Carrasco-Garcia Estefania, Martinez-Lacaci Isabel, Mayor-López Leticia, Tristante Elena, Carballo-Santana Mar, García-Morales Pilar, Ventero Martin Maria Paz, Fuentes-Baile Maria, Rodriguez-Lescure Álvaro, Saceda Miguel
Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, 03202 Alicante, Spain.
División de Neurooncología, Instituto Biodonostia, 20014 San Sebastián (Guipuzkoa), Spain.
Cells. 2018 Sep 6;7(9):131. doi: 10.3390/cells7090131.
Glioblastomas are highly resistant to radiation and chemotherapy. Currently, there are no effective therapies for this type of tumor. Signaling mechanisms initiated by PDGFR and IGF-1R are important in glioblastoma, and inhibition of the signal transduction pathways initiated by these receptors could be a useful alternative strategy for glioblastoma treatment. We have studied the effects of the PDGFR inhibitor JNJ-10198409 (JNJ) and the IGF-1R inhibitor picropodophyllin (PPP) in glioblastoma cell lines as well as in primary cultures derived from patients affected by this type of tumor. JNJ and PPP treatment blocked PDGFR and IGF-1R signaling respectively and reduced Akt and Erk 1/2 phosphorylation. Both inhibitors diminished cell proliferation, inducing a G2/M block of the cell cycle. Cell death induced by JNJ was caspase-dependent, Annexin-V positive and caused PARP cleavage, especially in T98 cells, suggesting an apoptotic mechanism. However, cell death induced by PPP was not completely inhibited by caspase inhibitors in all cell lines apart from LN-229 cells, indicating a caspase-independent mechanism. Several inhibitors targeted against different cell death pathways could not block this caspase-independent component, which may be a non-programmed necrotic mechanism. Apoptotic arrays performed in T98 and LN-229 cells upon JNJ and PPP treatment revealed that procaspase 3 levels were augmented by both drugs in T98 cells and only by JNJ in LN229-cells. Furthermore, XIAP and survivin levels were much higher in LN-229 cells than in T98 cells, revealing that LN-229 cells are more susceptible to undergo caspase-independent cell death mechanisms. JNJ and PPP combination was more effective than each treatment alone.
胶质母细胞瘤对放疗和化疗具有高度抗性。目前,对于这类肿瘤尚无有效的治疗方法。由血小板衍生生长因子受体(PDGFR)和胰岛素样生长因子-1受体(IGF-1R)启动的信号传导机制在胶质母细胞瘤中很重要,抑制这些受体启动的信号转导途径可能是胶质母细胞瘤治疗的一种有用的替代策略。我们已经研究了PDGFR抑制剂JNJ-10198409(JNJ)和IGF-1R抑制剂鬼臼苦素(PPP)对胶质母细胞瘤细胞系以及源自这类肿瘤患者的原代培养物的影响。JNJ和PPP处理分别阻断了PDGFR和IGF-1R信号传导,并降低了Akt和细胞外信号调节激酶1/2(Erk 1/2)的磷酸化。两种抑制剂均减少了细胞增殖,诱导细胞周期的G2/M期阻滞。JNJ诱导的细胞死亡是半胱天冬酶依赖性的,膜联蛋白-V阳性并导致聚腺苷二磷酸核糖聚合酶(PARP)裂解,尤其是在T98细胞中,提示存在凋亡机制。然而,除LN-229细胞外,PPP诱导的细胞死亡在所有细胞系中均未被半胱天冬酶抑制剂完全抑制,表明存在半胱天冬酶非依赖性机制。针对不同细胞死亡途径的几种抑制剂均无法阻断这种半胱天冬酶非依赖性成分,这可能是一种非程序性坏死机制。在JNJ和PPP处理后对T98和LN-229细胞进行的凋亡阵列分析显示,两种药物均使T98细胞中的半胱天冬酶原-3水平升高,而在LN229细胞中仅JNJ使其升高。此外,X连锁凋亡抑制蛋白(XIAP)和生存素水平在LN-229细胞中比在T98细胞中高得多,这表明LN-229细胞更容易发生半胱天冬酶非依赖性细胞死亡机制。JNJ和PPP联合使用比单独使用每种治疗更有效。