摘要
目的探讨结合了肝细胞配体的超声造影剂在超声辐照下介导目的基因转染肝癌细胞的可行性，寻找一种新的基因靶向导入方法。方法制备肝细胞表面特异性配体半乳糖基化多聚赖氨酸（G-PLL），并对产物进行红外光谱分析。将cmyc基因反义寡核苷酸（ASODN）、GPLL与SonoVue微泡三者偶联，在超声波介导下，促使cmycASODN转染人肝细胞癌SMMC7721细胞及肺腺癌A549细胞（作对照），实验结束后行半定量逆转录聚合酶链式反应（RTPCR）以观察不同组间cmyc基因的表达情况。结果超声介导微泡破裂可增加c—myc ASODN在SMMC7721细胞和A549细胞中的表达；SMMC-7721细胞加入G—PLL组中c—myc基因表达水平显著降低，差异有统计学意义（P〈0．01），而A-549细胞中加入与未加入GPLL组间ctnyc基因表达量的差异无统计学性意义（P〉0．05）。结论超声介导微泡造影剂破裂的方法可促进基因的转染，而超声联合肝细胞受体介导的做泡造影剂则更具有肝细胞靶向性，能有效地促进目的基因在肝癌细胞中的表达，为肝癌基因治疗提供了新的途径。
Objective To develop a novel approach of transfecting the objective gene tagetedly mediated by ultrasound contrast agent conjugated ligand of hepatocyte with ultrasound irradiation. Methods C-myc ASODN and galactosylation polylysine which was specific ligand of hepatocyte and the surface of SonoVue microbbule were coupled by electrostatic interaction, laver cancer cells SMMC-7721 and lung cancer cells A 549 were transfected with c myc ASODN in vitro by ultrasound-mediated microbubble destruction. The c myc gene expression of SMMC-7721 cells and A 549 cells was detected by semiquantitative RT PCR. Results Ultrasound mediated mierobubble destruction can enhance antisense e-mye gene expression in SMMC-7721 cells and A 549 cells. C myc expression was decreased in transfected SMMC-7721 cells with G PLL（ P 〈0.01）. C myc expression in A-549 cells with G-PLL was similar to without G-PLL （P〉0.05 ）. Conclusions Ultrasound mediated microbubble destruction method can enhance gene transfection, ultrasound and hepatocellular receptor mediated microbubble destruction have target to liver cells, in which the gene expression level was increased. It is a promising method in gene therapy of liver cancer.
出处
《中华超声影像学杂志》
CSCD
北大核心
2009年第10期891-894,共4页
Chinese Journal of Ultrasonography
基金
黑龙江省自然科学基金（D200414）
黑龙江省教育厅科学技术研究项目（10541129）
关键词
超声检查
微气泡
基因疗法
肝肿瘤
Ultrasonography
Microbubbles
Gene therapy
Liver neoplasms
