School of Pharmacy, Health Science Center, Xi'an Jiaotong University, No. 76, Yanta Weststreet, #54, Xi'an, Shaanxi Province 710061, PR China.
School of Pharmacy, Health Science Center, Xi'an Jiaotong University, No. 76, Yanta Weststreet, #54, Xi'an, Shaanxi Province 710061, PR China.
Biomed Pharmacother. 2018 Jul;103:118-126. doi: 10.1016/j.biopha.2018.03.174. Epub 2018 Apr 24.
Cervical carcinoma remains the second most common malignancy with a high mortality rate among women worldwide. TAD-1822-7-F2 (F2) and TAD-1822-7-F5 (F5) are novel compounds synthesized on the chemical structure of taspine derivatives, and show an effective suppression for HeLa cells. Our study aims to confirm the potential targets of F2 and F5, and investigate the underlying mechanism of the inhibitory effect on HeLa cells. In this study, Real Time Cell Analysis and crystal violet staining assay were conducted to investigate the effect of F2 and F5 on HeLa cells proliferation. And the analytical methods of surface plasmon resonance and quartz crystal microbalance were established and employed to study the interaction between F2 and F5 and potential target protein JAK2, suggesting that both compounds have strong interaction with the JAK2 protein. Western blot analysis, immunofluorescence staining study and PCR was conducted to investigate the molecules of JAK/Stat signaling pathway. Interestingly, F2 and F5 showed diverse regulation for signaling molecules because of their different chemical structure. F2 increased the expression of JAK2 and downregulated the level of P-JAK1 and P-JAK2, and decreased P-Stat3 (Ser727). While F5 could increase the expression of JAK2 and naturally decrease the phosphorylation of JAK1 and Tyk2, and decreased the expression of P-Stat6. Moreover, F2 and F5 showed the same downregulation on the P-Stat3 (Tyr705). Therefore, F2 and F5 could target the JAK2 protein and prevent the phosphorylation of JAKs to suppress the phosphorylation of the downstream effector Stats, which suggested that F2 and F5 have great potential to be the inhibitors of the JAK/Stat signaling pathway.
宫颈癌仍然是全球女性中死亡率第二高的常见恶性肿瘤。TAD-1822-7-F2（F2）和 TAD-1822-7-F5（F5）是在 taspine 衍生物化学结构基础上合成的新型化合物，对 HeLa 细胞具有有效的抑制作用。本研究旨在确认 F2 和 F5 的潜在靶点，并研究其对 HeLa 细胞抑制作用的潜在机制。在这项研究中，采用实时细胞分析和结晶紫染色实验来研究 F2 和 F5 对 HeLa 细胞增殖的影响。并建立了表面等离子体共振和石英晶体微天平的分析方法，用于研究 F2 和 F5 与潜在靶蛋白 JAK2 的相互作用，表明这两种化合物与 JAK2 蛋白具有很强的相互作用。通过 Western blot 分析、免疫荧光染色研究和 PCR 研究 JAK/Stat 信号通路的分子。有趣的是，由于其不同的化学结构，F2 和 F5 对信号分子表现出不同的调节作用。F2 增加了 JAK2 的表达，下调了 P-JAK1 和 P-JAK2 的水平，并降低了 P-Stat3（Ser727）的水平。而 F5 可以增加 JAK2 的表达，并自然降低 JAK1 和 Tyk2 的磷酸化水平，降低 P-Stat6 的表达。此外，F2 和 F5 对 P-Stat3（Tyr705）也表现出相同的下调作用。因此，F2 和 F5 可以靶向 JAK2 蛋白，阻止 JAKs 的磷酸化，从而抑制下游效应器 Stats 的磷酸化，这表明 F2 和 F5 具有很大的潜力成为 JAK/Stat 信号通路的抑制剂。
