Comparison of hPepT1 transfected MDCK cells to hPepT1 transfected HeLa cells
10.3969/j.issn.1001-1978.2017.02.025
- VernacularTitle:人寡肽转运体1转染 MDCK 细胞与转染 HeLa 细胞的比较研究
- Author:
Min LUO
;
Xiaozhong FU
;
Tao XIAO
;
Wenzheng ZHANG
;
Jing LI
;
Ya CHEN
;
Ting LIU
- Keywords:
MDCK cells;
HeLa cells;
human peptide trans-porter 1;
stable transfection;
real time fluorescent quantitative RT-PCR;
Western blot;
Glysar uptake
- From:
Chinese Pharmacological Bulletin
2017;33(2):280-284
- CountryChina
- Language:Chinese
-
Abstract:
Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.
