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脂质体介导的基因转移至头颈癌细胞系的体外研究。

In vitro studies of liposome-mediated gene transfer into head and neck cancer cell lines.

作者信息

Wollenberg B, Lang S, Schmitt B, Kastenbauer E, Zeidler R

机构信息

Department of Oto-Rhino-Laryngology, University of Munich, Germany.

出版信息

Eur Arch Otorhinolaryngol. 1997;254 Suppl 1:S130-2. doi: 10.1007/BF02439743.

Abstract

The 5-year survival rate of patients with squamous cell carcinoma of the head and neck (HNSCC) has remained poor despite innovative surgery and new radiation and chemotherapeutic strategies. In such patients, gene therapy relying on the modification of tumor cells by gene transfer may have great potential as a new treatment modality in the therapy of HNSCC. In the present study we developed an in vitro model to show the efficacy and technical feasibility of cationic liposome-mediated gene transfer into HNSCC. Five adherent squamous cell carcinoma cell lines were transfected with SV40- or CMV-promoter-driven CAT (chloramphenicol-acetyl-transferase)-expression plasmids using DOTAP as the liposome carrier. The level of CAT expression was shown to correlate directly with the amount of transfected DNA and could be measured by a CAT-enzyme-linked immunosorbent assay. The results of gene transfer by liposome-DNA complexes obtained for all cell lines showed a dose-dependent efficacy correlating to the amount of DOTAP employed. The data demonstrate the successful in vitro transfection of epithelial cell lines with DNA, suggesting its usefulness as a new tool for head and neck cancer therapy in vivo.

摘要

尽管有创新的手术以及新的放疗和化疗策略,头颈部鳞状细胞癌(HNSCC)患者的5年生存率仍然很低。对于这类患者,通过基因转移修饰肿瘤细胞的基因治疗作为HNSCC治疗的一种新方法可能具有巨大潜力。在本研究中,我们建立了一个体外模型,以展示阳离子脂质体介导的基因转移至HNSCC的有效性和技术可行性。使用DOTAP作为脂质体载体,用SV40或CMV启动子驱动的氯霉素乙酰转移酶(CAT)表达质粒转染五种贴壁鳞状细胞癌细胞系。结果显示,CAT表达水平与转染DNA的量直接相关,并且可以通过CAT酶联免疫吸附测定法进行检测。所有细胞系通过脂质体-DNA复合物进行基因转移的结果显示出与所用DOTAP量相关的剂量依赖性效果。数据表明DNA成功地在体外转染了上皮细胞系,表明其作为体内头颈部癌症治疗新工具的有用性。

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