Norris Cotton Cancer Center, Rubin Building Level 6, Dartmouth Medical School, Lebanon, NH 03756, USA.
Mol Cancer Ther. 2010 Apr;9(4):791-802. doi: 10.1158/1535-7163.MCT-10-0028. Epub 2010 Apr 6.
Chemotherapeutic agents modify intracellular signaling that culminates in the inhibition of Bcl-2 family members and initiates apoptosis. Inhibition of the extracellular signal-regulated kinase by PD98059 dramatically accelerates vinblastine-mediated apoptosis in ML-1 leukemia with cells dying in 4 hours from all phases of the cell cycle. Inhibition of protein synthesis by cycloheximide also markedly accelerated vinblastine-induced apoptosis, showing that the proteins required for this acute apoptosis are constitutively expressed. Vinblastine induced the rapid induction of Mcl-1 that was inhibited by PD98059 and cycloheximide. No change in Bcl-2 or Bcl-X was observed. We hypothesize that ML-1 cells use Mcl-1 for protection from the rapid vinblastine-induced apoptosis. This was confirmed by targeting Mcl-1 with short hairpin RNA. We also investigated the response of 13 other leukemia and lymphoma cell lines and cells from seven chronic lymphocytic leukemia patients. Four cell lines and all chronic lymphocytic leukemia cells were killed in 6 hours by vinblastine alone. Two additional cell lines were sensitized to vinblastine by PD98059, which suppressed Mcl-1. This acute apoptosis either alone or in combination with PD98059 required vinblastine-mediated activation of c-Jun-NH(2)-terminal kinase. PD98059 did not suppress Mcl-1 in other cell lines whereas sorafenib did, but this did not sensitize the cells to vinblastine, suggesting that the acute apoptosis varies depending on which Bcl-2 protein mediates protection. Most of the cell lines were sensitized to vinblastine by cycloheximide, suggesting that inhibition of a short-lived protein in addition to Mcl-1 can acutely sensitize cells. These results suggest several clinical strategies that might provide an effective therapy for selected patients. Mol Cancer Ther; 9(4); 791-802. (c)2010 AACR.
化疗药物会改变细胞内信号,最终抑制 Bcl-2 家族成员并引发细胞凋亡。通过 PD98059 抑制细胞外信号调节激酶可显著加速 ML-1 白血病细胞中长春碱介导的凋亡,所有细胞周期阶段的细胞在 4 小时内死亡。通过环己酰亚胺抑制蛋白质合成也显著加速长春碱诱导的凋亡,表明这种急性细胞凋亡所需的蛋白质是组成性表达的。长春碱诱导 Mcl-1 的快速诱导,而 Mcl-1 被 PD98059 和环己酰亚胺抑制。没有观察到 Bcl-2 或 Bcl-X 的变化。我们假设 ML-1 细胞使用 Mcl-1 来防止快速长春碱诱导的凋亡。通过短发夹 RNA 靶向 Mcl-1 证实了这一点。我们还研究了 13 种其他白血病和淋巴瘤细胞系以及 7 例慢性淋巴细胞白血病患者的细胞反应。单独使用长春碱,4 种细胞系和所有慢性淋巴细胞白血病细胞在 6 小时内被杀死。另外两种细胞系通过 PD98059 对长春碱敏感,PD98059 抑制了 Mcl-1。这种急性细胞凋亡无论是单独使用还是与 PD98059 联合使用,都需要长春碱介导的 c-Jun-NH(2)-末端激酶的激活。PD98059 不会抑制其他细胞系中的 Mcl-1,而索拉非尼会,但这不会使细胞对长春碱敏感,这表明急性细胞凋亡取决于哪种 Bcl-2 蛋白介导保护。大多数细胞系对长春碱敏感,提示除 Mcl-1 之外抑制一种短寿命蛋白也可以使细胞迅速敏感。这些结果表明了几种临床策略,这些策略可能为选定的患者提供有效的治疗方法。 Mol Cancer Ther; 9(4); 791-802. (c)2010 AACR。