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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 被引量:3

Generation 5 polyamidoamine dendrimers-mediated experimental HSV-tk gene therapy in murine prostate cancer models
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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.
出处 《中国癌症杂志》 CAS CSCD 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)
关键词 PAMAM-D 基因治疗 自杀基因 前列腺癌 非病毒载体 PAMAM-D gene therapy suicide gene prostate cancer nonviral vector
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参考文献15

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  • 2黄寿吾,王昆,黄复生.新型脂质体的研究进展[J].食品与药品,2005,7(07A):5-9. 被引量:6
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  • 4周少华,陈勇,洪艳.壳聚糖的纳米化及其生物学效应[J].中国组织工程研究与临床康复,2007,11(26):5190-5193. 被引量:3
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  • 6Kang C,Yuan X,Li F,et al.Evaluation of folate-PAMAM for the delivery of antisense oligonucleotides to rat C6 glioma cells in vitro and in vivo[J].J Biomed Mater Res A,2009,93(2):585-594.
  • 7Okegawa T,Li Y,Pong RC,et al.The dual impact of coxsackie and adenovirus receptor expression on human prostate cancer gene therapy[J].Cancer Res,2000,60(18):5031-5036.
  • 8Verma IM,Weitzman MD.Gene therapy:twenty-first century medicine[J].Annu Rev Biochem,2005,74:711-738.
  • 9Choi Y,Thomas T,Kotlyar A,et al.Synthesis and functional evaluation of DNA-assembled polyamidoamne dendrimer clusters for cancer cell-specific targeting[J].Chem Biol,2005,12(1):35-43.
  • 10Wang Y,Kong W,Song Y,et al.Polyamidoamine dendrimers with a modified pentaerythritol core having high efficiency and low cytotoxicity as gene carriers[J].Biomacromolecules,2009,10(3):617-622.

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