脂肪酸酰胺水解酶抑制剂URB597对人肝癌细胞MHCC97H生长和侵袭的抑制作用及机制研究

目的：观察脂肪酸酰胺水解酶抑制剂URB597对人肝癌高转移细胞MHCC97H细胞生长和侵袭性的抑制作用。方法：不同浓度URB597（1、5、10μmol/L）作用于MHCC97H细胞后不同时间,应用MTT法及流式细胞仪检测该种细胞生长活力及凋亡细胞数目的改变;应用Transwell实验检测该种细胞的运动能力及侵袭能力;应用蛋白质印迹法检测该种细胞内磷酸化Akt（p-Akt）和Akt表达量的变化。结果：上述3种浓度的URB597作用于该种细胞3-7d后,细胞活力明显降低,呈时间及剂量依赖性;10μmol/L URB597作用3-7d可显著增加凋亡细胞数目。URB597作用于细胞48h后,与对照组比较,5、10μmol/L URB597可显著抑制MHCC97H细胞侵袭能力（P〈0.01,P〈0.001）。3种浓度URB597作用于细胞24h后,5、10μmol/L URB597可显著下调细胞内p-Akt水平（P〈0.05,P〈0.01）。结论：URB597对体外生长的人肝癌细胞MHCC97H的生长和侵袭有明显抑制作用,该作用可能与其抑制磷酯酰肌醇-3激酶（PI3K）/Akt信号通路有关。

Inhibitory effects of fatty acid amide hydrolase inhibitor URB597 on the proliferation and invasion of human hepatoma cell MHCC97H and its mechanism

Objective： To evaluate the inhibitory effects of fatty acid amide hydrolase inhibitor URB597 on the proliferation and invasion of human hepatoma cell line MHCC97H. Methods： MTT method and flow cytometry were used to analyze the effects of different concentrations of URB597（1,5,10 μmol/L） on the changes of cell active viability and apoptosis cell number of MHCC97H at different time points. Transwell test was used to investigate the movement and invasion of the cells. The lev- els of phosphated Akt（p-Akt） and Akt with or without treatment by URB597 were detected by Western-blot assay. Results： Three to seven days after treatment of the above concentrations of URB597 in this type of cell, cell growth was significantly inhibited, with time and concentration dependence, and 3-7 day 10μmol/L URB597 treatment could significantly increase the number of apoptotic cells. Forty-eight hours after treatment with URB597,5,10 μmol/L URB597 could obviously suppress the invasion of MHCC97H cells （P〈0. 01, P〈0. 001）, when compared with those of the control group. Twenty-four hours after treatment of 3 different concentrations of URB597,5,10 μmol/L URB597 could obviously down-regulate the concentration of p Akt （P〈0. 01, P〈0. 001）. Conclusion： URB597 seemed to have obvious inhibitory effects on the proliferation and invasion of MHCC97H cells in vitro, which might be associated with the inhibition of phosphatidylinositol 3-kinase （PI3K）/Akt signal pathway.