佛波醇脂和溴脱氧尿嘧啶核苷促进鼻咽癌细胞中CD133的表达

TPA and BrdU promote CD133 expression in nasopharyngeal carcinoma cells

目的:了解促癌剂佛波醇酯(12-O-tetradecanoyl-phorbol-13-acetate,TPA)和非放射性标记物溴脱氧尿嘧啶核苷(bromodeoxyuridine,BrdU)对鼻咽癌细胞5-8F中CD133表达的影响。方法:BrdU、TPA单独作用和Brdu+TPA联合作用处理鼻咽癌细胞株5-8F,不同药物处理组细胞采用实时荧光定量PCR(real-time fluorogentic quantitative PCR,RFQ-PCR)及Western印迹法检测CD133基因mRNA和蛋白水平的表达,FCM法分离并观察CD133+细胞含量的变化,Boyden小室侵袭实验检测细胞侵袭能力的变化。结果:与未处理组细胞相比,BrdU单独作用组和BrdU+TPA联合作用组细胞的CD133 mRNA表达量都有不同程度升高(P值分别为0.037和0.003),TPA单独作用组的CD133 mRNA表达量则降低;Western印迹法检测结果提示,3组药物处理组细胞CD133蛋白的表达量均增高;FCM法检测结果显示,CD133+细胞的含量增加;细胞侵袭能力增强,以BrdU+TPA联合作用组效果最显著。结论:在鼻咽癌细胞中,TPA和BrdU均能提高CD133蛋白的表达,并且有相互协同作用。

Objective：To explore whether tumor-inducing agent 12-O-tetradecanoyl-phorbol-13-acetate （TPA） and bromodeoxyuridine （BrdU） affect CD133 expression in nasopharyngeal carcinoma （NPC） 5-8F cells. Methods：NPC cell line 5-8F was treated with single TPA,single BrdU,or TPA plus BrdU,respectively. CD133 mRNA and protein expression levels were detected by real-time fluorogentic quantitative PCR and Western blotting,respectively. Flow cytometry was used to separate CD133-positive cells and determine their levels. Boyden chamber test was used to measure the invasion capability of the cells. Results：Compared with untreated group,CD133 mRNA levels were increased in single BrdU group and BrdU plus TPA group （P=0.037 and 0.003,respectively）,and decreased in single TPA group. Western blotting indicated that the expressions of CD133 protein was increased in all the three treated groups,and FCM showed that the quantity of CD133-positive cells also increased. The invasion capability was enhanced,especially in BrdU plus TPA group. Conclusion：Both TPA and BrdU increased CD133 expression in NPC.The effects of TPA and BrdU are synergestic.