沉默Survivin基因对鼻咽癌CNE1细胞增殖和凋亡的研究

目的：探讨 Survivin表达降低时，对鼻咽癌细胞的增殖及凋亡的影响。方法运用 RNA 干扰技术，结合 RT-PCR，Western blot技术，观察干扰 Survivin效果。干扰 Survivn后用 MTT法和 TUNEL法检测鼻咽癌细胞增殖，凋亡的情况。用 Western-blot方法检测凋亡相关蛋白 PARP，Bcl-2和Bax的表达。结果 RT-PCR检测显示 Survivin-siRNA干扰组，Survivin mRNA表达明显下调，其相对表达量为0.26±0.02，抑制率为43.7％，与对照组相比差异具有统计学意义（P＜0.05）；Western blot检测显示 Survivin-siRNA组蛋白表达下调，其蛋白相对表达量下降了57％，与对照组相比差异具有统计学意义（P＜0.05）；MTT检测显示 Survivin-siRNA组对细胞增殖有明显抑制作用，在24，48和72 h的抑制率分别为21.9％，37.1％和29.6％，与对照组相比差异具有统计学意义（P＜0.05）；Tunel染色结果显示 Survivin-siRNA组细胞凋亡数明显增高。Western blot结果显示Survivin-siRNA组凋亡蛋白PARP（89KD）显著升高，为对照组的3.9倍，抑制凋亡蛋白Bcl-2降低，降低了70％，促进凋亡蛋白Bax升高，为对照组的2.4倍；同时也抑制AKT的磷酸化，磷酸化水平降低了57％，与对照组相比以上结果差异均具有统计学意义（P＜0.05）。结论干扰 Survivin表达，抑制鼻咽癌细胞增殖，促进细胞凋亡。Survivin基因可成为鼻咽癌基因治疗的一个潜在靶点。

Research on the Effect of Silence of Survivin Gene by RNAi on the Proliferation and Apoptosis of Nasopharyngeal Carcinoma CNE1 Cells

Objective To explore the effect of decrease of Survivin expression on the proliferation and apoptosis of the naso-pharyngeal carcinoma cells.Methods To observe the effect of decrease of Survivin expression on nasopharyngeal carcinoma cell morphology using RNAi technology combined with RT-PCR and Western blot.Cell proliferation and apoptosis were de-tected by MTT and TUNEL assay.The expression of PARP,Bcl-2 and Bax was detected by Western blot.Results RT-PCR result showed the survivin mRNA expression in Surviving-siRNA groups was downregulated （about 0.26±0.02）and the inhibition ratio was 43.7% （P〈0.05）.MTT result showed cell proliferation rates were significantly different between 24,48 and 72 h after transfection.The cell inhibition rates were 21.9%,37.1% and 29.6%,respectively （P〈0.05）.West-ern blot result showed the Survivin protein expression in Survivin-siRNA2 groups was downregalated and the relative ex-pression level was reduced by 57% （P〈0.05）.TUNEL assay showed that cell apoptosis rate was increased obviously in surviving-siRNA groups.The expression of PARP （89KD）and Bax wasupregulated （3.9 and 2.4 fold change）and the ex-pression of Bcl-2 was downregulated （0.3 fold change）.The phosphorylation of AKT was inhibited when Survivin was down-regulated （about reduced by 5 7%）.Conclusion Silencing Survivin gene by siRNA can inhibit the proliferation of Na-sopharyngeal carcinoma cells and induce apoptosis.Survivin may become a potential gene for therapy target of nasopharynge-al carcinoma.