转染反义细胞周期蛋白A的舌癌细胞系的建立与鉴定

华西口腔医学杂志

转染反义细胞周期蛋白A的舌癌细胞系的建立与鉴定

周峻,杨连甲,金岩,董绍忠,赵宇

710032 第四军医大学秦都口腔医学院病理科

收稿日期:2001-10-25 修回日期:2001-10-25 出版日期:2001-10-20 发布日期:2001-10-20

Establishment and Identification of Squamous Cell Lines Carcinoma of Tongue Transferred with Anti-sense Cyclin A Gene

Zhou Jun, Yang Lianjia, Jin Yan, et al

Department of Pathology,Qindu Stomatological College,the Fourth Military University

Received:2001-10-25 Revised:2001-10-25 Online:2001-10-20 Published:2001-10-20

摘要/Abstract

摘要：

目的:研究反义细胞周期蛋白(cyclin A)基因对舌癌细胞周期的调控作用,为cyclin A在基因治疗方面提供理论基础。方法:采用阳离子脂质体转染方法将已构建成功含人全长cyclinA(1177 kb)的反义真核表达载体pAS-A导入舌癌细胞系(Tca8113),经G418选择性培养液筛选,获得阳性克隆细胞,通过细胞原位杂交检测cyclin A基因的稳定转染情况。结果:pAS-A成功转染进入Tca8113,转染后细胞内cyclin A mRNA的转录和蛋白表达明显降低。结论:反义cyclin A基因稳定有效的在Tca8113细胞系中表达。

关键词: 细胞周期蛋白A, 反义技术, 基因转染

Abstract:

Objective:The aim of this study is to investigate the relationship between cyclinAand cycle regulation of squamous cells car- cinoma of tongue, and to provide basisfor cancergene therapy.Methods: Eukaryocyte expression vector(pAS-A) containing anti- sense and the full-length human cyclin A complementary DNA (cDNA) (1.77 kb) was constructed and was transferred into squa- mous carcinoma of the tongue cell line (Tca8113) by LipofectAMINETMintroduction. The positive cell cloneswere selected with G418. Transcription of Neo gene mRNA and cyclin A mRNA were determined by in situ hybridization. The stable expression of anti-sense cyclin A in the Tca8113 cell line was determined using immunohistochemical methods.Results: After G418 selection, cells transferred anti-sense cyclin Awere obtained successfully. The positive cells of in situ hybridization of specific-stained Neo- probe were observed in cells transferred eukaryocyte expression vector. The positive cells of in situ hybridization of cyclin A in cells transferred anti-sense cyclin Awere significantly fewer than those in cells transferred cyclin A. The positive rate of cyclin A immunohistochemical stain in cells transferred anti-sense cyclin A was significantly lower than that in cells transferred cyclin A. Conclusion:Human anti-sense cyclin A gene is stably expressed in the Tca8113 cell lines.

Key words: cyclin A, antisense, gene transfection

周峻,杨连甲,金岩,董绍忠,赵宇. 转染反义细胞周期蛋白A的舌癌细胞系的建立与鉴定[J]. 华西口腔医学杂志.

Zhou Jun, Yang Lianjia, Jin Yan, et al. Establishment and Identification of Squamous Cell Lines Carcinoma of Tongue Transferred with Anti-sense Cyclin A Gene[J]. West China Journal of Stomatology.

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