抗人血管内皮生长因子发夹状核酶对人脐静脉内皮细胞ECV304的影响

Effect of Anti-VEGF Hairpin Ribozyme on ECV304 Cells

目的观察抗人血管内皮生长因子(VEGF)发夹状核酶对人脐静脉内皮细胞ECV304生物学特性的影响。方法将抗人VEGF发夹状核酶(RZ)基因构建的pcDNA3.1+/RZ转染ECV304细胞,经G418筛选后,用RT-PCR法鉴定RZ基因在ECV304细胞中的转录,用MTT法和流式细胞仪检测转染前后ECV304细胞增殖活力和细胞周期,ELISA法检测细胞上清VEGF的表达水平。结果RZ基因在ECV304细胞中获得转录,它不改变细胞增殖活力和细胞周期,但能明显下调VEGF的表达。结论抗人VEGF发夹状核酶能够显著抑制ECV304细胞VEGF的表达,为进一步开展核酶的肿瘤基因治疗提供了实验依据。

Objective To observe the biological effects of anti-VEGF hairpin ribozyme（RZ） on ECV304 cells. Methods The plasmid pcDNA3.1 ＋ constructed by anti-VEGF RZ was transfected into ECV304 cells and the positive clones were screened by G418. The ribozyme gene transcription in ECV304 cells was confirmed by RT-PCR. The proliferation activity and cell cycle of ECV304 cells before and after transfection was inspected by MTT assay and FCAS. Then the VEGF expression was determined by ELISA assay. Results The ribozyme gene can be transcribed successfully in ECV304 cells, which can not change proliferation activity and cell cycle of ECV304 cells, but it can decrease the level of VEGF expression. Conclusion Anti-VEGF hairpin ribozyme gene can inhibit the VEGF expression of ECV304 cells in vitro. This study provided an experimental evidence for further tumor gene therapy of ribozyme.