Tumor-directed gene therapy in mice using a composite nonviral gene delivery system consisting of the piggyBac transposon and polyethylenimine.
Tumor-directed gene therapy in mice using a composite nonviral gene delivery system consisting of the piggyBac transposon and polyethylenimine.
复制标题
使用由piggyBac转座子和聚乙烯亚胺组成的复合非病毒基因传递系统对小鼠进行肿瘤定向基因治疗
DOI：
10.1186/1471-2407-9-126
复制
发表时间：
2009-04-27
期刊：
影响因子：
3.8
通讯作者：
Xu C
中科院分区：
文献类型：
--
作者：
Kang Y;Zhang X;Jiang W;Wu C;Chen C;Zheng Y;Gu J;Xu C
背景
与病毒载体相比，非病毒载体具有更低的免疫原性、更稳定、更安全、更易于复制等优点，可应用于肿瘤基因治疗。然而，非病毒基因传递系统由于其转染率低和转基因表达时间短而没有得到广泛应用，尤其是在体内。开发一种能够支持体内稳定的基因组整合和持续的基因表达的非病毒基因递送系统是人们所希望的。在这里，我们使用了由猪Bac(PB)转座子和聚乙烯亚胺(PEI)组成的复合非病毒基因递送系统在小鼠卵巢肿瘤中进行长期转基因表达。
方法
构建了在PB转座子作用下编码单纯疱疹病毒胸苷激酶(HSV-tk)和单体红色荧光蛋白(MRFP1)的重组载体pB[Act-RFP，HSV-tk]。通过体内PEI系统将其和pBase质粒注射到小鼠卵巢癌移植瘤中。观察HSV-tk/更昔洛韦(GCV)系统对小鼠腹腔注射GCV后的抗肿瘤作用。对肿瘤组织冰冻切片进行组织学分析和原位末端标记法检测。
结果
通过聚合酶链式反应结合限制性内切酶分析，证实构建了真核表达载体。最后一次ppb/TK转染组在荧光显微镜下可观察到mRFP1的表达。接种GCV后，PB/TK组肿瘤体积明显缩小，抑瘤率为81.96%，而TK组为43.07%。组织学分析显示PB/TK组肿瘤组织内有广泛的坏死和淋巴细胞浸润，而对照组肿瘤组织中仅见少量的坏死和淋巴细胞浸润。原位末端标记法检测表明，GCV进入体内后，转基因细胞发生了细胞凋亡。
结论
我们的结果表明，PB转座子与PEI偶联的非病毒基因递送系统可以作为卵巢癌基因治疗的有效工具。
Background
Compared with viral vectors, nonviral vectors are less immunogenic, more stable, safer and easier to replication for application in cancer gene therapy. However, nonviral gene delivery system has not been extensively used because of the low transfection efficiency and the short transgene expression, especially in vivo. It is desirable to develop a nonviral gene delivery system that can support stable genomic integration and persistent gene expression in vivo. Here, we used a composite nonviral gene delivery system consisting of the piggyBac (PB) transposon and polyethylenimine (PEI) for long-term transgene expression in mouse ovarian tumors.
Methods
A recombinant plasmid PB [Act-RFP, HSV-tk] encoding both the herpes simplex thymidine kinase (HSV-tk) and the monomeric red fluorescent protein (mRFP1) under PB transposon elements was constructed. This plasmid and the PBase plasmid were injected into ovarian cancer tumor xenografts in mice by in vivo PEI system. The antitumor effects of HSV-tk/ganciclovir (GCV) system were observed after intraperitoneal injection of GCV. Histological analysis and TUNEL assay were performed on the cryostat sections of the tumor tissue.
Results
Plasmid construction was confirmed by PCR analysis combined with restrictive enzyme digestion. mRFP1 expression could be visualized three weeks after the last transfection of pPB/TK under fluorescence microscopy. After GCV admission, the tumor volume of PB/TK group was significantly reduced and the tumor inhibitory rate was 81.96% contrasted against the 43.07% in the TK group. Histological analysis showed that there were extensive necrosis and lymphocytes infiltration in the tumor tissue of the PB/TK group but limited in the tissue of control group. TUNEL assays suggested that the transfected cells were undergoing apoptosis after GCV admission in vivo.
Conclusion
Our results show that the nonviral gene delivery system coupling PB transposon with PEI can be used as an efficient tool for gene therapy in ovarian cancer.
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影响因子：
6.4
作者：
Dolivet, G;Merlin, JL;Guillemin, F
通讯作者：
Guillemin, F
DOI：
10.1016/j.ejpb.2007.09.008
发表时间：
2008-03-01
影响因子：
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作者：
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通讯作者：
McIvor, R. Scott