Recombinant Adeno-Associated Viral Vector Transduction of Human Prostate Cancer Cell Lines

M. B. Batır
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Abstract

Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection (MOI) ratios of 7 rAAV serotypes, AAV 2/1, 2/2, 2/3, 2/5, 2/6, and 2/9, which were expressing the green fluorescent protein (GFP) transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 2/2 and rAAV 2/6 have the most significant ability to transduce PC-3 cells. Although rAAV 2/2 and rAAV 2/6 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20% in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 2/2 and rAAV 2/6 serotypes achieved more than 60% transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20% in DU-145 cells. Overall, this study demonstrated that rAAV 2/2 and rAAV 2/6 could mediate the expression of a transgene with a high transduction efficiency.
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重组腺相关病毒载体转导人类前列腺癌细胞株
摘要:基因治疗的核心是使用病毒载体,即作为输送载体将修复基因输送到靶细胞的工程病毒。因此,本文分析了 7 种不同 rAAV 血清型及其不同数量对难以转染的人类前列腺癌细胞系 PC-3 和 DU-145 的影响。PC-3和DU-145细胞株分别用7种rAAV血清型(AAV 2/1、2/2、2/3、2/5、2/6和2/9)的不同感染倍率(MOI)进行感染,这7种血清型均表达由CMV启动子驱动的绿色荧光蛋白(GFP)转基因。转导效率通过荧光显微镜和流式细胞术进行了分析。rAAV 2/2 和 rAAV 2/6 转导 PC-3 细胞的能力最强。虽然 rAAV 2/2 和 rAAV 2/6 在 DU-145 细胞系中也是转导能力最强的血清型,但在该细胞系中的转导率不超过 20%。另一方面,病毒感染后,PC-3 细胞的存活率与模拟组相比没有差异,而 DU-145 细胞的存活率则显著下降。这项研究确定了 7 种不同 rAAV 血清型对人类癌细胞系的转导效率。rAAV 2/2 和 rAAV 2/6 血清型在 PC-3 细胞中的转导效率超过 60%,但在 DU-145 细胞中的转导效率不超过 20%。总之,这项研究表明,rAAV 2/2 和 rAAV 2/6 能以较高的转导效率介导转基因的表达。
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