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目的：探讨人端粒酶逆转录酶（hTERT），癌基因蛋白（C-myc）, 存活素（Survivin）, 血管内皮生长因子（VEGF）基因对人类鼻咽癌细胞生长的影响，以及同时编码四个基因的短发夹重组质粒对人鼻咽癌细胞生长抑制作用及机制。方法：利用基因重组技术构建一个同时靶向作用四个基因的短发夹双链RNA（shRNA）真核表达载体和靶向单独作用hTERTmRNA 的shRNA 真核表达载体，脂质体法转染人CNE-2Z 细胞；试验分组：空白对照组BC 组（不进行干扰），阴性对照组NC 组（加入阴性质粒），A 组（hTERT 单基因质粒组），B 组（多基因联合质粒组）。激光共聚焦显微镜观察转染情况；MTT 法检测细胞增殖活性；RT-PCR 和 Western blot 法分别检测转染后细胞内各基因mRNA 和蛋白表达情况。结果：MTT 法检测，与BC 相，NC 组相比，A 组和B 组的细胞增殖活性均降低，与A 组相比，B 组的增殖活性降低更显著；RT-PCR，Western blot 法,A 组和B 组mRNA 和蛋白表达水平均降低，B 组降低更显著。结论：四个基因共同参与了鼻咽癌细胞的发生和发展。多个基因的联合干扰与单基因干扰相比，能更高效下调各基因蛋白在鼻咽癌细胞的表达水平，更好抑制鼻咽癌细胞的增殖。

英文摘要:

Objective: To investigate the effect of hTERT, C-myc Survivin and VEGF on the human nasopharyngeal carcinoma cell growth, and to investigate the mechanism of short hairpin recombinant plasmid targeting four genes having effect on human nasopharyngeal carcinoma cell growth. Methods: RNAi technology was used to construct a short hairpin recombinant plasmid containing four short hairpin RNAS (shRNA) targeting four genes simultaneously while another plasmid just containing single shRNA against hTERTmRNA, and they were transfected into CNE-2Z cells separately by Liposome; Test groups: Blank control group termed BC Group (no interference) , Negative control group termed NC Group (the plasmid without targeted gene), A group (single-gene plasmid against hTERT), B Group (combined plasmid hitting multiple gene); The transfection was detected under the LSCM; Cell viability was detected by using the MTT; The mRNA and protein expression were determined by RT-PCR and western blot-ting. Results: Compared with that in BC group and NC group, the cell proliferation viability decreased both in A group and B, and which was more lower in B group. The mRNA and protein expression level also reduced significantly, and which was more lower in B group. Conclusions: Four genes are involved in the occurrence and development of nasopharyngeal carcinoma. The combined gene therapy has a better effect on the inhibition of Nasopharyngeal carcinoma cell proliferation than the single-gene interference does.

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