【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。更多还原

【Abstract】 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.更多还原