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Article

Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter

  • Authors:
    • Bin Chen
    • Xiao-Peng Chen
    • Yong Wang
    • Wei Cui
    • Min Zhong
  • View Affiliations / Copyright

    Affiliations: Department of Hepatobiliary Surgery, Affiliated Yijishan Hospital, Wannan Medical College, Wuhu, Anhui 241001, P.R. China, Department of General Surgery, Pukou Traditional Chinese Medicine Hospital, Nanjin 211800, P.R. China, Central Laboratory, Affiliated Yijishan Hospital, Wannan Medical College, Wuhu, Anhui 241001, P.R. China
  • Pages: 41-46
    |
    Published online on: June 7, 2012
       https://doi.org/10.3892/br.2012.1
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Abstract

The transcription activity of the tumor-specific promoter may be increased using specific DNA sequences such as simian virus 40 (SV40). Human heparanase (HPSE) gene promoter is also considered a tumor-specific promoter. However, whether or not the SV40 enhancer affects the tumor specificity of HPSE remains to be determined. The SV40 enhancer sequence, 237 bp in length, was amplified and correctly inserted into the assigned multiple clone sites (MCS) of the eukaryotic expression vector pEGFP-Hp, which was constructed in advance. The recombinant plasmid pEGFP-Hp-SV40e was consistent with the anticipated Genbank data and transfected into human umbilical vein endothelial cell (ECV) and tumor cell lines, including hepatoma carcinoma (HepG2), laryngeal carcinoma (Hep2) and chronic myelogenous leukemia cell lines (K562) using lipofectamine, respectively. The expression of the reporter gene, green fluorescent protein (GFP), was detected using fluorescence microscopy and flow cytometry. The length of the amplified SV40 enchancer was 237 bp and the sequence was in accordance with the GenBank data. The recombinant plasmid pEGFP-Hp-SV40 was consistent with the anticipated results. Fluorimetric analysis showed that the fluorescence of pEGFP-Hp-SV40e in ECV cells was as dim as pEGFP-Hp, and obviously weaker than pEGFP-N1. In tumor cells including HepG2, Hep2 and K562 cells, the fluorescence of pEGFP-Hp-SV40e was similar to that of pEGFP-N1, which was clearly brighter than pEGFP-Hp. The average transfecion rates in the 4 types of cells were 4.1, 17.2, 8.8 and 6.4% in the pEGFP-Hp; 18.3, 29.3, 17.0 and 13.0% in the pEGFP-Nl and 4.3, 28.8, 16.4 and 11.7% in the pEGFP-Hp-SV40e groups, respectively. The ratio of pEGFP-Hp-SV40e to pEGFP-Hp in all cells was 1.05, 1.67, 1.86 and 1.83, respectively. In conclusion, the inserted SV40 enhancer sequence is able to improve the transcriptional activity of the human HPSE gene promoter, but does not affect its tumor specificity.
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Copy and paste a formatted citation
Spandidos Publications style
Chen B, Chen X, Wang Y, Cui W and Zhong M: Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter. Biomed Rep 1: 41-46, 2013.
APA
Chen, B., Chen, X., Wang, Y., Cui, W., & Zhong, M. (2013). Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter. Biomedical Reports, 1, 41-46. https://doi.org/10.3892/br.2012.1
MLA
Chen, B., Chen, X., Wang, Y., Cui, W., Zhong, M."Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter". Biomedical Reports 1.1 (2013): 41-46.
Chicago
Chen, B., Chen, X., Wang, Y., Cui, W., Zhong, M."Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter". Biomedical Reports 1, no. 1 (2013): 41-46. https://doi.org/10.3892/br.2012.1
Copy and paste a formatted citation
x
Spandidos Publications style
Chen B, Chen X, Wang Y, Cui W and Zhong M: Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter. Biomed Rep 1: 41-46, 2013.
APA
Chen, B., Chen, X., Wang, Y., Cui, W., & Zhong, M. (2013). Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter. Biomedical Reports, 1, 41-46. https://doi.org/10.3892/br.2012.1
MLA
Chen, B., Chen, X., Wang, Y., Cui, W., Zhong, M."Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter". Biomedical Reports 1.1 (2013): 41-46.
Chicago
Chen, B., Chen, X., Wang, Y., Cui, W., Zhong, M."Simian virus 40 enhancer does not affect the tumor specificity of human heparanase gene promoter". Biomedical Reports 1, no. 1 (2013): 41-46. https://doi.org/10.3892/br.2012.1
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