新型基因载体G5-PAMAM-D在前列腺癌HSV-tk/GCV自杀基因系统中的应用  

Generation 5 polyamidoamine dendrimers-mediated experimental HSV-tk gene therapy in murine prostate cancer models

作　　者：赵维明[1] 徐勇[1] 王燕铭[2] 程兆康[2] 于琦[2] 孔德领[2] 

机构地区：[1]天津医科大学第二医院泌尿外科天津市泌尿外科研究所,天津300211 [2]南开大学生物活性材料教育部重点实验室,天津300071

出　　处：《中国癌症杂志》2007年第3期225-230,共6页China Oncology

基　　金：教育部高等学校博士学科点专项科研基金资助(基金号:20060062007)

摘　　要：背景与目的:考察纳米级阳离子聚酰胺-胺型树枝状聚合物(polyam idoam ine dendrimers,PAMAM-D)作为基因载体进行前列腺癌自杀基因治疗的可行性,为前列腺癌的基因治疗寻找新的基因载体。方法:以第5代PAMAM-D(G5-PAMAM-D)为载体将含增强型荧光蛋白(EGFP)基因片段的重组质粒pEGFP-C1转染至前列腺癌细胞系PC-3和22Rv1,成功表达EGFP后,再以G5-PAMAM-D为载体将含HSV-tk自杀基因的真核表达重组质粒pcDNA3-tk转染至前列腺癌细胞系PC-3和22Rv1,转染48h后,对转染的两种细胞给予浓度为0、10、100、1000、10000μmol/L前体药更昔洛韦(Ganciclovir,GCV),24h后采用MTT比色法测定药物对细胞增殖的影响。制作前列腺癌皮下荷瘤小鼠,将G5-PAMAM-D/pcDNA3-tk复合物瘤内注射,24h后腹腔注射GCV,观察这一复合物体内对肿瘤生长的抑制作用。结果:荧光观察及FCM结果证明G5-PAMAM-D可将pEGFP-C1转入两种前列腺癌细胞并表达EGFP。采用G5-PAMAM-D将pcDNA3-tk重组质粒转染PC-3和22Rv1细胞,给不同浓度的前体药GCV后,实验组细胞与对照组相比有明显浓度依赖性生长抑制。荷瘤小鼠在给予瘤内注射G5-PAMAM-D/pcDNA3-tk复合物后,行腹腔注射GCV,治疗后第40天,实验组肿瘤体积(1135±245mm3)与对照组裸质粒组(9965±2109mm3)和PAMAM-D组(8357±1956mm3)相比肿瘤生长明显被抑制(P<0.01),生存时间延长。结论:G5-PAMAM-D可作为前列腺腺癌自杀基因治疗的基因载体,有良好的应用前景。Background and purpose： To identify the possibility of Generation 5 Polyamidoamine dendrimers （G5-PAMAM-D） as gene vector for HSV-tk suicide sere therapy in prostate cancer. Methods： Prostrate cancer cells PC-3,22Rv1 were transfected with plasmid pEGFP-C1 coated with G5-PAMAM-D, Furthermore, G5-PAMAM-D were used as gene vectors to deliver plasmid pcDNA3- tk that contain HSV-tk suicide gene into PC-3 and 22Rv1. 48 h after transfection, the precursor ganciclovir（GCV） was added to the transfectod PC-3,22Rv1 cells at fmal concemtration 0, 10,100,1000, 10 000 μmol/L MTT was used to evaluate the effect of GCV on the proliferation of two prostate cancer cells after 24 hour incubation. BALB/c mice were subcutaneously injected with PC-3 that was used as tumor model, Polyplex of G5-PAMAM-D and pcDNA3-tk was injected intratumorally followed by GCV injected intraperitoneally 24 hours later. The tumor volume and mice survival time were observed. Results： G5-PAMAM-D could be used as vector to transfer plasmid pEGFP-C1 into two prostate cancer cells, and EGFP was successfully expressed. The number of live cells in G5-PAMAM- D nanoparficles group with HSV-tk is much less than those of the control group, significant difference was found between those two groups. The tumor volume of BALB/c mice was significantly smaller in PAMAM-D/pcDNA3-tk group（ 1 135 ±245mm^3） than that in Naked plasmid group（9 965 ±2 109mm^3） and PAMAM-D group（8 357 ±1 956mm^3） （P 〈0.01） 40 days after treatment. Survival time of PAMAM-D/pcDNA3-tk group was longer than the two control groups. Conclusions： G5-PAMAM-D nanoparticles can deliver HSV-tk gene to prostate cancer cells effectively; it has possibilities of being used as gene vector for gent therapy in in prostate cancer.

关 键 词：PAMAM-D 基因治疗 自杀基因 前列腺癌 非病毒载体 

分 类 号：R737.25[医药卫生—肿瘤]