Doxycycline抑制人肝癌细胞系HepG_2生长的实验研究  

Doxycycline inhibits the growth of human hepatoma cell HepG_2 in vitro

作　　者：张强波[1] 李杰[1] 时昌文[1] 李捷[1] 孙京杰[1] 曹莉莉[1] 

机构地区：[1]山东大学附属千佛山医院普外中心,山东济南250014

出　　处：《中国现代普通外科进展》2008年第2期156-159,110,共5页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:观察多西环素(Doxycycline)对人肝细胞性肝癌细胞系HepG2生长抑制和凋亡的作用,并初步探讨其作用机制。方法:不同浓度的Doxycycline(5、10、20、30和50mg/L)对体外培养人肝癌细胞系HepG2进行不同的时间干预(24、48和72h),MTT法测定细胞生长抑制率,TUNEL法检测细胞凋亡率,免疫细胞化学检测Fas、FasL及MMP-2蛋白表达的变化。结果:Doxycycline作用HepG2细胞48h和72h后,细胞生长被明显抑制,与无药对照组比较差异有统计学意义(P<0.001),且呈时间、浓度依赖趋势。其细胞凋亡率较无药对照组也明显升高(P<0.01);20mg/L Doxycycline作用于癌细胞48h后,实验组FasL蛋白阳性表达较无药对照组明显升高,MMP-2蛋白阳性表达较无药对照组明显降低,两组差异均有统计学意义(P<0.01),而Fas蛋白表达则差异无统计学意义(P>0.05)。结论:Doxycycline对人肝癌细胞系HepG2具有明显的生长抑制和促凋亡作用,其机制可能与下调MMP-2、上调FasL从而启动Fas/FasL膜受体途径有关。Objective： To observe the effect and mechanism of doxycycline on cell growth inhibition and the apoptosis of human hepatoma cell HepG2 in vitro. Methods＇. HepG2 cells were treated with doxycycline at different concentrations and time in vitro. Cell growth inhibition rate was detected by MTT assay, and apoptosis rate was identified by TUNEL assay, the expression of Fas,FasL and MMP-2 proteins were examined by immunocytochemical staining, Results： Treated with doxycycline at different concentrations （5,10,20,30 and 50 mg·L^-1） for 48 and 72 hours, the growth of HepG2 cells decreased significantly （P〈0.001） and there was a tendency of dose-time dependent manner. Incubated HepG2 cells with doxycycline at 20 rag. L-1 for 48 hours, the expression of FasL were up-regulated and MMP-2 were down-regulated compared with those of control group （P〈0. 01）, while the difference of Fas expression was insignificant （P〉0.05）. Conclusion： Doxycycline might inhibit cell growth and induce apoptosis of HepG2 significantly. The mechanism of doxycycline induced apoptosis might be associated with down-regulating expression of MMP-2 and up-regu- lating expression of FasL on HepG2 cells.

关 键 词：肝肿瘤 DOXYCYCLINE 细胞凋亡 FASL MMP-2 

分 类 号：R735.7[医药卫生—肿瘤]