抗VEGF发夹状核酶基因对人鼻咽癌细胞株VEGF表达及细胞增殖的影响

Inhibition of anti-VEGF hairpin ribozyme gene on the proliferation and the expression of VEGF of human nasopharyngeal carcinomar cell lines

目的 探讨抗血管内皮生长因子发夹状核酶基因对人鼻咽低分化鳞癌细胞株CNE—2的VEGF表达及其细胞增殖的影响。方法 采用脂质体介导法将抗VEGF发夹状核酶基因真核表达载体转染CNE-2细胞；RNA斑点杂交检测核酶基因在CNE-2细胞中的表达；免疫组化、间接免疫荧光染色及流式细胞仪结合免疫荧光检测转染前后CNE-2细胞中VEGF表达；MTT法、细胞贴壁克隆形成实验、软琼脂克隆形成实验及流式细胞仪进行细胞周期分析，观察其对CNE-2细胞增殖的影响。结果 转染后的CNE—2细胞VEGF表达明显降低；细胞生长减慢，细胞贴壁克隆形成及软琼脂克隆形成率明显降低；Go／G1期细胞显著增多，S期细胞显著减少（P〈0．01）。结论 抗VEGF发央状核酶基因可显著抑制CNE-2细胞的VEGF表达和ENE-2细胞增殖。

Objective To study the inhibition of proliferation and the expression of vascular endothelial growth factor （VEGF） of antiVEGF hairpin ribozyme gene on the cultured tumor ceils of human nasopharyngeal careinomar cell lines （CNE-2） in vitro. Methods AntiVEGF hairpin ribozyme gene eukaryotic expression vector was introduced into CNE-2 ceils by lipofection mediation, ribozyme expression was confirmed by RNA dot blot; the VEGF expression of CNE-2 cells before and after transfection were detected by immunohistochemical and immunofluorescence and flow cytometer immunofluoreseence methods; MTT, colony forming, soft agar colony forming, and flow cytometry （FCM） were used to observe the effect of proliferation on CNE-2 ceils. Results VEGF expression decreased significantly in CNE -2 ceils, the growth of transfected CNE-2 ceils were slower, the average colony forming efficiency and soft agar colony forming efficiency of CNE-2 cells decreased distinctly, the CNE-2 cells of Go/G 1 stage increased, while the cells of S stage were reduced （P〈0.01）. Conclusions Anti-VEGF hairpin ribozyme gene can inhibit significantly the proliferation and the expression of VEGF on CNE-2 ceils.