In vitro experimental study of killing Hep-2 cells of laryngeal cancer with suicide fusion gene CDglyTK.
- Author:
Yaoyun TANG
1
;
Changning XIE
;
Jianping LIU
;
Suping ZHAO
;
Yongquan TIAN
;
Jianyun XIAO
Author Information
1. Department of Otolaryngology, Hospital of Xiangya, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Cytosine Deaminase;
genetics;
Genes, Transgenic, Suicide;
genetics;
Genetic Therapy;
Genetic Vectors;
Humans;
Laryngeal Neoplasms;
genetics;
Plasmids;
Recombinant Fusion Proteins;
genetics;
Thymidine Kinase;
genetics
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2007;21(12):555-558
- CountryChina
- Language:Chinese
-
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.
