In vitro experimental study of killing Hep-2 cells of laryngeal cancer with suicide fusion gene CDglyTK / 临床耳鼻咽喉头颈外科杂志
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
;
(24): 555-558, 2007.
Artigo
em Chinês
| WPRIM
| ID: wpr-748372
ABSTRACT
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Timidina Quinase
/
Proteínas Recombinantes de Fusão
/
Terapia Genética
/
Neoplasias Laríngeas
/
Linhagem Celular Tumoral
/
Citosina Desaminase
/
Genes Transgênicos Suicidas
/
Vetores Genéticos
/
Genética
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
Ano de publicação:
2007
Tipo de documento:
Artigo
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