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Effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of beta-catenin in HT-29 cells


, : Effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of beta-catenin in HT-29 cells. Zhong Nan Da Xue Xue Bao. Yi Xue Ban 35(2): 140-145

To explore the effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of beta-catenin in human colorectal cancer cells HT-29. 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 beta-catenin expression level in HT-29 cells after transfection, and gray scales of electrophoresis strips were analyzed by SPSS 13.0. Green fluorescence and RT-PCR made clear that all 5 recombinant plasmids were successfully expressed in HT-29 cells. Western blot showed that beta-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. The selected APC5 gene fragment with 15-amino acid repeats and SAMP repeats, which is relatively short, can degrade beta-catenin level in HT-29 cells and may be applied in the gene therapy.

US$19.90

PMID: 20197613

DOI: 10.3969/j.issn.1672-7347.2010.02.009


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