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