Li Qun, Chen Chaoyu, Kapadia Amit, Zhou Qiong, Harper Mary Kay, Schaack Jerome, LaBarbera Daniel V
Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Denver Anschutz Medial Center, Aurora, CO 80045, USA.
J Biomol Screen. 2011 Feb;16(2):141-54. doi: 10.1177/1087057110392995.
Despite advancements in therapies developed for the treatment of cancer, patient prognosis and mortality rates have improved minimally, and metastasis remains the primary cause of cancer mortality worldwide. An underlying mechanism promoting metastasis in many types of cancer is epithelial-mesenchymal transition (EMT). Here the authors report a novel 3D model of EMT and metastatic breast cancer suitable for high-throughput screening (HTS) drug discovery. The primary assay incorporates the expression of the prognostic biomarker vimentin, as a luciferase reporter of EMT, in basil-like/triple-negative MDA-MB-231 breast carcinoma spheroids. Using this model, the authors developed a number of known antitumor agents as control modulators of EMT. U0126, PKC412, PF2341066, dasatinib, and axitinib downregulated vimentin expression by 70% to 90% as compared to untreated spheroids. Counterassays were developed to measure spheroid viability and the invasive potential of MDA-MB-231 spheroids after small-molecule treatment and used to confirm hits from primary screening. Finally, the authors conducted a pilot screen to validate this model for HTS using a purified library of marine secondary metabolites. From 230 compounds screened, they obtained a Z' score of 0.64, indicative of an excellent assay, and confirmed 4 hits, including isonaamidine B, papuamine, mycalolide E, and jaspamide. This HTS model demonstrates the potential to identify small-molecule modulators of EMT that could be used to discover novel antimetastatic agents for the treatment of cancer.
尽管在癌症治疗方面取得了进展，但患者的预后和死亡率改善甚微，转移仍然是全球癌症死亡的主要原因。上皮-间质转化（EMT）是多种癌症中促进转移的潜在机制。在此，作者报告了一种适用于高通量筛选（HTS）药物发现的EMT和转移性乳腺癌的新型三维模型。主要检测方法是在基底样/三阴性MDA-MB-231乳腺癌球体中，将预后生物标志物波形蛋白的表达作为EMT的荧光素酶报告基因。利用该模型，作者开发了多种已知的抗肿瘤药物作为EMT的对照调节剂。与未处理的球体相比，U0126、PKC412、PF2341066、达沙替尼和阿西替尼使波形蛋白表达下调了70%至90%。开发了反检测方法来测量小分子处理后MDA-MB-231球体的活力和侵袭潜力，并用于确认初筛中的阳性结果。最后，作者使用纯化的海洋次生代谢产物文库进行了一次初步筛选，以验证该模型用于HTS的有效性。从筛选的230种化合物中，他们获得了0.64的Z'分数，表明该检测方法非常出色，并确认了4个阳性结果，包括异那米定B、帕普胺、麦考内酯E和茉莉酰胺。这种HTS模型显示了识别EMT小分子调节剂的潜力，这些调节剂可用于发现治疗癌症的新型抗转移药物。
