Transfection and identification of the cloned strain that stably expressing glucagon like peptide-2 receptor in CaCO2 cell lines.
- Author:
Yun ZHAO
1
;
Feng-jun WANG
;
Pei WANG
;
Hua-bing QI
;
Shi-liang WANG
Author Information
- Publication Type:Journal Article
- MeSH: Caco-2 Cells; Cellular Structures; metabolism; Cloning, Molecular; Gene Expression; Genetic Vectors; Glucagon-Like Peptide 2; genetics; metabolism; Glucagon-Like Peptide-2 Receptor; Humans; Receptors, Glucagon; genetics; metabolism; Transfection
- From: Chinese Journal of Burns 2006;22(4):258-261
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish Caco2 cell line with stable expression of glucagon like peptide-2 receptor( GLP-2R) , in order to establish an in vitro model for the study of protective mechanism of GLP-2 of the intestinal tract.
METHODSThe GLP-2R/pcDNA3. 1 ( + ) plasmid was verified by restriction endonuclease and sequencing , and then it was transfected into Caco2 cells with lipofectamine. After G418 selection, the clones with stable expression of GLP-2R were obtained by limited dilution cloning and expanding. The mRNA and protein expression of GLP-2R in normal human intestine, Caco2 cells, HER293, VE cells, as well as in transfected Caco2 cells were determined with RT-PCR and Western blot.
RESULTSThe sequence of GLP-2R/pcDNA 3. 1 plasmid was correct. No expression of GLP-2R mRNA and protein was found in HER293 and VE cells, but weak expression were found in Caco2 cells, and strong expression was found in normal human intestines. The expression of GLP-2R mRNA and protein expression in Caco2/GLP-2R ( + ) cells were obviously increased after transfection.
CONCLUSIONGLP-2R has special distribution. The expression of GLP-2R is weak in normal Caco2 cells. The establishment of Caco2/GLP-2R ( + ) cellular model is beneficial for the further research of the mechanism of action of GLP-2.
