【摘要】 目的:探讨siRNA干扰Pokemon基因对鼻咽癌CNE-2细胞生长作用的影响。方法:利用siRNA预测软件得到3条siRNA编码DNA序列,并制备合成siRNA片段,利用RT-PCR和蛋白质印迹法检测其沉默Pokemon基因的效果,筛选出沉默效果最佳的siRNA片段。利用MTT法和流式细胞术检测CNE-2细胞生长与凋亡情况。结果:筛选出一条≥50%沉默效应的siRNA片段,转染鼻咽癌CNE-2细胞后,Pokemon基因表达明显下降(P<0.05),p14ARF和p53基因表达上调,P<0.05。鼻咽癌CNE-2细胞增殖受到抑制(P=0.015),凋亡率升高,P=0.008。结论:Pokemon基因沉默后可抑制CNE-2细胞生长,促进细胞凋亡,其机制可能与p14ARF和p53的表达上调有关。更多还原

【Abstract】 OBJECTIVE: To construct recombinant plasmids containing microRNA targeting the Pokemon gene and investigate the effects of miRNA-mediated downregulation of the Pokemon gene on the proliferation of CNE-2 cells.METHODS: Three siRNA were designed using siRNA prediction software.The best one was chosen according to the result of Pokeman gene silencing through RT-PCR and Western blot.The expression of ARF and p53 gene was observed,and then the influence of CNE-2 cell growth and apotosis were detected by MTT and flow cytometry.RESULTS: After silencing Pokeman gene,the expression of Pokeman gene of nasopharyngeal carcinoma CNE-2 cell was obviously downregulated(P<0.05),but the expression of p14ARF and p53 gene was significantly increased(P<0.05).The growth of CNE-2 cell was suppressed(P=0.015) and its apoptosis was triggered(P=0.008).CONCLUSIONS:The results show that the growth of CNE-2 cell is inhibited and the apoptosis is promoted when Pokemon gene is silenced.The mechanism of this phenomenon may related to the up-expression of p14ARF and p53 gene.更多还原