MCF-7 human mammary adenocarcinoma cells exhibit augmented responses to human insulin on a collagen IV surface
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MCF-7 human mammary adenocarcinoma cells exhibit augmented responses to human insulin on a collagen IV surface. / Listov-Saabye, Nicolai; Jensen, Marianne Blirup; Kiehr, Benedicte; Hansen, Erik Wind; Svendsen, Jette E; Lundby, Anders; Holm, Gitte-Mai Nelander; Oleksiewicz, Martin B.
In: Journal of Applied Toxicology, Vol. 29, No. 6, 2009, p. 470-7.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - MCF-7 human mammary adenocarcinoma cells exhibit augmented responses to human insulin on a collagen IV surface
AU - Listov-Saabye, Nicolai
AU - Jensen, Marianne Blirup
AU - Kiehr, Benedicte
AU - Hansen, Erik Wind
AU - Svendsen, Jette E
AU - Lundby, Anders
AU - Holm, Gitte-Mai Nelander
AU - Oleksiewicz, Martin B
N1 - Copyright 2009 John Wiley & Sons, Ltd.
PY - 2009
Y1 - 2009
N2 - 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.
AB - 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.
KW - Former Faculty of Pharmaceutical Sciences
U2 - 10.1002/jat.1428
DO - 10.1002/jat.1428
M3 - Journal article
C2 - 19338014
VL - 29
SP - 470
EP - 477
JO - Journal of Applied Toxicology
JF - Journal of Applied Toxicology
SN - 0260-437X
IS - 6
ER -
ID: 14774425