摘要
目的 :试图用计算机设计并筛选出血管内皮生长因子 (VEGF)的高效反义核酸 ;用实验方法研究VEGF反义核酸对K5 6 2细胞增殖的影响。方法 :用RNAstructure (version 3.7)软件 ,选择总自由能 (overall△G3 7)相对低的反义核酸 ,共计 7条 ,长度 18- 2 0核苷酸 ,全硫代修饰 ;细胞培养 72h ,采用台盼蓝拒染法观察存活细胞 ,用ELISA法检测培养液中VEGF蛋白水平 ,分析反义核酸对K5 6 2细胞的作用。结果 :筛选出 6条反义药物对K5 6 2细胞生长有明显抑制作用 ,均优于阳性对照组 (X2 )。与随机对照组 (X1)相比 ,最优序列X7组细胞生长抑制率达31 9%、培养液中VEGF蛋白表达抑制率达 5 1.4 % ,overall△G3 7与反义药物活性密切相关 (r=0 887,P <0 0 1)。结论 :计算机辅助设计有助于获得更好反义药物 ,VEGF反义药物可抑制K5 6 2细胞生长及VEGF蛋白表达 ,内源性VEGF蛋白具有促进K5 6
AIM: The effective antisense sequences targeted VEGF mRNA with computer software would be screened and designed, and effect of them on growth K562 cells and protein expression of VEGF were studied with experiments. METHODS: Seven antisense sequences were selected and synthesized, which consisted of 18-20 deoxynucleotide acid and were modified with phosphorothioate, according to principle of low free energy of overall △G 37 Overall. Cell growth was assayed by trypan blue dye exclusion assay and level of VEGF protien in the media was determined by ELISA. RESULTS: Six of seven sequences were capable of inhibing growth of K562 cells and downregulating the VEGF protein expression significantly, compared with Scrambed control group. It was found that there was a close correlation between low level of overall △G 37 and antisense effectiveness ( r =0 887, P <0 01). CONCLUSION: VEGF mRNA antisense oliogdeoxynucleotides, which were designed by computer software of RNAstructure, were able to inhibit growth of K562 and its protein expression. The VEGF mRNA may be new target attached by drugs. At same time, the computer aided design is useful methods to obtain the effective antisense.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2003年第5期645-648,共4页
Chinese Journal of Pathophysiology
基金
国家攀登计划项目(No.95 -专 - 10 )
广东省自然科学基金重点项目(No.0 2 1195)
广东省科技计划(No .99MO12 0 4G)
广州市科技重点基金资助项目(No.2 0 0 1- 2-0 37- 0 1)
