Combination of Pdx1 and NeuroD1 results in Nkx6.1 and GLUT2 expressions in L02 cells
- VernacularTitle:Pdx1与NeuroD1联合诱导L02细胞Nkx6.1和GLUT2的表达
- Author:
Xiaolong TANG
;
Min GUO
;
Yuan ZHANG
;
Kanger ZHU
- Publication Type:Journal Article
- Keywords:
Pancreatic and duodenal homeobox protein 1;
Neurogenic differentiation factor 1;
Plasmids;
Transcription factors
- From:Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a recombinant plasmid vector containing human pancreatic duodenal homeobox1(Pdx1) and neurogenic differentiation 1(NeuroD1) genes,and to detect its effective expression in eukaryocytes and the ability to induce differentiation of hepatocytes.METHODS: Using human embryo pancreas mRNA as template,Pdx1 and NeuroD1 genes were amplified by RT-PCR and cloned into the two different multiple cloning sites(MCSA and MCSB) of plasmid pIRES.The recombinant plasmid pI/Pdx1/NeuroD1 was transfected into L02 cells.The expression of Pdx1 and NeuroD1 in transfected cells was detected by immunocytochemistry,IFA,RT-PCR and Western blotting,respectively.RESULTS: The length and sequence of cloned segments were correct.Pdx1 and NeuroD1 were expressed in eukaryocytes.Furthermore,the hepatic cells were induced to express glucose transporter 2(GLUT2) and eukaryocyte transcription factor Nkx6.1,which were functionally correlated to ? cells.CONCLUSION: pI/Pdx1/NeuroD1 plasmid is successfully constructed and expressed in human eukaryocytes,with which the cells express the eukaryocyte transcription factor and GLUT2,indicating the transfected cells functionally correlates to ? cells.The results suggest that Pdx1 and NeuroD1 genes can induce the differentiation the cells from hepatic cells to pancreatic endocrine secretion cells.
