Identifying chemopreventive agents for obesity-associated cancers using an efficient, 3D high-throughput transformation assay
Identifying chemopreventive agents for obesity-associated cancers using an efficient, 3D high-throughput transformation assay
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DOI：
10.1038/s41598-019-46531-y
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发表时间：
2019-07-16
影响因子：
4.6
通讯作者：
Bernard, Jamie J.
中科院分区：
文献类型：
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作者：
Benham, Vanessa;Bullard, Blair;Bernard, Jamie J.
肥胖与大约40%的癌症诊断有关，但目前还没有有效的预防策略，这说明了化学预防的必要性。我们之前证明了脂肪组织中的成纤维细胞生长因子2 （FGF2）刺激恶性转化，这是通过软琼脂生长来测量的，这是体外转化实验的金标准。由于软琼脂实验不适合高通量筛选（HTS），我们开发了一种在超低附着条件下使用3D生长的新方法，作为琼脂生长的替代方法，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，在类似于软琼脂生长的超低附着条件下刺激生长。这种转化HTS鉴定出胰岛素生长因子1受体（IGF1R）抑制剂picropodophylin和HMG-CoA还原酶抑制剂氟伐他汀作为潜在的化学预防剂。这些化合物在软琼脂中使用两种非致瘤性细胞系验证了其功效。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。这种新颖的3D HTS平台快速、稳健，具有识别肥胖相关癌症预防药物的潜力。
Obesity is associated with similar to 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.