三氧化二砷抑制小鼠肝癌 H22 细胞在体、外转移潜能
Arsenic trioxide inhibits metastatic potential of mouse hepatoma H22 cells in vitro and in vivo.
机构信息
Department of Gastroenterology, Fourth Affiliated Hospital, Harbin Medical University, Harbin 150010, China.
出版信息
Hepatobiliary Pancreat Dis Int. 2009 Oct;8(5):510-7.
BACKGROUND
It has been pointed out that only low-dose arsenic trioxide (ATO) presents therapeutic benefits outweighing the toxic side effects. Low-dose ATO can effectively alleviate acute promyelocytic leukemia (APL). However, it is quite challenging in treating solid tumors. The purpose of this study was to investigate the effect of ATO at low concentrations on the metastatic potential of mouse hepatoma H(22) cells and the anti-metastatic mechanism of ATO.
METHODS
The metastatic potential of H(22) cells was evaluated by adhesion, migration and invasion assays after exposure to a low dose of ATO in vitro. The mouse lung metastatic model induced by injection of H(22) cells via the tail vein was adopted for the evaluation of metastatic potential. Different proteins in the lysate of H(22) cells exposed to ATO at different concentrations were investigated by surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Finally, Western blotting analyses were made to detect the expression pattern of MMP-2 and nm23-M1 proteins.
RESULTS
Significant cell death started at ATO concentrations above 2 micromol/L. The growth and adhesion potential of H(22) cells was inhibited in a time- and dose-dependent manner, and the migration and invasion potential of H(22) cells was inhibited in a dose-dependent manner while ATO concentration was below 2 micromol/L. Mice injected with ATO at a dose of 0.5 mg/kg had fewer lung metastases. However, mice injected with ATO at a dose of 2 mg/kg or 4 mg/kg had a high mortality rate and more liver injuries. A total of 15 different protein peaks were identified between the lysate of H(22) cells treated with ATO and controls. Two proteins that peaked at m/z 5302 and 17207 coincided with MMP-2 (fragment) and nm23-M1, respectively. Western blotting analyses demonstrated that MMP-2 and MMP-2 fragments were down-regulated and nm23-M1 was up-regulated in H(22) cells treated with 2 micromol/L ATO for 48 hours.
CONCLUSIONS
ATO at a low dose inhibits the metastatic potential of mouse hepatoma H(22) cells in vitro and in vivo, and involves down-regulation of MMP-2 and up-regulation of nm23-M1.
背景
有研究指出，只有低剂量的三氧化二砷（ATO）才具有治疗效益超过毒性副作用的优势。低剂量的 ATO 能有效缓解急性早幼粒细胞白血病（APL），但在治疗实体瘤方面却极具挑战性。本研究旨在探讨低浓度 ATO 对小鼠肝癌 H（22）细胞转移潜能的影响及其抗转移机制。
方法
通过体外接触低剂量 ATO 后进行黏附、迁移和侵袭实验，评估 H（22）细胞的转移潜能。通过尾静脉注射 H（22）细胞诱导小鼠肺转移模型，评估转移潜能。采用表面增强激光解吸电离飞行时间质谱（SELDI-TOF-MS）检测不同浓度 ATO 作用于 H（22）细胞后细胞裂解物中的不同蛋白。最后，采用 Western 印迹分析检测 MMP-2 和 nm23-M1 蛋白的表达模式。
结果
ATO 浓度超过 2 微米时，细胞开始出现明显的死亡。H（22）细胞的生长和黏附能力呈时间和剂量依赖性抑制，而在 ATO 浓度低于 2 微米时，H（22）细胞的迁移和侵袭能力呈剂量依赖性抑制。以 0.5mg/kg 剂量注射 ATO 的小鼠肺部转移灶较少，但以 2mg/kg 或 4mg/kg 剂量注射 ATO 的小鼠死亡率较高，肝损伤较多。在 ATO 处理的 H（22）细胞裂解物与对照组之间共鉴定出 15 个不同的蛋白峰。两个蛋白峰在 m/z 5302 和 17207 处的分子量分别与 MMP-2（片段）和 nm23-M1 相对应。Western 印迹分析表明，2 微米 ATO 处理 48 小时后，H（22）细胞中的 MMP-2 和 MMP-2 片段下调，nm23-M1 上调。
结论
低剂量 ATO 可抑制体内外小鼠肝癌 H（22）细胞的转移潜能，其机制涉及 MMP-2 的下调和 nm23-M1 的上调。
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