Procaine has anti-proliferative effect and functions as an inhibitor of DNA methylation in human hepatocellular carcinoma and gastric cancer

普鲁卡因在人肝细胞癌和胃癌中具有抗增殖作用和 DNA 甲基化抑制剂的作用

基本信息

批准号：
17590613
负责人：
FUKAI Kenichi
金额：
$ 2.24万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

In this study, we revealed that a local anesthetic, procaine (PCA), possessed growth-inhibitory and demethylating effect on human hepatoma cells (HLE, HuH7, HepG2, Hep3B, PLC/PRF/5, HuH6) and gastric cancer cells (AGS, MKN1, MKN45, MKN74, KATOIII) in vitro. The viability of PCA-treated cells with or without trichostatin A (TSA) was assessed by MTS assay and found that the viability of HLE, HuH7, HuH6 and all gastric cancer cells examined was significantly decreased by PCA treatment. In these cells, the combination treatment with TSA and PCA exhibited stronger reduction of the viability. To clarify the mechanism of the anti-proliferating effect of PCA, TUNEL assay, FACS analysis and morphological observation of PCA-treated cells were performed. Inhibition of S/G2/M transition, morphological changes such as vacuolation and no increase in apoptosis rate were observed in the PCA-treated HLE cells. The genes that were transcriptionally suppressed by DNA hypermethylation in hepatoma cells or gastric cancer cells were demonstrated to be demethylated and reactivated after PCA treatment. The growth-inhibitory effect of PCA in vivo was tested in nude mice bearing xenograft and PCA treatment was revealed to lead to a significant reduction in tumor volume in vivo. In conclusion, these data indicated that PCA had growth-inhibitory and demethylating effects on human hepatoma cells and gastric cancer cells in vitro and in vivo. PCA may be a candidate agent for future therapies against HCC and gastric cancer.

在这项研究中，我们透露，局部麻醉剂（PCA）对人肝癌细胞具有抑制性和脱甲基作用（HLE，HUH7，HEPG2，HEPG2，HEP3B，PLC/PRF/PRF/5，HUH6）和胃癌细胞（AGS，MKN1，MKN1，MKN45，MKN45，MKN74，KATITOI）。通过MTS分析评估了PCA处理的带有或没有毛乳杆菌A（TSA）的PCA处理的细胞的生存能力，发现HLE，HUH7，HUH6的生存能力和所有检查的所有胃癌细胞的生存能力通过PCA处理均显着降低。在这些细胞中，使用TSA和PCA的组合处理表现出更强的生存能力。为了阐明PCA的抗增殖作用的机制，进行了PCA处理的细胞的TUNEL分析，FACS分析和形态观察。在PCA处理的HLE细胞中，抑制S/G2/M转变，形态学变化，例如空泡和凋亡率没有增加。证明，在PCA处理后，被DNA高甲基化在肝癌细胞或胃癌细胞中被DNA高甲基化抑制的基因被证明是被脱甲基化的并重新激活的。在带有异种移植的裸鼠中测试了PCA在体内的生长抑制作用，并且揭示了PCA治疗可显着减少体内肿瘤体积。总之，这些数据表明PCA在体外和体内对人肝癌细胞和胃癌细胞具有抑制性和脱甲基作用。 PCA可能是针对HCC和胃癌的未来疗法的候选药物。