Objective: The aim of this study is to investigate the relationship between cyclin A and cycle regulation of squamous cells carcinoma of tongue, and to provide basis for cancer gene 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 squamous carcinoma of the tongue cell line (Tca8113) by Lipofect AMINETM introduction. The positive cell clones were 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 A were obtained successfully. The positive cells of in situ hybridization of specific-stained Neoprobe were observed in cells transferred eukaryocyte expression vector. The positive cells of in situ hybridization of cyclin A in cells transferred anti-sense cyclin A were 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.