Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI, 48824, USA.
Division of Dermatology, Department of Medicine, College of Human Medicine, Michigan State University, East Lansing, MI, 48824, USA.
Sci Rep. 2019 Jul 16;9(1):10278. doi: 10.1038/s41598-019-46531-y.
Obesity is associated with ~40% of cancer diagnoses but there are currently no effective preventive strategies, illustrating a need for chemoprevention. We previously demonstrated that fibroblast growth factor 2 (FGF2) from adipose tissue stimulates malignant transformation, as measured by growth in soft agar, the gold-standard in vitro transformation assay. Because the soft agar assay is unsuitable for high throughput screens (HTS), we developed a novel method using 3D growth in ultra-low attachment conditions as an alternative to growth in agar to discover compounds that inhibit transformation. Treating non-tumorigenic, skin epithelial JB6 P cells with FGF2 stimulates growth in ultra-low attachment conditions analogous to growth in the soft agar. This transformation HTS identified picropodophyllin, an insulin growth factor 1 receptor (IGF1R) inhibitor, and fluvastatin, an HMG-CoA reductase inhibitor, as potential chemopreventive agents. These compounds were validated for efficacy using two non-tumorigenic cell lines in soft agar. Another IGF1R inhibitor and other statins were also tested and several were able to inhibit growth in soft agar. This novel 3D HTS platform is fast, robust and has the potential to identify agents for obesity-associated cancer prevention.
肥胖与约 40%的癌症诊断有关，但目前尚无有效的预防策略，这表明需要进行化学预防。我们之前的研究表明，脂肪组织中的成纤维细胞生长因子 2（FGF2）通过软琼脂中的生长来刺激恶性转化，软琼脂是体外转化测定的金标准。由于软琼脂测定不适用于高通量筛选（HTS），我们开发了一种新方法，使用超低附着条件下的 3D 生长作为替代软琼脂中的生长，以发现抑制转化的化合物。用 FGF2 处理非致瘤性皮肤上皮 JB6 P 细胞可刺激超低附着条件下的生长，类似于软琼脂中的生长。这种转化 HTS 鉴定出了 picropodophyllin，一种胰岛素样生长因子 1 受体（IGF1R）抑制剂，和 fluvastatin，一种 HMG-CoA 还原酶抑制剂，作为潜在的化学预防剂。使用软琼脂中的两种非致瘤性细胞系验证了这些化合物的功效。另一种 IGF1R 抑制剂和其他他汀类药物也进行了测试，其中几种能够抑制软琼脂中的生长。这种新的 3D HTS 平台快速、稳健，有可能识别出用于预防肥胖相关癌症的药物。
