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.
Article
in Zh
| WPRIM
| ID: wpr-748372
Responsible library:
WPRO
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.
Full text:
1
Index:
WPRIM
Main subject:
Plasmids
/
Thymidine Kinase
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Recombinant Fusion Proteins
/
Genetic Therapy
/
Laryngeal Neoplasms
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Cell Line, Tumor
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Cytosine Deaminase
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Genes, Transgenic, Suicide
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Genetic Vectors
/
Genetics
Limits:
Humans
Language:
Zh
Journal:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
Year:
2007
Type:
Article