粉背南蛇藤中新三萜化合物对RKO细胞体外抗癌作用的研究  

Antitumor effects of novel triterpene from Celastrus hypoleucus on human colorectal cancer cell line RKO in vitro

作　　者：毛建山[1] 王奎武[2] 郑树[3] 潘远江[2] 

机构地区：[1]浙江大学医学院附属第二医院消化内科,浙江杭州310009 [2]浙江大学化学系,浙江杭州310027 [3]浙江大学肿瘤研究所,浙江杭州310009

出　　处：《中国中药杂志》2006年第17期1450-1453,共4页China Journal of Chinese Materia Medica

基　　金：浙江省中医药科研基金项目(2004C094)

摘　　要：目的：探讨粉背南蛇藤中新的三萜齐墩果-12-烯-3β，6α-二醇对人结肠癌细胞系RKO细胞的体外增殖抑制作用和诱导凋亡作用。方法：采用MTT法检测齐墩果-12-烯-3β，6α-二醇对体外培养人结肠癌细胞系RKO细胞的增殖抑制作用，通过AO／EB双染色荧光显微镜、HE染色光学显微镜、DNA琼脂糖凝胶电泳、流式细胞仪分析齐墩果-12-烯-3β，6α-二醇对RKO细胞的诱导凋亡作用及对细胞周期的影响。结果：齐墩果-12-烯-3β，6α-二醇对RKO细胞生长抑制作用具有剂量依赖性和时间依赖性，作用48h时IC50为（12．20±0．79）μg·mL^-1；AO／EB双染色、HE染色可见典型的肿瘤细胞凋亡改变；10μg·mL^-1的三萜处理RKO细胞48h即可出现明显的DNA梯带、流式细胞仪可检测到亚二倍体峰，在10-20μg·mL^-1内具有明显的量-效作用关系；并可见RKO细胞G0-G1期、G2-M期的细胞明显减少，而S期细胞无变化。结论：粉背南蛇藤中新的三萜齐墩果-12-烯-3β，6α-二醇能抑制人结肠癌细胞系RKO细胞增殖和诱导凋亡。Objective： To investigate the effects of novel triterpene（ 12-oleanene-3β, 6α-diol） from Celastrus hypoleucus on the proliferation and apoptosis of human colorectal cancer cell line RKO. Method： The inhibitory effect of the novel triterpene on RKO cell proliferation was assayed by MTT dye reduction. The morphology of apoptotic cells was observed with AO/EB double fluorescence stai-ning and HE staining, DNA fragment with electrophoresis on agarose gels, sub-diploid peak and cell cycle with flow cytometer（FCM）. Result： Novel triterpene（ 12-oleanene-3β, 6α-diol） from C. hypoleucus significantly inhibited proliferation of RKO cells in dose-dependent and time-dependent manner, the IC50was（ 12.20 ± 0.79） μg·mL^-1at 48 h. Typical apoptotic changes were observed in RKO cells under the fluorescence microscope and the light microscope. DNA ladder was detected on agarose gels at concentrations from 10 μg·mL^-1 to 20 μg·mL^-1 at 48 h. With FCM methods, dose-dependent apoptosis-induced effect was observed in RKO cell line after treatment of triterpene for 48 h, and the apoptotic rates were increased from（2.93 ± 0.84 ） % to（50.79 ± 6.61 ） % at concentrations from 2.5 μg·mL^-1 to 20 μg·mL^-1 DNA histograms data from FCM analysis showed that the number of cells was obviously reduced during G0-G1 phase and G2-M phase, but not during S phase for RKO cell line after treatment with various concentrations of the triterpene for 48 hours. Conclusion： Novel triterpene（ 12-oleanene-3β, 6α-diol） from C. hypoleucus can induce apoptosis and has inhibition effect on the proliferation in RKO cell line.

关 键 词：齐墩果-12-烯-3β 6α-二醇 粉背南蛇藤 抗肿瘤 细胞凋亡 RKO细胞 

分 类 号：R285.5[医药卫生—中药学]