Department of Pediatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Curr Med Sci. 2020 Apr;40(2):354-362. doi: 10.1007/s11596-020-2181-5. Epub 2020 Apr 26.
Imatinib mesylate (IM) is the first-line treatment for Philadelphia (Ph) chromosomal positive leukemia by inhibiting phosphorylation of substrates via binding to the ABL kinase domain. Because of the drug resistance, side effects and the high cost of IM, it is necessary to find anti-cancer drugs with relatively low toxicity and cost, and enhanced efficacy, such as traditional Chinese medicines (TCMs). As one of TCMs, Huai Qi Huang (HQH) was chosen to treat BV173 and K562 cells. Various concentrations of HQH were added to cells for 24-72 h. Co-treatment of HQH and trametinib, an MEK inhibitor, was used to verify the synergistic effects on cell viability and apoptosis. Knockdown and overexpression of mitogen-activated protein kinase kinase 4 (MEK4) were implemented to demonstrate the role of MEK in cell apoptosis. Cell viability and apoptosis were measured by cell counting kit-8 assay (CCK8) and flow cytometry, respectively. Western blotting and real-time quantitative PCR (RT-qPCR) were used to assess protein and mRNA expression levels, respectively. The results showed that HQH inhibited survival and promoted apoptosis of BV173 and K562 cells in a dose-dependent manner, accompanied with down-regulation of PRKCH mRNA as well as CRAF, MEK4, phospho-ERK (pERK) and BCL2 proteins, and up-regulation of cleaved caspase3 protein. Co-treatment of HQH and trametinib had a synergistic effect on inhibiting survival and promoting apoptosis. MEK4 knockdown increased apoptosis, and had a synergistic effect with HQH. In contrast, MEK4 overexpression decreased apoptosis, and had the opposite effect with HQH. Collectively, the results of this study may identify a therapeutic mechanism of HQH on promoting apoptosis, and provide a potential option for treatment of Ph leukemia.
甲磺酸伊马替尼（IM）通过与 ABL 激酶结构域结合抑制底物磷酸化，是治疗费城染色体阳性白血病的一线药物。由于 IM 的耐药性、副作用和高昂的成本，有必要寻找毒性和成本相对较低、疗效增强的抗癌药物，如中药（TCM）。作为 TCM 之一，槐杞黄（HQH）被用于治疗 BV173 和 K562 细胞。将不同浓度的 HQH 加入细胞中 24-72 小时。用 MEK 抑制剂 trametinib 与 HQH 共同处理，以验证对细胞活力和凋亡的协同作用。敲低和过表达丝裂原活化蛋白激酶激酶 4（MEK4），以证明 MEK 在细胞凋亡中的作用。通过细胞计数试剂盒-8 测定法（CCK8）和流式细胞术分别测量细胞活力和细胞凋亡。Western blot 和实时定量 PCR（RT-qPCR）分别用于评估蛋白和 mRNA 表达水平。结果表明，HQH 呈剂量依赖性抑制 BV173 和 K562 细胞的存活并促进其凋亡，同时下调 PRKCH mRNA 以及 CRAF、MEK4、磷酸化 ERK（pERK）和 BCL2 蛋白，并上调 cleaved caspase3 蛋白。HQH 与 trametinib 联合处理对抑制存活和促进凋亡具有协同作用。MEK4 敲低增加凋亡，并与 HQH 具有协同作用。相反，MEK4 过表达减少凋亡，并与 HQH 具有相反的作用。总之，这项研究的结果可能确定了 HQH 促进凋亡的治疗机制，并为治疗 Ph 白血病提供了一种潜在的选择。
