Classifying the adverse mitogenic mode of action of insulin analogues using a novel mechanism-based genetically engineered human breast cancer cell panel

• Published in: Archives of toxicology Vol. 88; no. 4; pp. 953 - 966
• Main Authors: ter Braak, B., Siezen, C. L. E., Kannegieter, N., Koedoot, E., van de Water, B., van der Laan, J. W.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.04.2014; Springer Nature B.V
• Subjects: Antigens, CD - genetics; Antigens, CD - metabolism; Biomedical and Life Sciences; Biomedicine; Biotransformation; Breast cancer; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cell Proliferation - drug effects; Classification; Dose-Response Relationship, Drug; Environmental Health; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases - metabolism; Female; Genetic engineering; Genetic Engineering - methods; Humans; Hypoglycemic Agents - metabolism; Hypoglycemic Agents - toxicity; In Vitro Systems; Insulin; Insulin - analogs & derivatives; Insulin - metabolism; Insulin - toxicity; Insulin Glargine - toxicity; MCF-7 Cells; Occupational Medicine/Industrial Medicine; Pharmacology/Toxicology; Proto-Oncogene Proteins c-akt - metabolism; Receptor, Insulin - agonists; Receptor, Insulin - genetics; Receptor, Insulin - metabolism; Receptors, Somatomedin - agonists; Receptors, Somatomedin - genetics; Receptors, Somatomedin - metabolism; RNA Interference; Signal Transduction - drug effects; Time Factors; Toxicology; Transfection; Insulin glargine; Mitogenic; Human breast cancer; IGF1R
• ISSN: 0340-5761; 1432-0738
• DOI: 10.1007/s00204-014-1201-2

Insulin analogues are widely used in clinical practice. Modifications on the insulin molecular structure can affect the affinity and activation towards two closely related receptor tyrosine kinases: the insulin receptor (INSR) and the insulin-like growth factor 1 receptor (IGF1R). A switch towards higher IGF1R affinity is likely to emphasize mitogenesis rather than glucose metabolism. Relevant well-validated experimental tools to address the insulin analogue activation of either INSR or IGF1R are missing. We have established a panel of human MCF-7 breast cancer cell lines either ectopically expressing the INSR (A or B isoform) in conjunction with a stable knockdown of the IGF1R or ectopically expressing the IGF1R in conjunction with a stable knockdown of the INSR. In these cell lines, we systematically evaluated the INSR and IGF1R receptor activation and downstream mitogenic signalling of all major clinical relevant insulin analogues in comparison with insulin and IGF1R. While most insulin analogues primarily activated the INSR, the mitogenic activation pattern of glargine was highly similar to IGF1 and insulin AspB10, known to bind IGF1R and induce carcinogenesis. Yet, in a long-term proliferation assay, the proliferative effect of glargine was not much different from regular insulin or other insulin analogues. This was caused by the rapid enzymatic conversion into its two metabolic active metabolites M1 and M2, with reduced mitogenic signalling through the IGF1R. In summary, based on our new cell models, we identified a similar mitogenic potency of insulin glargine and AspB10. However, rapid enzymatic conversion of glargine precludes a sustained activation of the IGF1R signalling pathway.