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通过粒子轰击对正常和恶性口腔上皮进行转导。

Transduction of normal and malignant oral epithelium by particle bombardment.

作者信息

Shillitoe E J, Noonan S, Hinkle C C, Marini F C, Kellman R M

机构信息

Department of Microbiology and Immunology, State University of New York College of Medicine, Syracuse 13210, USA.

出版信息

Cancer Gene Ther. 1998 May-Jun;5(3):176-82.

PMID:9622101
Abstract

Although genetic approaches to the treatment and prevention of oral cancer are being developed, there are no suitable methods of transduction of the oral mucosa or early cancers. We therefore tested the technique of particle bombardment for its ability to transduce oral cancer cells in vitro and normal epithelium of the hamster cheek pouch in vivo. A gene gun was used to transfer a plasmid that encoded a marker/suicide fusion gene, beta-galactosidase-thymidine kinase (GAL-TEK), under control of a CMV promoter. For comparison we used the method of lipofection and an adenovirus vector. Particle bombardment transduced up to 13% of cells in culture, resulting in a 24.3% reduction in growth in the presence of ganciclovir. The efficiency of transduction was similar to that of lipofection but was much less than that of the adenovirus vector, which transduced 54% of cells and completely inhibited their growth in the presence of ganciclovir. Transduction of the hamster cheek pouch by particle bombardment produced expression of beta-galactosidase as judged by macroscopic staining, for up to 5 days. However, histological examination showed that the transduced cells were rare and superficial, and that administration of systemic ganciclovir did not lead to any changes in the tissue. Improvements in efficiency are necessary before the gene gun can be used in the management of oral cancer.

摘要

尽管针对口腔癌治疗和预防的基因方法正在研发中,但目前尚无适用于口腔黏膜或早期癌症转导的方法。因此,我们测试了粒子轰击技术在体外转导口腔癌细胞以及在体内转导仓鼠颊囊正常上皮细胞的能力。使用基因枪转移一种质粒,该质粒在巨细胞病毒(CMV)启动子的控制下编码一种标记/自杀融合基因,即β-半乳糖苷酶-胸苷激酶(GAL-TEK)。为作比较,我们使用了脂质体转染法和腺病毒载体。粒子轰击在培养物中转导了高达13%的细胞,在更昔洛韦存在的情况下,细胞生长减少了24.3%。转导效率与脂质体转染法相似,但远低于腺病毒载体,腺病毒载体转导了54%的细胞,并在更昔洛韦存在的情况下完全抑制了它们的生长。通过粒子轰击对仓鼠颊囊进行转导,通过宏观染色判断,β-半乳糖苷酶的表达可持续长达5天。然而,组织学检查显示,转导的细胞很少且位于表层,全身给予更昔洛韦并未导致组织发生任何变化。在基因枪可用于口腔癌治疗之前,提高效率是必要的。

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