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Artículo en Chino | WPRIM | ID: wpr-748372

RESUMEN

OBJECTIVE@#To study the killing effect of suicide gene CDglyTK combined with GCV or 5-FC on the human laryngeal carcinoma Hep-2 cell line in vitro.@*METHOD@#Constructed plasmid pcDNA3.1 (-) CMV. CDglyTK was verified by enzyme digestion of Xho I /Hind III and automatic sequence analysis, then it was introduced into Hep-2 cells by electroporation to yield cells expressing CDglyTK stably after selecting with G418(400 ng/L) for 14 da. The expression of CDglyTK mRNA in transfected Hep-2 cells was tested by RT-PCR. Compared with Hep-2 cells transferred with pcDNA3.1(-), in vitro chemosensitivity of CDglyTK-expressing Hep-2 cells to 5-FC, GCV or 5-FC + GCV was detected by MTT assay.@*RESULT@#The recombinant plasmid contained full-length coding region sequence of CD and TK gene. A anticipated 707 bp fragment was amplified from total RNA of CDglyTK-expressing Hep-2 cells by RT-PCR and a fusion protein of 59 000 was detected in cell extract from transfected Hep-2 cells. In vitro study growth of CDglyTK-positive Hep-2 cells were inhibited by 5-FC, GCV or 5-FC + GCV respectively, and the antitumour effect of 5-FC + GCV is superior to 5-FC or GCV.@*CONCLUSION@#CDglyTK may be a candidate for treating human laryngeal cancer.


Asunto(s)
Humanos , Línea Celular Tumoral , Citosina Desaminasa , Genética , Genes Transgénicos Suicidas , Genética , Terapia Genética , Vectores Genéticos , Neoplasias Laríngeas , Genética , Plásmidos , Proteínas Recombinantes de Fusión , Genética , Timidina Quinasa , Genética
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