亲脂性芳酰腙螯合剂 HNTMB 及其对卵巢癌细胞的多种作用。

Lipophilic aroylhydrazone chelator HNTMB and its multiple effects on ovarian cancer cells.

机构信息

Molecular Therapeutics Laboratory, Program in Women's Oncology, Department of Obstetrics and Gynecology, Women and Infants' Hospital of RI, Alpert Medical School of Brown University, Providence, RI 02905, USA.

出版信息

BMC Cancer. 2010 Feb 25;10:72. doi: 10.1186/1471-2407-10-72.

DOI:10.1186/1471-2407-10-72

PMID:20184758

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2836302/

Abstract

BACKGROUND

Metal chelators have gained much attention as potential anti-cancer agents. However, the effects of chelators are often linked solely to their capacity to bind iron while the potential complexation of other trace metals has not been fully investigated. In present study, we evaluated the effects of various lipophilic aroylhydrazone chelators (AHC), including novel compound HNTMB, on various ovarian cancer cell lines (SKOV-3, OVCAR-3, NUTU-19).

METHODS

Cell viability was analyzed via MTS cytotoxicity assays and NCI60 cancer cell growth screens. Apoptotic events were monitored via Western Blot analysis, fluorescence microscopy and TUNEL assay. FACS analysis was carried out to study Cell Cycle regulation and detection of intracellular Reactive Oxygen Species (ROS) RESULTS: HNTMB displayed high cytotoxicity (IC50 200-400 nM) compared to previously developed AHC (oVtBBH, HNtBBH, StBBH/206, HNTh2H/315, HNI/311; IC50 0.8-6 microM) or cancer drug Deferoxamine, a hexadentate iron-chelator (IC50 12-25 microM). In a NCI60 cancer cell line screen HNTMB exhibited growth inhibitory effects with remarkable differences in specificity depending on the cell line studied (GI50 10 nM-2.4 microM). In SKOV-3 ovarian cancer cells HNTMB treatment led to chromatin fragmentation and activation of the extrinsic and intrinsic pathways of apoptosis with specific down-regulation of Bcl-2. HNTMB caused delayed cell cycle progression of SKOV-3 through G2/M phase arrest. HNTMB can chelate iron and copper of different oxidation states. Complexation with copper lead to high cytotoxicity via generation of reactive oxygen species (ROS) while treatment with iron complexes of the drug caused neither cytotoxicity nor increased ROS levels.

CONCLUSIONS

The present report suggests that both, non-complexed HNTMB as a chelator of intracellular trace-metals as well as a cytotoxic HNTMB/copper complex may be developed as potential therapeutic drugs in the treatment of ovarian and other solid tumors.

摘要

背景

金属螯合剂作为潜在的抗癌药物受到了广泛关注。然而，螯合剂的作用通常仅与其结合铁的能力相关，而其他痕量金属的潜在络合作用尚未得到充分研究。在本研究中，我们评估了各种亲脂性芳酰腙螯合剂（AHC），包括新型化合物 HNTMB，对各种卵巢癌细胞系（SKOV-3、OVCAR-3、NUTU-19）的影响。

方法

通过 MTS 细胞毒性测定和 NCI60 癌症细胞生长筛选分析细胞活力。通过 Western Blot 分析、荧光显微镜和 TUNEL 测定监测凋亡事件。通过 FACS 分析研究细胞周期调控和检测细胞内活性氧物种（ROS）。

结果

与先前开发的 AHC（oVtBBH、HNtBBH、StBBH/206、HNTh2H/315、HNI/311；IC50 0.8-6 microM）或癌症药物去铁胺（一种六齿铁螯合剂，IC50 为 12-25 microM）相比，HNTMB 显示出高细胞毒性（IC50 为 200-400 nM）。在 NCI60 癌症细胞系筛选中，HNTMB 表现出生长抑制作用，其特异性因所研究的细胞系而异（GI50 为 10 nM-2.4 microM）。在 SKOV-3 卵巢癌细胞中，HNTMB 处理导致染色质片段化和细胞凋亡的外在和内在途径的激活，同时特异性地下调 Bcl-2。HNTMB 通过 G2/M 期阻滞导致 SKOV-3 细胞周期延迟。HNTMB 可以螯合不同氧化态的铁和铜。与铜的络合导致活性氧物种（ROS）的产生，从而导致高细胞毒性，而药物的铁络合物处理既不会导致细胞毒性也不会增加 ROS 水平。