香加皮杠柳苷诱导人食管癌细胞凋亡及其作用机制的研究  

Periplocin (a sort of ethanol from Cortex periplocae) induces apoptosis of esophageal carcinoma cells by influencing expression of related genes

作　　者：赵连梅[1] 艾军[1] 张倩[1] 韩丽娜[1] 刘丽华[1] 王玲[1] 单保恩[1] 

机构地区：[1]河北医科大学第四医院科研中心,石家庄050011

出　　处：《肿瘤》2009年第11期1025-1030,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:30772752);河北省自然科学基金资助项目(编号:C2004000610)

摘　　要：目的:探讨中药香加皮醇提物杠柳苷诱导人食管癌细胞凋亡及其作用机制。方法:应用MTT法检测香加皮醇提物杠柳苷对食管癌细胞TE-13、Eca-109、TE-1和TE-10增殖的影响,并计算其对食管癌细胞的半数抑制浓度(half inhibition concentration,IC50);在光学显微镜下观察经杠柳苷处理后细胞的形态学改变;Wright-Giemsa染色观察Eca-109细胞凋亡的形态学变化;FCM检测杠柳苷对Eca-109细胞凋亡的影响;RT-PCR法检测经杠柳苷处理后细胞凋亡相关基因mRNA表达的改变;caspase-3/7蛋白酶活性检测试剂盒检测细胞caspase蛋白酶活性的变化。结果:香加皮醇提物杠柳苷对人食管癌细胞的增殖具有明显的抑制作用;经杠柳苷处理后,Eca-109细胞出现明显的凋亡形态学变化,与未处理组比较,细胞凋亡率明显上升(P<0.05);经杠柳苷处理后,Eca-109细胞中survivin和bcl-2 mRNA的表达下调(P<0.05),bax mRNA的表达水平升高(P<0.05),xiap、caspase-3和caspase-7 mRNA的表达水平无明显变化,而caspase-3/7蛋白酶活性明显增加(P<0.05)。结论:杠柳苷可明显抑制食管癌细胞的增殖,其作用机制可能是通过下调凋亡抑制基因的表达而诱导细胞凋亡。Objective ： To study the effect of periplocin （ a sort of ethanol from Cortex periplocae） on apoptosis of human esopha- geal carcinoma cell lines and elucidate its action mechanism. Methods： The proliferation of esophageal cancer cell lines TE-13, Eca- 109, TE-1, and TE-10 were measured using MTF method. The half inhibition concentration （1C50） of periplocin was calculated. Mor- phological changes of esophageal carcinoma cells were observed under light microscope before and after periplocin treatment. The mor- phological features of apoptosis of Eca-109 cells were observed by Giemsa staining. FCM was used to detect apoptotic rate of Eca-109 cells induced by periplocin. Effect of periplocin on mRNA transcription of apoptosis-related genes was determined by RT-PCR assays. Caspase 3/7 activity was detected by caspase Glo 3/7 kit. Results： Periplocin markedly inhibited the proliferation of esophageal cancer cells . After treatment with periplocin, Eca-109 showed apparent apoptotic morphological changes and the apoptosis rate was increased （P 〈 0.05 ）. Expression levels of survivin and bcl-2 mRNA in Eca-109 were down-regulated, but bax mRNA expression was up-regula- ted after periplociu treatment （both P 〈 0.05）. The expression levels of xiap,caspase 3, and caspase 7 mRNA were not changed, but the activity of caspase 3/7 was enhanced significantly as compared to control group （P 〈 0.05 ）. Conclusion ： Periplocin showed very strong inhibitory effects on the proliferation of esophageal carcinoma cells, and the mechanism may be related with down-re-gulation of the expression of apoptosis-suppressor genes and induction of apoptosis.

关 键 词：食管肿瘤 细胞凋亡 香加皮杠柳苷 细胞 ECA-109 

分 类 号：R733.72[医药卫生—肿瘤] R73-352[医药卫生—临床医学]