Abstract:
Objective
To investigate the CEA promoter’s role on CD and TK for gene treatment of tumors.
Methods
Lovo and Hela cells were cultured, passed, stored in liquid nitrogen, transfected with 10 μl recombinant adenovirus with Cp.CD.TK(RA-Cp.C.T),observed the green fluorescence. The total DNA of transfected Lovo and Hela were extracted and employed as PCR templates for amplifing Cp,CD and TK.The three interest genes were identified with agarose electrophoresis.The Lovo and Hela cell transfected with RA-Cp.C.T were overlayed on 96 well cell culture board, and every strain of cell was divided into two groups (24 wells per group). The first group was cultured with growth medium containing 10 mg/L 5-Fc, and the second group with growth medium containing 5 mg/L GCV. Cell morphologic observation with light microscope and cell vitality measure with trypan blue dying were carried out.
Results
All the Lovo and Hela cells,being transfected by RA-Cp.C.T，presented green fluorescence and had Cp,CD and TK DNA identified by PCR.Both 5-Fc and GCV could kill Lovo cells,but almost no influence on Hela cells.In 5-Fc group,the average vitality of Lovo cell was 8.54%±1.34%，while that of Hela cell was 95.05%±2.18%(P＜0.01). In GCV group, the average vitality of Lovo cell was 8.38%±1.22%，while that of Hela cell was 95.39%±1.64%(P＜0.01).
Conclusion
The suicide gene of CD and TK promoted by Cp kill violently CEA positive Lovo cell, and can’t kill CEA negative Hela cell.
Key words:
Human colorectal cancer,
CEA promoter,
Cytosine deaminase,
Thymidine kinase
Tian-bao WANG, Wen-guang DONG, Jian-ping WANG. CEA promoter promoting specifically CD and TK gene’s expression in CEA positive cancer[J]. Chinese Archives of General Surgery(Electronic Edition), 2008, 02(01): 24-27.