摘要
目的:探讨VEGFsiRNA对肝癌SMMC7721细胞株VEGF表达和增殖的抑制作用。方法:采用siRNA设计法则并用计算机辅助设计9条VEGFsiRNA序列,长度为19个核苷酸;体外构建9条VEGFsiRNA序列,通过脂质体介导转入肝癌SMMC7721细胞株,培养72h,采用CCK8试验分析VEGFsiRNA(25nmol/L)对细胞增殖的抑制效果,用ELISA法检测培养液中VEGF蛋白水平,流式细胞仪检测细胞周期的改变。结果:VEGFsiRNA1-VEGFsiRNA9序列对肝癌细胞的增殖有明显抑制作用,与脂质体对照组和空白对照组相比均有显著差异(P<0·05);其中,VEGFsiR-NA2、VEGFsiRNA4、VEGFsiRNA6、VEGFsiRNA7、VEGFsiRNA8、VEGFsiRNA9对细胞增殖的抑制效果均强于反义寡核苷酸组。VEGFsiRNA1-VEGFsiRNA9序列均能下调肝癌细胞VEGF蛋白的表达水平,以VEGFsiRNA7和VEGFsiR-NA2的效果最强,抑制率分别为52·65%和50·43%。VEGFsiRNA将肝癌细胞阻滞在S期,但未引起细胞凋亡现象。结论:成功地设计和合成VEGFsiRNA。后者能有效地抑制肝癌细胞的增殖,降低VEGF蛋白的表达量。
AIM： To determine the inhibition of VEGF small interfering RNA （siRNA） on expression of VEGF protein and SMMC 7721 cell growth. METHODS： Nine VEGFsiRNA sequences with nineteen nucleotides were designed under the assistance of computer. The inhibitory effect of VEGFsiRNA was analyzed by CCK8. The protein level of VEGF in the media was determined by ELISA. The change of cell cycle was detected by flow cytometry. RESULTS： All the VEGFsiRNA were capable of inhibiting the proliferation of SMMC 7721 cells significantly compared with control and lipofectamine control, while the inhibitory effects of VEGFsiRNA2, VEGFsiRNA4, VEGFsiRNA6, VEGFsiRNA7, VEGFsiRNA8 and VEGFsiRNA9 were better than that of antisense oligodeoxynucleotide. All the VEGFsiRNA reduced the expression of VEGF protein. The effect of VEGFsiRNA7 and VEGF- siRNA2 were the best with the inhibitory rates of 52.65% and 50.43%, respectively. VFGFsiRNAs induced the S arrest in SMMC 7721 cells. The signs of apoptosis in SMMC 7721 cells induced by VFX;FsiRNA were not observed. CONCLUSION： VEGFsiRNA sequences were designed and synthesized successfully. VEGFsiRNA effectively inhibited the proliferation of SMMC 7721 cells and reduced the level of VFGF protein.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第4期771-775,共5页
Chinese Journal of Pathophysiology
基金
广东省科技重点基金资助项目(No.2001-Z-037-01)
广东省自然科学基金资助项目(No.021195)
