Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status
Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status
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DOI：
10.1023/a:1013855615712
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发表时间：
2002-01-01
影响因子：
3.8
通讯作者：
Woynarowski, JM
中科院分区：
文献类型：
--
作者：
Herzig, MCS;Liang, HY;Woynarowski, JM
Caspase-3 缺乏会限制促凋亡抗癌治疗的效率。 Irofulven（羟甲基酰基富烯，HMAF，MGI 114，NSC 683863）是一种抗肿瘤药物，目前处于 III 期和多个 II 期试验中，可以在凋亡诱导中区分肿瘤和正常细胞。本研究调查了irofulven诱导的细胞凋亡是否需要caspase-3。在不同 caspase-3 状态的乳腺癌细胞（缺陷型 MCF-7 细胞和丰富型 MDA-MB-231 细胞）以及正常人乳腺上皮细胞 HMEC 中比较了 Irofulven 的作用。无论 caspase-3 状态如何，Irofulven 都会在乳腺癌细胞系中诱导显着的、浓度和时间依赖性的细胞凋亡 DNA 断裂。在 1 muM irofulven（类似于 3 x GI(5)0）孵育 12、24 和 48 小时后，MCF-7 和 MDA-MB-231 细胞中片段化 DNA 分别占总 DNA 的 3.7%、14.1% 和 34.6%，以及 8.4%、12.6% 和 20.3%。细胞活力（台盼蓝排除）在前 24 小时内基本不受影响，但在 48 小时后显着下降，表明继发性坏死。 48 小时时细胞数量的净损失很明显。正常 HMEC 细胞对 1 muM 药物无效，12-48 小时后仅具有 3-9% 的 DNA 片段，但在药物水平 > 3 muM 时观察到细胞凋亡。广谱半胱天冬酶抑制剂 Z-VAD-fmk 在 20 muM 浓度下抑制艾洛芬诱导的所有细胞系凋亡，在 100 muM 浓度下几乎完全消除细胞凋亡。 Irofulven 处理导致 MDA-MB-231 和 HMEC 细胞中的边缘 caspase-3 加工。这些结果表明，虽然 caspase 级联介导 irofulven 诱导的细胞凋亡，但 caspase-3 是可有可无的（由 NIH CA70091 和 CA78706 支持）。
Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 muM irofulven (similar to 3 x GI(5)0), fragmented DNA comprised 3.7, 14.1 and 34.6% and 8.4, 12.6 and 20.3% of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 muM drug with only similar to3-9% fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 muM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 muM with nearly complete abrogation of apoptosis at 100 muM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706).