JaponiconeA induces apoptosis of bortezomib-sensitive and -resistant myeloma cells in vitro and in vivo by targeting IKK.
JaponiconeA induces apoptosis of bortezomib-sensitive and -resistant myeloma cells in vitro and in vivo by targeting IKK.
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DOI：
10.20892/j.issn.2095-3941.2020.0473
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发表时间：
2021-09-28
影响因子：
5.5
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中科院分区：
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多发性骨髓瘤（MM）仍然无法治愈，复发率高。因此，迫切需要新的治疗药物来改善预后。从旋覆花中分离得到的天然产物JaponiconeA（JA）具有良好的抗MM活性。因此，应进行全面的研究，以确定JA抗MM作用的体外和体内机制。CCK 8法和流式细胞仪检测JA对MM细胞增殖、凋亡和细胞周期的影响。使用皮下异种移植小鼠模型进行体内实验。我们还使用RNA-seq和c-Map数据库鉴定了JA的可能靶标和机制，并使用CETSA、DARTS和拯救实验鉴定了JA在硼替佐米敏感和耐药MM细胞系中的特异性靶标。此外，JA和硼替佐米单独或一起使用，以表征其可能的协同效应。JA在体外可抑制MM细胞增殖、诱导细胞凋亡和G2/M期阻滞，并选择性杀伤原代CD 138 + MM细胞。在体内，JA也表现出很强的抗肿瘤作用，没有观察到毒性。此外，JA与硼替佐米联合使用时表现出协同作用，并增强硼替佐米在硼替佐米耐药细胞中的抗肿瘤作用。CETSA和DARTS证实JA与NF-κB抑制剂激酶β（IKKβ）直接结合，过表达IKKβ或敲低IκBα可部分挽救JA诱导的细胞凋亡。JA对MM具有较强的抗肿瘤作用，可使骨髓瘤细胞对硼替佐米敏感，并通过抑制IKKβ克服NF-κ B诱导的耐药，为MM患者提供了一种新的治疗策略。
Multiple myeloma (MM) remains incurable with high rates of relapse. New therapeutic drugs are therefore urgently needed to improve the prognosis. JaponiconeA (JA), a natural product isolated from Inula japonica Thunb, has shown good anti-MM potential. A comprehensive study should therefore be conducted to identify both the in vitro and in vivo mechanisms of the anti-MM effects of JA. CCK8 assays and flow cytometry were used to detect the proliferation, apoptosis, and cell cycle of MM cell lines when treated with JA. In vivo experiments were conducted using subcutaneous xenograft mouse models. We also identified possible targets and the mechanism of JA using RNA-seq and c-Map databases, and identified the specific targets of JA in bortezomib-sensitive and -resistant MM cell lines using CETSA, DARTS, and rescue experiments. Furthermore, JA and bortezomib were used separately or together to characterize their possible synergistic effects. In vitro, JA inhibited proliferation, and induced apoptosis and G2/M phase arrest in MM cell lines, and selectively killed primary CD138+ MM cells. In vivo, JA also demonstrated a strong anti-tumor effect with no observable toxicity. In addition, JA showed synergetic effects in combination with bortezomib, and enhanced the anti-tumor effect of bortezomib in bortezomib-resistant cells. CETSA and DARTS confirmed direct binding of JA to NF-κB inhibitor kinase beta (IKKβ), and overexpression of IKKβ or knockdown of IκBα partially rescued the apoptosis induced by JA. JA exhibited strong anti-tumor effects in MM. It sensitized myeloma cells to bortezomib and overcame NF-κB-induced drug resistance by inhibiting IKKβ, providing a new treatment strategy for MM patients.