苦丁茶熊果酸诱导鼻咽癌细胞凋亡及其对Survivin和 APC蛋白表达的影响

摘要

目的 探讨不同浓度的苦丁茶熊果酸诱导鼻咽癌细胞凋亡情况及其对大肠腺瘤样息肉(adenomatous polyosis coli,APC)和生存素(Survivin)蛋白表达的影响,为揭示苦丁茶熊果酸的抗肿瘤机制提供研究依据.方法 从苦丁茶老叶中提取熊果酸并纯化与鉴定,以10、20、40、80、100 μmol/L的苦丁茶熊果酸浓度持续作用于在培养的鼻咽癌细胞24 h后,流式细胞术分析细胞凋亡率,免疫印迹法(Western blot)检验Survivin和APC蛋白的表达情况.结果 经过不同梯度浓度的苦丁茶熊果酸24 h的持续作用,鼻咽癌细胞发生了不同程度的凋亡,凋亡率呈浓度依赖性,浓度达40 μmol/L时作用效果逐渐明显;Western blot结果显示,随着苦丁茶熊果酸作用时间的延长,Survivin从有表达向表达消失的趋势转变,而APC蛋白的表达则与Survivin相反.结论 苦丁茶熊果酸能诱导鼻咽癌细胞发生凋亡,其机制可能与上调APC蛋白的表达量有关.%Objective To investigate the effects of Kuding tea ursolic acid on apoptosis of human nasopharyngeal carcinoma ( NPC ) cells and expression of survivin and ( adenomatous polyosis coli, APC ) protein, providing evidence for pathogenic study of preventive effect of Kuding tea on NPC. Methods Human NPC cells were cultured with ursolic acid at a concentrations of 10, 20, 40 , 80 and 100 μmol/L for 24 h. Flow cytometry was applied to assess apoptosis induced by ursolic acid, while Western blot was used to measure the expression of survivin and APC protein. Results Significant apoptosis was observed in cells treated with ursolic acid. Increase of APC and reduction of surviving were revealed after treatment. Conclusion The ursolic acid from Kuding tea inhibits proliferation of human NPC cell, possibly via up - regulation of APC.