靶向PCNA基因的小干扰RNA对鼻咽癌CNE2细胞周期的影响

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-time PCR) 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siRNA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。

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

AIM: To explore the effects of small interfering RNA (siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells 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．