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Anti-tumor Function of Double-promoter Regulated Adenovirus Carrying SEA Gene, in the Treatment of Bladder Cancer

  • Translational Biomedical Research
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Abstract

To construct an adenovirus carrying SEA gene and regulated by telomerase reverse transcriptase (hTERT) and hypoxia-inducible factor (HIF) promoters and investigate its anti-tumor function in vitro, as well as its role in lymphocyte production. hTERT and HIF genes were cloned into adenovirus E1A and E1B shuttle plasmids. The control vector for SEA gene expression is under the regulation of CMV and SV40 promoters. Double regulation was obtained through homologous recombination. The positive clones of replicable adenovirus H2-SEA-Ad were selected by plaque assay. The adenovirus was purified, titrated, and DNA was verified by PCR. The obtained virus was used to infect EJ bladder tumor cells and the SEA Mrna, and protein expression was measured by RT-PCR, western blot, and immunofluorescence microscopy, respectively. Co-culture of lymphocytes and tumor cells was observed dynamically under microscope. The construction of shuttle plasmid p315-CSS-SEA was confirmed by PCR and DNA sequencing. Insertion of superantigen SEA gene in adenovirus (H2-SEA-Ad.SEA) was obtained by homologous recombination. In lymphocytes and tumor cell co-culture, the number of viable tumor cells in test groups was significantly lower than that in control group after 12, 24, and 48 h of treatment. Production of interleukin-2, interleukin-4, and tumor necrosis factor were higher in test groups than in control group. Expression of SEA gene in bladder tumor cells by adenoviral vector caused reduced tumor cell proliferation, as well as stimulation of inflammatory cytokine productions in co-cultures with lymphocytes.

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Authors and Affiliations

  1. Department of Urology, People’s Hospital Affiliated to Jiangsu University, Zhenjiang, 212002, China

    Jianpeng Hu

  2. Department of Urology, The Sun Yet-sen Memorial Hospital Affiliated to Sun Yet-sen University, Guangzhou, 510120, China

    Xujun Xuan

  3. The Affiliated School Of Clinical Medicine, Xuzhou Medical College, Xuzhou, 221009, China

    Conghui Han, Lin Hao, Peiying Zhang, Houguang He, Tao Fan & Binzheng Dong

  4. School of Clinical Medicine, Southeast University, Nanjing, 210009, China

    Meng Chen

  5. Department of Urology, Xuzhou Hospital Affiliated to Southeast University, Xuzhou, 221009, China

    Meng Chen

Authors
  1. Jianpeng Hu
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  2. Xujun Xuan
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  3. Conghui Han
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  4. Lin Hao
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  5. Peiying Zhang
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  6. Meng Chen
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  7. Houguang He
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  8. Tao Fan
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  9. Binzheng Dong
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Corresponding author

Correspondence to Conghui Han.

Additional information

X.Xuan is the co-first author. Xujun Xuan and Jianpeng Hu had equal contribution to this study.

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Hu, J., Xuan, X., Han, C. et al. Anti-tumor Function of Double-promoter Regulated Adenovirus Carrying SEA Gene, in the Treatment of Bladder Cancer. Cell Biochem Biophys 62, 353–359 (2012). https://doi.org/10.1007/s12013-011-9289-y

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  • DOI: https://doi.org/10.1007/s12013-011-9289-y

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