微球包载携有HSV-TK基因腺病毒肝动脉栓塞治疗的实验研究  

Experimental Investigation of Hepatic Arterial Embolism Treatment with Microsphere Encapsulation of Adenovirus with HSV-TK Gene

作　　者：刘自明[1] 李馨筱[1] 严律南[1] 

机构地区：[1]四川大学华西医院普外科,成都610041

出　　处：《四川大学学报（医学版）》2009年第3期462-465,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30170925)资助

摘　　要：目的探讨用微球包载携有目的基因腺病毒进行肝动脉栓塞治疗以达到治疗基因对原发性肝癌细胞靶向感染的可行性。方法细菌内同源重组法制备携带HSV-TK基因的腺病毒,扩增提纯获得高滴度的重组腺病毒,用挥发溶媒聚合法制载重组病毒PLG微球。建立大鼠移植性肝癌模型,实验大鼠随机分为对照组、生理盐水组、AdEasy-GFP-TK病毒液组、空白微球组及载AdEasy-GFP-TK病毒微球组5组,经肝动脉插管注射不同物质。除对照组外,注射后均结扎肝固有动脉,继之腹腔注射丙氧鸟苷(GCV)。观察各组的肿瘤体积变化及生存期。原位杂交确定HSV-TK基因的体内转染情况。结果载AdEasy-GFP-TK病毒微球组肿瘤体积显著小于其它组,肿瘤生长率显著低于其它各组,平均生存时间显著长于其它各组。原位杂交在AdEasy-GFP-TK病毒液组和AdEasy-GFP-TK病毒微球组肿瘤组织内查见阳性细胞,其它组的肿瘤组织中均未发现阳性细胞。结论肝动脉内注射载病毒微球,治疗基因能靶向、高效的转染到肝癌细胞并有效表达,从而大大提高基因治疗效果。Objective To evaluate the suitability of the biodegradable microsphere encapsulation of adenovirus as a targeting vector for gene therapy of hepatocellular carcinoma. Methods Technique of homologous recombination in bacteria was applied to generate recombinant adenovirus with HSV-TK gene. After obtained the resulting recombinant adenovirus, the solution evaporation method was applied to encapsulate the recombinant adenovirus in poly （lactie/glyeolic acid） （PLGA） eopolymer. The Wistar rat implantation hepatoeellular carcinoma model was set up by inserting the W256 tumor solid piece into Wistar rat＇s liver. Seven days after implantation on liver, the rats were injected through the proper hepatic artery. All of the rats were divided randomly into five groups： control group（Z1 group） ; normal saline group（Z2 group） ; AdEasy-GFP-TK group（Z3 group） ; placebo PLG microspheres group（Z4 group）; PLG mierospheres encapsulation of AdEasy-GFP-TK group（Z5 group）. All of the proper hepatic arteries were ligated except the rats in Z1 group, following by injecting GCV intraperitoneously. The volume and the weight changes of tumor and the life time of the rats were measured. The in situ hybridization was performed to determinate the HSV-TK gene＇s transfection. Results The growth inhibition of tumor and the mean life length in Z5 group was superior to other groups. The expression of HSV-TK was observed in Z3 group and Z5 group by hybridization in situ. Conclusion Genes for gene therapy could transfected into cells of hepatocellular carcinoma efficiently through the proper hepatic artery injection with recombinant adenovirus encapsulated in PLG mierosphere. This could improve the effect of gene therapy remarkably.

关 键 词：腺病毒载体 肝癌 基因治疗 细菌 微球 

分 类 号：R735.7[医药卫生—肿瘤]