several types of solid tumors, such as breast[7], prostate[8], melanoma[9] and glioblastomas[10]. In contrast, lower levels of SPARC expression have been found in other types of cancers, such as ovarian[11], colorectal[12], pancreatic[13, 14] and acute myelogenous leukemia[15]. These observations suggest that tumorigenic effect of SPARC is cell type specific and may be dependent of the selleck products tumor cell surrounding environment. The

knowledge about SPARC functions in gastric cancer cells is still sparse. Overexpression of the SPARC gene was observed in human gastric cancer in five other reports[16–20]. However, all above-mentioned studies had no detail in gastric cancer cell lines and carcinogenic mechanism. SPARC selleck chemical has been associated with aggressive stages of gastric cancer and is correlated with poor prognosis[16], which suggests that the reduction of SPARC expression may have therapeutic benefit. Indeed, expression of antisense

oligonucleotides against SPARC in melanoma cells blocked tumor formation[21]. The precise biological and molecular mechanisms through which a reduction in SPARC expression might contribute to improved tumor therapy remain to be investigated. Therefore, the aim of the present study was to characterize SPARC functions in gastric cancer cells and explore its possibly carcinogenic mechanism. Materials and methods Cell culture Human Fossariinae gastric cancer cell lines NCI-N87, SGC7901, MGC803, BGC823, HGC27 were Selleck 17-AAG obtained from the Cancer Institute of Chinese Academy of Medical Science. All cells were grown in RMPI 1640 (GIBCO™)medium supplemented with 10% fetal bovine serum, penicillin G (100 units/ml), and streptomycin (100 μg/ml) termed complete medium. Cells were maintained in monolayer culture at 37°C in humidified air with 5% CO2. Chemicals and reagents EDTA-2 sodium, acridine orange, ethidium bromide (EB) and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazoliumbromide (MTT) were purchased from Sigma (St Louis, MO, USA). Mouse monoclonal antibody specific to β-actin was from Sigma. Rabbit polyclonal antibodies specific to Bcl-2 (sc-492), caspase-3 (sc-7148) and PARP (sc-7150) were

bought from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse monoclonal antibodies specific to SPARC(sc-74295) and Bax (sc-7480) were obtained from Santa Cruz Biotechnology. Goat anti-rabbit (w3960) and anti-mouse (w3950) secondary antibodies were purchased from Promega (Madison, WI, USA). RNAi and transfection Human SPARC siRNA and control siRNA were from Dharmacon Bioscience Corp (Chicago, IL, USA). Equimolar amounts of siRNAs were used as per the manufacturer’s instructions with control non-targeting siRNA (CTRL). 150 000 cells were plated per six-well in DMEM with 10% FBS and were allowed to attach overnight. Equimolar amounts of siRNAs were incubated with TransIT-TKO Transfection Reagent from Mirus (Madison, WI, USA) as per the manufacturer’s instructions.