Gebremedhin Senait, Singh Aruna, Koons Stephen, Bernt William, Konopka Krystyna, Duzgunes Nejat
Department of Biomedical Sciences, Arthur A. Dugoni School of Dentistry, University of the Pacific, San Francisco, CA 94115, USA.
Particle Characterization Laboratories, Novato, CA 94945, USA.
Eur J Pharm Sci. 2014 Aug 18;60:72-9. doi: 10.1016/j.ejps.2014.03.003. Epub 2014 Apr 19.
Suicide gene therapy of oral squamous cell carcinoma (OSCC) may be a viable approach to the treatment of this cancer. However, human OSCC cells are relatively resistant to efficient transfection by non-viral vectors. To identify an optimal vector for gene delivery, we compared the transfection activities and efficiencies of Glycofect, Metafectene, Metafectene Pro, Metafectene Easy and FuGENE HD, using the OSCC cell line, HSC-3, and the cervical carcinoma cell line, HeLa. The size distribution and ζ-potential of the complexes of these vectors with plasmid DNA were assessed by nanoparticle tracking analysis and electrophoretic mobility measurements, respectively. Metafectene Easy and FuGENE HD mediated the highest transfection activity (measured as luciferase expression) and efficiency (measured as the percentage of cells transfected with ß-galactosidase). These vectors were used to deliver a plasmid encoding herpes simplex virus thymidine kinase, followed by ganciclovir treatment. By day 9, HeLa cell viability was 22±3% of controls with FuGENE HD and 26±3% with Metafectene Easy. The viability of HSC-3 cells was 42±25% with FuGENE HD, and 58±28% with Metafectene Easy. The reduction in viability was statistically significant in both cases (p⩽0.005; average of 3 independent experiments), although there was considerable variability between experiments with the HSC-3 cells.
口腔鳞状细胞癌(OSCC)的自杀基因疗法可能是治疗这种癌症的一种可行方法。然而,人类OSCC细胞对非病毒载体的有效转染相对具有抗性。为了确定用于基因递送的最佳载体,我们使用OSCC细胞系HSC-3和子宫颈癌细胞系HeLa,比较了Glycofect、Metafectene、Metafectene Pro、Metafectene Easy和FuGENE HD的转染活性和效率。分别通过纳米颗粒跟踪分析和电泳迁移率测量评估了这些载体与质粒DNA复合物的大小分布和ζ电位。Metafectene Easy和FuGENE HD介导了最高的转染活性(以荧光素酶表达衡量)和效率(以用β-半乳糖苷酶转染的细胞百分比衡量)。这些载体用于递送编码单纯疱疹病毒胸苷激酶的质粒,随后进行更昔洛韦处理。到第9天,使用FuGENE HD时HeLa细胞活力为对照的22±3%,使用Metafectene Easy时为26±3%。使用FuGENE HD时HSC-3细胞的活力为42±25%,使用Metafectene Easy时为58±28%。在这两种情况下,活力的降低均具有统计学意义(p⩽0.005;3次独立实验的平均值),尽管在HSC-3细胞的实验之间存在相当大的变异性。