丹皮酚对人肝癌BEL-7402细胞凋亡和COX-2、Survivin、XIAP、c-IAP1表达的影响  

Effects of paenonal on apoptosis of hepatocellular carcinoma BEL-7402 cell lines and expressions of COX-2,Survivin,XIAP and c-IAP1

作　　者：彭万仁[1] 付卫争[1] 孙国平[1] 范璐璐[1] 葛磊[1] 吴志丽[1] 

机构地区：[1]安徽医科大学第一附属医院肿瘤内科,安徽合肥230022

出　　处：《中国药理学通报》2010年第6期735-739,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30772537)

摘　　要：目的研究丹皮酚(paenonal,Pae)对COX-2及凋亡抑制蛋白Survivin、XIAP、c-IAP1表达的影响,探讨其诱导人肝癌BEL-7402细胞凋亡的作用机制。方法应用MTT法检测不同浓度的Pae对体外培养的人肝癌BEL-7402细胞增殖抑制作用,流式细胞术和原位细胞凋亡检测法(TUNEL)检测细胞凋亡,免疫细胞化学技术检测COX-2、Survivin、XIAP、c-IAP1的表达情况。结果 Pae可以明显抑制BEL-7402细胞的增殖,并随着剂量的增加和时间的延长,该抑制作用增强,经7.81、15.63、31.25、62.5mg·L-1Pae作用72h后,其抑制率分别为20.54%、25.74%、33.04%、57.97%(P<0.01或P<0.05);TUNEL染色可见典型的凋亡形态学特征包括染色质凝集、边集化,呈新月形聚集于核膜下;流式细胞仪检测用药组凋亡率明显高于对照组,Pae浓度为7.81、15.63、31.25、62.5mg·L-1时,其凋亡率分别为(1.75±0.32)%、(8.26±0.95)%、(12.46±1.37)%、(25.07±1.87)%、(37.52±2.56)%。与对照组相比,免疫细胞化学分析显示,COX-2、Survivin、XIAP、c-IAP1表达明显下调(P<0.01)。结论 Pae对人肝癌细胞株BEL-7402有抑制增殖和诱导凋亡的作用,其机制可能与通过下调COX-2的表达并与抑制survivin、XIAP和c-IAP1的表达有关。Aim To investigate the effect of paeonol on Expressions of COX-2,surviving,XIAP,c-IAP1 and its mechanism of inducing hepatocarcinoma cells apoptosis.Methods The inhibitory effect of Paeonol with different concentration on hepatocarcinoma cells in vitro was detected by MTT assay.The apoptosis was detected by flow cytometry（FCM）and TUNEL.Expressions of COX-2,survivin,XIAP and c-IAP1 were analyzed by immunocytochemical staining.Results Pae significantly inhibited cell proliferation in time-and dose-dependent manner.When interfered with Pae at a dose of 7.81,15.63,31.25,62.5 mg·L-1 for 48 h,the inhibiting rates were 20.54%,25.74%,33.04% and 57.97%（P0.01或P0.05）,respectively.The cells showed distinctive apoptotic characteristics included chromatin condensation and deformed and fragmented nuclei by TUNEL.The apoptosis rate of the group treated with Pae was higher than those of the control group.When Pae was at different concentrations（7.81,15.63,31.25,62.5 mg·L-1）,the results were （1.75±0.32）%,（8.26±0.95）%,（12.46±1.37）%,（25.07±1.87）%,（37.52±2.56）%,respectively.Compared with those of control group,the expressions of COX-2,survivin,XIAP and c-IAP1 were significantly down-regulated in BEL-7402 cells by Pae.Conclusion Pae can inhibit the proliferation and induce apoptosis of BEL-7402 Cell Lines,which may be fulfilled by down-regulation of the COX-2,survivin,XIAP and c-IAP1.

关 键 词：丹皮酚 肝细胞癌 环氧合酶-2 细胞凋亡 凋亡抑制蛋白 流式细胞术 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]