论文部分内容阅读
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。
Objective: To investigate the effect of siRNA interference with Pokemon gene on the growth of nasopharyngeal carcinoma CNE-2 cells. METHODS: Three siRNA-encoded DNA sequences were obtained using siRNA prediction software, and synthetic siRNA fragments were prepared. The effect of silencing Pokemon gene was detected by RT-PCR and Western blotting, and the siRNA fragment with the best silencing effect was selected. MTT assay and flow cytometry were used to detect the growth and apoptosis of CNE-2 cells. RESULTS: A siRNA fragment with a ≥50% silencing effect was screened. After transfection of nasopharyngeal carcinoma CNE-2 cells, the expression of Pokemon gene was significantly decreased (P<0.05), and the expression of p14ARF and p53 gene was up-regulated, P<0.05. The proliferation of nasopharyngeal carcinoma CNE-2 cells was inhibited (P=0.015), and the apoptosis rate was increased, P=0.008. Conclusion: Pokemon gene silencing can inhibit the growth of CNE-2 cells and promote apoptosis. The mechanism may be related to the up-regulation of p14ARF and p53 expression.