Sgc 7901

The human gastric cancer cell line SGC-7901 was obtained from Shanghai Cell Bank, Chinese Academy of Sciences (Shanghai, China), and was cultured in RPMI 1640 medium (Life Technologies Inc., Grand Island, NY, USA) supplemented with 10% foetal bovine serum plus penicillin (50 IU/ml) and streptomycin (50 μg/ml) in a humidified incubator containing 5 ml/L CO2 at 37°C. The growth medium was refreshed every 2 days.
To manipulate RASSF1A expression in gastric cancer cells, we first cloned RASSF1A cDNA into a pCDA3.1 vector at the BamHI and XhoI sites. After DNA sequence confirmation, the pcDNA3.1-RASSF1A and control pcDNA3.1-vector were stably transfected into the SGC-7901 human gastric carcinoma cell line using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's instructions, and SGC-7901 cells stably expressing the RASSF1A gene were established by G418 selection.
To determine the role of miR-711 in regulating the growth of SGC-7901 cells, we transfected SGC-7901 cells with the miR-711 overexpression vector (miR-711-mimics), miR-711 expression-inhibiting vector (miR-711-inhibitors) or empty vector (NC) (GenePharma Co., Ltd, Shanghai, China) and then assayed the viability of SGC-7901 cells by MTT 24 h, 48 h and 72 h after transfection. Transfection efficiency was tested at 24 h with GFP-labelled vector (Figure 5 Bottom).

The human gastric cancer cell lines AGS (derived from the stomach tissue of a 54-year-old female patient with gastric adenocarcinoma ) and SGC-7901 (isolated from the metastasis of untreated gastric adenocarcinoma of a 56-year-old female patient) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and grown at 37 °C in Dulbecco’s Modified Eagle’s Medium (DMEM medium, Hyclone, Logan, Utah, USA, SH30081.02) supplemented with 10% fetal bovine serum (FBS, Gibco, California, USA, 10,091,148), 2 mM L-glutamine (Invitrogen, Carlsbad, California USA, 21,051,040),1% penicillin (100 units/ml) and streptomycin (100 µg/ml) (Invitrogen, Carlsbad, California USA, 15,140,122).
For stable overexpression of CHFR in AGS and SGC-7901 cells, the CHFR cDNA was amplified by PCR and subcloned into the LV-13 (pLenti-EF1a-LUC-F2A-Puro-CMV) vector for lentivirus package (GenePharma). AGS and SGC-7901 cells were infected with the concentrated virus with CHFR overexpression vector or empty vector. Subsequently, cells were treated with 2 µg/ml puromycin for 2 weeks to select cells with stable expression of CHFR, and the expression efficiency was validated by western blot analysis.