Insuman basal

Next, an in vitro experiment was performed to reveal the transcriptomic effects of insulin analogue exposure on a human breast cancer cell line. MCF7 human breast cancer cells with an overexpression of the IGF1R and a stable knockdown of the IR (MCF7 IGF1R) were seeded at 60% confluence and starved for 2 days in 5% charcoal/dextran-treated FBS (CDFBS, Hyclone, USA) containing RPMI 1640 (Gibco, USA) medium. Cells were stimulated (with 10 nM compound) for 1 or 6 h after which RNA was isolated using NucleoSpin® miRNA isolation kit (Machery Nagel, Düren, Germany). Stimulations included: insulin NPH (Insuman Basal, Sanofi Aventis), insulin glargine (Lantus, Sanofi Aventis), insulin X10 (AspB10, Novo Nordisk), and IGF1 (Increlex, Ipsen). For more technical details about cell line generation and characterization or growth factor stimulation, please refer to our previous publication [7 ].

The treatments included: insulin NPH (Insuman Basal, Sanofi Aventis, Gouda, The Netherlands), insulin glargine (Lantus, Sanofi Aventis), insulin X10 (AspB10, Novo Nordisk, Alphen aan den Rijn, The Netherlands) and IGF1 (Increlex, Ipsen, Hoofddorp, The Netherlands). All compounds were dissolved in their original vehicle solutions: glargine (glycerol 0.2 mol/L, m-cresol 0.025 mol/L, ZnCl2 0.0002 mol/L adjusted to pH 4.0), insulin (glycerol 0.2 mol/L, NaH2PO4 0.00135 mol/L, phenol 0.0063 mol/L, m-cresol 0.0138 mol/L, ZnCl2 0.0001 mol/L adjusted to pH 7.4), X10 (glycerol 0.2 mol/L, phenol 0.0063 mol/L, m-cresol 0.0138 mol/L, ZnCl2 0.0001 mol/L adjusted to pH 7.4) and IGF1 (benzyl alcohol 0.083 mol/L, sodium chloride 0.1 mol/L, polysorbate 20 0.0016 mol/L, acetic acid 0.0072 mol/L, sodium acetate 0.05 mol/L adjusted to pH 5.4).