Ma Yihong, Kurtyka Courtney A, Boyapalle Sandhya, Sung Shen-Shu, Lawrence Harshani, Guida Wayne, Cress W Douglas
Molecular Oncology Program, H Lee Moffitt Cancer Center and Research Institute, Tampa, Florida 33612, USA.
Cancer Res. 2008 Aug 1;68(15):6292-9. doi: 10.1158/0008-5472.CAN-08-0121.
HLM006474 was identified using a computer-based virtual screen and the known crystal structure of the DNA-bound E2F4/DP2 heterodimer. Treatment of multiple cell lines with HLM006474 resulted in the loss of intracellular E2F4 DNA-binding activity as measured by electrophoretic mobility shift assay within hours. Overnight exposure to HLM006474 resulted in down-regulation of total E2F4 protein as well as known E2F targets. The effects of HLM006474 treatment on different cell lines varied but included a reduction in cell proliferation and an increase in apoptosis. HLM006474 induced apoptosis in a manner distinct from cisplatin and doxorubicin. E2F4-null mouse embryonic fibroblasts were less sensitive than wild-type counterparts to the apoptosis-inducing activity of the compound, revealing its biological specificity. A375 cells were extremely sensitive to the apoptosis-inducing activity of the compound in two-dimensional culture, and HLM006474 was a potent inhibitor of melanocytes proliferation and subsequent invasion in a three-dimensional tissue culture model system. Together, these results suggest that interference with E2F activity using small molecules may have clinical application in cancer therapy.
HLM006474是通过基于计算机的虚拟筛选以及与DNA结合的E2F4/DP2异二聚体的已知晶体结构鉴定出来的。用HLM006474处理多种细胞系，数小时内通过电泳迁移率变动分析测定，细胞内E2F4 DNA结合活性丧失。过夜暴露于HLM006474导致总E2F4蛋白以及已知的E2F靶标下调。HLM006474处理对不同细胞系的影响各不相同，但包括细胞增殖减少和凋亡增加。HLM006474诱导凋亡的方式与顺铂和阿霉素不同。E2F4基因敲除的小鼠胚胎成纤维细胞对该化合物的凋亡诱导活性不如野生型细胞敏感，揭示了其生物学特异性。A375细胞在二维培养中对该化合物的凋亡诱导活性极其敏感，并且在三维组织培养模型系统中，HLM006474是黑素细胞增殖和随后侵袭的有效抑制剂。总之，这些结果表明，使用小分子干扰E2F活性可能在癌症治疗中具有临床应用价值。
