Effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of β-catenin in HT-29 cells
10.3969/j.issn.1672-7347.2010.02.009
- VernacularTitle:APC不同功能区域对结肠癌细胞株HT-29中β-连环蛋白表达的影响
- Author:
Liang Lü
;
Jirong HUO
;
Jia LIU
;
Jie WU
;
Jie WANG
- Publication Type:Journal Article
- Keywords:
adenomatous polyposis coli;
recombinant expressive vector;
transfection;
β-catenin
- From:
Journal of Central South University(Medical Sciences)
2010;35(2):140-145
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of β-catenin in human colorectal cancer cells HT-29.Methods The recombinant plasmids were transfected into HT-29 cells mediated by lipofectamine~(TM) 2000, and detected by green fluorescence and RT-PCR. Western blot was applied to detect β-catenin expression level in HT-29 cells after transfection, and gray scales of electrophoresis strips were analyzed by SPSS 13.0.Results Green fluorescence and RT-PCR made clear that all 5 recombinant plasmids were successfully expressed in HT-29 cells. Western blot showed that β-catenin expression level in HT-29 cells was not affected after being transfected with pEGFP-N3-APC1, pEGFP-N3-APC2 and pEGFP-N3-APC3, and was distinctly affected after being transfected with pEGFP-N3-APC4 and pEGFP-N3-APC5, especially the later one. Conclusion The selected APC5 gene fragment with 15-amino acid repeats and SAMP repeats, which is relatively short, can degrade β-catenin level in HT-29 cells and may be applied in the gene therapy.
