MALAT1 Overexpression and Silencing in Osteosarcoma

The procedure was carried out as previously described [63 (link)]. MNNG/HOS cells at 60~80% confluence were then selected for cell transfection. Plasmids were transfected into osteosarcoma cells by using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's protocol. The sequence of MALAT1 siRNAs were as follows as ascribed before [21 (link), 64 (link)]: 1# MALAT1 siRNA, 5’-GGCAAUGUUUUACACUAUUTT-3’; 2# MALAT1 siRNA, 5’-CACAGGGAAAGCGAGTGGTTGGTAA-3’; 3# MALAT1 siRNA, 5’-CACAGGGAAAGCGAGUGGUUGGU-3’; non-specific siRNA 5’-UUCUCCGAACGUGUCACGUTT-3’. MALAT1-siRNA and non-specific siRNA were purchased from Invitrogen (USA). To construct MALAT1 stable over-expression cell lines for further animal study, the stable transfected pcDNA3.1-MALAT1-wt and corresponding control (pcDNA3.1) cells were selected by the culture medium containing 0.4 mg/ml Geneticin (G418, Invitrogen). After 6 weeks, G418-resistant cell clones were established to construct MALAT1 stable over-expression cell lines.