靶向PCNA基因的小干扰RNA对鼻咽癌CNE2细胞周期的影响  

Effects of siRNA targeting PCNA gene on nasopharyngeal carcinoma CNE2 cell cycle

作　　者：练兵[1] 王继群[2] 金琳[3] 

机构地区：[1]暨南大学附属第一医院耳鼻咽喉科,广东广州510632 [2]暨南大学医学院,广东广州510632 [3]暨南大学生物医药基地,广东广州510632

出　　处：《中国病理生理杂志》2009年第8期1533-1537,共5页Chinese Journal of Pathophysiology

摘　　要：目的:研究小干扰RNA抑制PCNA基因表达后对鼻咽癌CNE2细胞株增殖及细胞周期的影响。方法:利用体外合成法合成针对PCNA基因的小干扰RNA(siRNA)序列,应用脂质体转染鼻咽癌CNE-2细胞,实时定量PCR(real-timePCR)和免疫组织化学法观察siRNA干扰后细胞中PCNA mRNA水平和PCNA蛋白表达,倒置相差显微镜观察转染前后CNE2细胞的生长变化,流式细胞仪检测siRNA干扰后的细胞周期。结果:在siR-NA转染CNE2细胞后,细胞增殖受到抑制,蛋白表达不同程度降低,对PCNA mRNA水平的抑制达98.5%,细胞周期在G0/G1停滞。结论:siRNA可在鼻咽癌中有效发挥RNA干扰效应,抑制PCNA mRNA及蛋白表达,降低PCNA活性,抑制鼻咽癌细胞增殖,为鼻咽癌基因治疗提供了新的方法。AIM： To explore the effects of small interfering RNA （siRNA） targeting PCNA gene on nasopharyngeal carcinoma CNE2 ceils growth and cycle. METHODS ： Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent. The PCNA mRNA and PCNA protein were detected by real - time polymerase chain reaction （RT- PCR） and immunohistochemical method. Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA - siRNA transfected. Flow cytometry was used to observe the cell cycle. RESULTS ： In CNE2 cells after PCNA - siRNA transfection, the expressions of PCNA mRNA and protein were down -regulated at different degree. Inhibition ratio of PCNA mRNA was 98.5%. Meanwhile, the cell cycle was suffocated at G0/G1 stage. CONCLUSION： The synthesized PCNA - siRNA effectively interferes nasopharyngeal carcinoma cells by down - regulating the expressions of the PCNA mRNA and its protein, therefore inhibits the growth of CNE2 cells. Future application of PCNA - siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.

关 键 词：siRNA 鼻咽肿瘤 增殖细胞核抗原 CNE2细胞 

分 类 号：R766[医药卫生—耳鼻咽喉科]