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首页> 外文期刊>Journal of applied toxicology >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.

机译:MCF-7人乳腺腺癌细胞在胶原IV表面上对人胰岛素的反应增强。

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摘要

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.
机译:人乳腺细胞系被广泛用于胰岛素类似物的临床前安全性评估。但是,基本上未知如何在基于乳腺细胞的系统中优化有丝分裂反应。我们在低血清(0.1%FCS)和无酚红的条件下,以3H胸苷掺入为终点,在MCF-7人乳腺腺癌细胞中开发了一种胰岛素促有丝分裂性测定法。根据从10倍稀释系列确定的EC50值,β-雌二醇是最有效的促分裂原,其次是人IGF-1,人AspB10胰岛素和天然人胰岛素。 AspB10胰岛素比天然胰岛素有更多的促有丝分裂作用,从而证实了该方法能够鉴定出有丝分裂原性的人胰岛素类似物。在胶原蛋白IV表面上使用MCF-7细胞,可维持有丝分裂原的排名,但有丝分裂反应的倍数以及剂量反应曲线的动态范围和陡度增加。同样,胰岛素在胶原蛋白IV表面上的PI3K途径活化得到增强。这项研究提供了在优化的MCF-7分析中标准参考化合物的促有丝分裂能力的首次测定和排名。此处描述的优化的MCF-7测定与新胰岛素化合物的体外毒理学测试和致癌性安全性评估相关。

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