Autophagic cell death, polyploidy and senescence induced in breast tumor cells by the substituted pyrrole JG-03-14, a novel microtubule poison
Autophagic cell death, polyploidy and senescence induced in breast tumor cells by the substituted pyrrole JG-03-14, a novel microtubule poison
复制标题
DOI：
10.1016/j.bcp.2007.07.003
复制
发表时间：
2007-10-01
影响因子：
5.8
通讯作者：
Gewirtz, David A.
中科院分区：
文献类型：
--
作者：
Arthur, Christopher R.;Gupton, John T.;Gewirtz, David A.
针对MCF-7（p53野生型）、MDA-MB 231（p53突变体）、MCF-7/半胱天冬酶3和MCF-7/ADR（多药耐药）乳腺肿瘤细胞系筛选JG-03-14（抑制微管聚合的取代吡咯）。通过结晶紫染料试验评估细胞活力和生长抑制。凋亡通过TUNEL检测，细胞周期分布通过流式细胞术，自噬通过泡形成的吖啶橙子染色，衰老基于β-半乳糖苷酶染色和细胞形态学。我们的研究表明，暴露于浓度为500 nM的JG-03-14可诱导MCF-7和MDA-MB 231细胞系中的时间依赖性细胞死亡。在MCF-7细胞中，发现残留的存活细胞群体是衰老的;相反，在处理的MDA-MB 231细胞中没有存活的衰老群体。在处理后15天的时间内，在任一细胞系中均未检测到增殖恢复。TUNEL测定和流式细胞术均表明MCF-7细胞中响应于药物处理的相对有限程度的凋亡（< 10%），而MDA-MB 231细胞中的凋亡更广泛（但< 20%）;指示自噬细胞死亡的酸性空泡形成在MCF-7和MDA-MB 231细胞中均相对广泛。此外，JG-03-14诱导MDA-MB 231细胞中形成大的超二倍体细胞群。JG-03-14在MCF-7/caspase 3细胞和MCF-7/ADR细胞系中也表现出显著的抗增殖活性。JG-03-14促进自噬性细胞死亡并且还在表达多药耐药泵的肿瘤细胞中保持活性的观察结果表明，取代的吡咯类的新型微管毒物可能有望用于治疗乳腺癌。(c)2007年爱思唯尔公司All rights reserved.
JG-03-14, a substituted pyrrole that inhibits microtubule polymerization, was screened against MCF-7 (p53 wild type), MDA-MB231 (p53 mutant), MCF-7/caspase 3 and MCF-7/ADR (multidrug resistant) breast tumor cell lines. Cell viability and growth inhibition were assessed by the crystal violet dye assay. Apoptosis was evaluated by the TUNEL assay, cell cycle distribution by flow cytometry, autophagy by acridine orange staining of vesicle formation, and senescence based on p-galactosidase staining and cell morphology. Our studies indicate that exposure to JG-03-14, at a concentration of 500 nM, induces time-dependent cell death in the MCF-7 and MDA-MB231 cell lines. In MCF-7 cells, a residual surviving cell population was found to be senescent; in contrast, there was no surviving senescent population in treated MDA-MB231 cells. No proliferative recovery was detected over a period of 15 days post-treatment in either cell line. Both the TUNEL assay and FLOW cytometry indicated a relatively limited degree of apoptosis (< 10%) in response to drug treatment in MCF-7 cells with more extensive apoptosis (but < 20%) in MDA-MB231 cells; acidic vacuole formation indicative of autophagic cell death was relatively extensive in both MCF-7 and MDA-MB231 cells. In addition, JG-03-14 induced the formation of a large hyperdiploid cell population in MDA-MB231 cells. JG-03-14 also demonstrated pronounced anti-proliferative activity in MCF-7/caspase 3 cells and in the MCF-7/ADR cell line. The observation that JG-03-14 promotes autophagic cell death and also retains activity in tumor cells expressing the multidrug resistance pump indicates that novel microtubule poisons of the substituted pyrroles class may hold promise in the treatment of breast cancer. (c) 2007 Elsevier Inc. All rights reserved.