靶向 VEGF 基因的 siRNA 体外抑制兔 VX2细胞生长的研究

摘要

Objective:To investigate the effect of small interfering RNA(siRNA)targeting vascular endothelial growth factor(VEGF)on the growth of rabbit squamous epithelial cancer cell line VX2 cells. Methods:Three groups of siRNA targeting VEGF gene in VX2 cells(VEGF - si1,VEGF - si2,VEGF - si3)were designed and obtained by chemical synthesis. Then these siRNAs were transfected into cultured VX2 cells in vitro by Lipofectamine. At 48 hours after VEGF - siRNA transfection,the levels of VEGF mRNA expression were analyzed by reverse transcription poly-merase chain reaction(RT - PCR),the secretion of VEGF protein in the supematant was examined by ELISA,and the viability of VX2 cells was detected by MTT. Results:For VEGF - si1 or VEGF - si3 transfected VX2 cells,The growth of cells was inhibited by the rate of(38. 5 ± 7. 3)% or(30. 0 ± 5. 8)% ,significantly higher than that of scramble siRNA transfected VX2 cells(control group),(P < 0. 01);The level of VEGF mRNA was(0. 37 ± 0. 06)or (0. 45 ± 0. 07),significantly lower than that in control group,(P < 0. 01);The secretion of VEGF protein was also decreased by the rate of(58. 1 ± 7. 3)% or(51. 9 ± 7. 9)% ,notably higher than that of control group,(P < 0. 01). Conclusion:VEGF - targeted siRNA can in vitro specifically inhibit VEGF mRNA transcription,VEGF protein secre-tion and the proliferation of VX2 cell line.%目的：探讨靶向 VEGF 基因的 siRNA 对兔 VX2肿瘤细胞生长的影响。方法：采用化学合成法得到靶向兔 VX2细胞 VEGF 基因的 siRNA 分子，应用脂质体将其转染体外培养的 VX2细胞。转染48h 后，RT - PCR法检测 VEGF mRNA 水平，ELISA 法检测细胞分泌的 VEGF 蛋白，MTT 法检测细胞生长活力。结果：VEGF -si1和 VEGF - si3单独转染 VX2细胞，细胞的生长抑制率分别为（38.5±7.3）％和（30.0±5.8）％，均显著高于 scr - siRNA 转染的对照组（P ＜0.01）；细胞 VEGF mRNA 表达水平分别为（0.37±0.06）和（0.45±0.07），均显著低于对照组（P ＜0.01）；细胞分泌 VEGF 蛋白的抑制率分别为（58.1±7.3）％和（51.9±7.9）％，均显著高于对照组（P ＜0.01）。结论：VEGF siRNA 分子在体外能特异地抑制兔 VX2细胞的 VEGF 基因 mRNA 转录，VEGF 蛋白分泌以及细胞增殖。