Expression data from human carcinoma (MCF7) derived cells that have been exposed to insulin analogues

Insulin analogues are designed to improve the pharmacokinetic parameters compared to regular human insulin. This provides a sustained control of blood glucose levels in diabetic patients. All novel insulin analogues are tested for their mitogenic side effects, however these assays do not take into account the molecular mode-of-action of different insulin analogues. Insulin analogues can bind the insulin receptor (INSR) and the insulin-like growth factor-1 receptor (IGF1R) with different affinities and consequently will activate different downstream signaling pathways. Here we used a panel of MCF7 human breast cancer cell lines that selectively express either one of the isoforms of the INSR (IRA or IRB) or the IGF1R. We sought to study the role of the different receptors (IRA, IRB and IGF1R) in the mitogenic signaling of insulin-like molecules (including insulin, glargine, X10 (or AspB10) and IGF1). MCF7 IRA, MCF7 IRB or MCF7 IGF1R cells (as described in Arch Toxicol. 2014 Apr;88(4):953-66. doi: 10.1007/s00204-014-1201-2. Epub 2014 Jan 25.) were cultured in RPMI supplemented with 5% (v/v) CDFBS (Hyclone) and used for experiments. Cells have been exposed for 1 or 6 hours to 10 nM of the indicated insulin-like molecule. As a control sample a vehicle stimulation was performed that contained everything except the active compound.

胰岛素类似物（insulin analogues）的设计初衷是相较于常规人胰岛素优化其药代动力学参数，从而实现糖尿病患者血糖水平的持续管控。所有新型胰岛素类似物均需检测其促有丝分裂副作用，但现有检测手段并未考量不同胰岛素类似物的分子作用模式。胰岛素类似物可与胰岛素受体（insulin receptor, INSR）及胰岛素样生长因子-1受体（insulin-like growth factor-1 receptor, IGF1R）以不同亲和力结合，进而激活不同的下游信号通路。
本研究采用一组可选择性表达胰岛素受体两种亚型（IRA或IRB）或IGF1R的人乳腺癌MCF7细胞系，旨在探究不同受体（IRA、IRB及IGF1R）在胰岛素样分子（包括胰岛素、甘精胰岛素、X10（或称AspB10）及IGF1）的促有丝分裂信号通路中的作用。MCF7 IRA、MCF7 IRB及MCF7 IGF1R细胞（详见Arch Toxicol. 2014 Apr;88(4):953-66. doi: 10.1007/s00204-014-1201-2. Epub 2014 Jan 25.）均采用添加5%（v/v）CDFBS（Hyclone）的RPMI培养基培养，用于后续实验。将细胞与指定浓度为10 nM的胰岛素样分子孵育1小时或6小时；设置溶剂刺激对照组，该对照组除不含活性化合物外，其余成分与实验组完全一致。