PDF(Pubmed)
Abstract：
Adjuvant treatment for Glioblastoma Grade 4 with Temozolomide (TMZ) inevitably fails due to therapeutic resistance, necessitating new approaches. Apoptosis induction in GB cells is inefficient, due to an excess of anti-apoptotic XPO1/Bcl-2-family proteins. We assessed TMZ, Methotrexate (MTX), and Cytarabine (Ara-C) (apoptosis inducers) combined with XPO1/Bcl-2/Mcl-1-inhibitors (apoptosis rescue) in GB cell lines and primary GB stem-like cells (GSCs). Using CellTiter-Glo® and Caspase-3 activity assays, we generated dose-response curves and analyzed the gene and protein regulation of anti-apoptotic proteins via PCR and Western blots. Optimal drug combinations were examined for their impact on the cell cycle and apoptosis induction via FACS analysis, paralleled by the assessment of potential toxicity in healthy mouse brain slices. Ara-C and MTX proved to be 150- to 10,000-fold more potent in inducing apoptosis than TMZ. In response to inhibitors Eltanexor (XPO1; E), Venetoclax (Bcl-2; V), and A1210477 (Mcl-1; A), genes encoding for the corresponding proteins were upregulated in a compensatory manner. TMZ, MTX, and Ara-C combined with E, V, and A evidenced highly lethal effects when combined. As no significant cell death induction in mouse brain slices was observed, we conclude that this drug combination is effective in vitro and expected to have low side effects in vivo.
摘要：
替莫唑胺（TMZ）辅助治疗4级胶质母细胞瘤不可避免地由于治疗耐药性而失败。需要新的方法。在GB细胞中诱导凋亡是无效的，由于过量的抗凋亡XPO1/Bcl-2家族蛋白。我们评估了TMZ,甲氨蝶呤(MTX),和阿糖胞苷（Ara-C）（凋亡诱导剂）联合XPO1/Bcl-2/Mcl-1抑制剂（凋亡拯救）在GB细胞系和原代GB干细胞样细胞（GSC）中。使用CellTiter-Glo®和Caspase-3活性测定，我们产生剂量-反应曲线,并通过PCR和Western印迹分析抗凋亡蛋白的基因和蛋白调控.通过FACS分析检查了最佳药物组合对细胞周期和凋亡诱导的影响。同时评估健康小鼠脑切片的潜在毒性。事实证明，Ara-C和MTX在诱导凋亡方面的效力比TMZ高150至10,000倍。在对抑制剂Eltanexor（XPO1;E）的反应中，维奈托克（Bcl-2；V），和A1210477(Mcl-1;A)，编码相应蛋白质的基因以代偿方式上调。TMZ,MTX,Ara-C与E结合,V,和被证明的高度致命的影响时，结合。由于在小鼠脑切片中没有观察到显著的细胞死亡诱导,我们得出的结论是，这种药物组合在体外是有效的，并且在体内具有低副作用。
