MCF-7 human mammary adenocarcinoma cells exhibit augmented responses to human insulin on a collagen IV surface
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Human mammary cell lines are extensively used for preclinical safety assessment of insulin analogs. However, it is essentially unknown how mitogenic responses can be optimized in mammary cell-based systems. We developed an insulin mitogenicity assay in MCF-7 human mammary adenocarcinoma cells, under low serum (0.1% FCS) and phenol red-free conditions, with 3H thymidine incorporation as endpoint. Based on EC50 values determined from 10-fold dilution series, beta-estradiol was the most potent mitogen, followed by human IGF-1, human AspB10 insulin and native human insulin. AspB10 insulin was significantly more mitogenic than native insulin, validating the ability of the assay to identify hypermitogenic human insulin analogs. With MCF-7 cells on a collagen IV surface, the ranking of mitogens was maintained, but fold mitogenic responses and dynamic range and steepness of dose-response curves were increased. Also, PI3K pathway activation by insulin was enhanced on a collagen IV surface. This study provided the first determination and ranking of the mitogenic potencies of standard reference compounds in an optimized MCF-7 assay. The optimized MCF-7 assay described here is of relevance for in vitro toxicological testing and carcinogenicity safety assessment of new insulin compounds.
Originalsprog | Engelsk |
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Tidsskrift | Journal of Applied Toxicology |
Vol/bind | 29 |
Udgave nummer | 6 |
Sider (fra-til) | 470-7 |
ISSN | 0260-437X |
DOI | |
Status | Udgivet - 2009 |
Bibliografisk note
Copyright 2009 John Wiley & Sons, Ltd.
- Det tidligere Farmaceutiske Fakultet
Forskningsområder
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