靶向超声造影剂促进反义基因转染的体外研究

摘要

目的 探讨超声破坏造影剂微泡联合半乳糖化多聚赖氨酸(G-PLL)对基因治疗肝癌的靶向促进作用.方法 将合成的超声造影剂微泡、G-PLL及c-myc反义寡核苷酸(ASODN)耦联物,结合超声辐照作用于表达c-myc基因的人肝癌及人肺癌细胞株进行体外效应研究.观察细胞形态学改变及耦合物与细胞结合情况并检测c-myc基因的表达情况.结果 镜下显示肝癌细胞较肺癌细胞更易于与耦联物结合,多数细胞呈凋亡改变.经超声辐照,耦联物作用后的人肝癌及肺癌细胞c-myc基因表达量均有降低,而G-PLL对肝癌细胞c-myc基因表达有明显抑制作用.结论 超声辐照下,造影剂结合G-PLL及反义基因耦联物可靶向性促进反义基因转染肝癌细胞.%Objective To investigate the improved effects of ultrasound irradiation microbubble agent conjugating galactose-terminal glyco-poly-l-lysine（G-PLL） and antisense oligodeoxynucleotides(ASODN) in the treatment of hepatoma.Methods We established SMMC-7721 hepatoma cells model and A-549 pulmonary carcinoma cells model both of which had c-myc gene expression.G-PLL was bound to microbubble and c-myc ASODN.Different compounds were given to respective experimental groups and soon were treated with ultrasound probe irradiation.We observed every group under powerful microscope every day.C-myc mRNA was detected by RT-PCR after 48 hours.Results After 48 hours transfection and observation under fluorescence microscope,the SMMC-7721 cell had conside rable fluorescent material on the surface of the cells.Between experimental groups,SMMC-7721 cells group had a significant difference,but there was no significant difference in A549 group.Conclusions Ultrasound irradiation,the compounds of microbubble agent,G-PLL and ASODN have targeted inhibitive effect on the c-myc gene expression of SMMC-7721 hepatoma cells.