β-lapachone induces cell cycle arrest and apoptosis in human colon cancer cells
β-lapachone induces cell cycle arrest and apoptosis in human colon cancer cells
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DOI：
10.1007/bf03402095
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发表时间：
1999-11-01
影响因子：
5.7
通讯作者：
Pardee, AB
中科院分区：
文献类型：
--
作者：
Huang, LL;Pardee, AB
背景资料：人类结肠癌具有高频率的p53突变，并且表达突变型p53的癌细胞倾向于对当前的化疗和放疗具有抗性。因此，重要的是找到可以抑制具有改变的p53状态的结肠癌细胞的治疗剂。β-拉帕酮是一种新型拓扑异构酶抑制剂，已显示通过p53非依赖性途径诱导人早幼粒细胞白血病和前列腺癌细胞的细胞死亡。材料和方法：采用p53突变或缺陷的人结肠癌细胞系SW 480、SW 620和DLD 1，研究β-拉帕醌对人结肠癌细胞的作用。通过集落形成试验、细胞周期分析和凋亡分析（包括膜联蛋白V染色和DNA梯状分析）来评估β-拉帕酮的抗增殖作用。对细胞周期和凋亡调控蛋白的影响进行了检查，通过immunoblotting.Results：所有三个细胞系，SW 480，SW 620，和DLD 1，是敏感的β-拉帕醌，与IC 50的2至3 μ M的克隆形成试验，发现类似于以前报道的前列腺癌细胞。然而，这些细胞被阻滞在S期的不同阶段。在处理后24小时，β-拉帕酮分别诱导SW 480、SW 620和DLD 1细胞的S期、晚期S/G2期和早期S期停滞。β-拉帕酮诱导的细胞周期改变与细胞周期调节蛋白如细胞周期蛋白A、细胞周期蛋白B1、cdc 2和细胞周期蛋白D1的变化一致。此外，β-拉帕醌诱导细胞凋亡，如膜联蛋白V染色，现在DNA含量的细胞计数分析，和DNA梯状分析所示。此外，β-拉帕醌下调SW 480细胞中突变型p53和诱导p27表达，以及诱导DLD 1细胞中促凋亡蛋白Bar表达，可能与其抑制细胞增殖和诱导细胞凋亡有关。结论：β-拉帕醌通过非p53依赖性途径诱导人结肠癌细胞周期阻滞和凋亡。对于通常含有p53突变的人类结肠癌，β-拉帕酮可能被证明是一种有前途的抗癌剂，可以靶向癌细胞，特别是那些具有突变p53的癌细胞。
Background: Human colon cancers have a high frequency of p53 mutations, and cancer cells expressing mutant p53 tend to be resistant to current chemo- and radiation therapy. It is thus important to find therapeutic agents that can inhibit colon cancer cells with altered p53 status. beta-Lapachone, a novel topoisomerase inhibitor, has been shown to induce cell death in human promyelocytic leukemia and prostate cancer cells through a p53-independent pathway. Here we examined the effects of beta-lapachone on human colon cancer cells.Materials and Methods: Several human colon cancer cell Lines, SW480, SW620, and DLD1, with mutant or defective p53, were used. The antiproliferative effects of beta-lapachone were assessed by colony formation assays, cell cycle analysis, and apoptosis analysis, including annexin V staining and DNA laddering analysis. The effects on cell cycle and apoptosis regulatory proteins were examined by immunoblotting.Results: All three cell lines, SW480, SW620, and DLD1, were sensitive to beta-lapachone, with an IC50 of 2 to 3 mu M in colony formation assays, a finding similar to that previously reported for prostate cancer cells. However, these cells were arrested in different stages of S phase. At 24 hr post-treatment, beta-lapachone induced S-, late S/G2-, and early S-phase arrest in SW480, SW620, and DLD1 cells, respectively. The cell cycle alterations induced by beta-lapachone were congruous with changes in cell cycle regulatory proteins such as cyclin A, cyclin B1, cdc2, and cyclin D1. Moreover, beta-lapachone induced apoptosis, as demonstrated by annexin V staining, now cytometric analysis of DNA content, and DNA laddering analysis. Furthermore, down-regulation of mutant p53 and induction of p27 in SW480 cells, and induction of pro-apoptotic protein Bar in DLD1 cells may be pertinent to the anti-proliferative and apoptotic effects of beta-lapachone on these cells.Conclusions: beta-Lapachone induced cell cycle arrest and apoptosis in human colon cancer cells through a p53-independent pathway. For human colon cancers, which often contain p53 mutations, beta-lapachone may prove to be a promising anticancer agent that can target cancer cells, especially those with mutant p53.