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Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer

  • Xu, Rong (Institute of Molecular Medicine, Huaqiao University) ;
  • Guo, Long-Jiang (Institute of Molecular Medicine, Huaqiao University) ;
  • Xin, Jun (Department of Urology, the First Hospital of Quanzhou Affiliated Fujian Medical University) ;
  • Li, Wen-Mao (Institute of Molecular Medicine, Huaqiao University) ;
  • Gao, Yan (Institute of Molecular Medicine, Huaqiao University) ;
  • Zheng, You-Xian (Quanzhou Center for Disease Control and Prevention) ;
  • Guo, You-Hong (Department of Pharmacy, Quanzhou Medical College) ;
  • Lin, Yang-Jun (Department of Pharmacy, Quanzhou Medical College) ;
  • Xie, Yong-Hua (Department of Pharmacy, Quanzhou Medical College) ;
  • Wu, Ya-Qing (Institute of Molecular Medicine, Huaqiao University) ;
  • Xu, Rui-An (Institute of Molecular Medicine, Huaqiao University)
  • Published : 2013.11.30

Abstract

Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

Keywords

Luciferase assay;lung cancer;tumor-specific promoter;gene therapy;tumor biomarker

References

  1. Bafico A, Liu G, Yaniv A, et al (2001). Novel mechanism of Wnt signalling inhibition mediated by Dickkopf-1 interaction with LRP6/Arrow. Nat Cell Biol, 3, 683-6. https://doi.org/10.1038/35083081
  2. Bao R, Connolly DC, Murphy M, et al (2002). Activation of cancer-specific gene expression by the survivin promoter. J Natl Cancer Inst, 94, 522-8. https://doi.org/10.1093/jnci/94.7.522
  3. Benlloch S, Galbis-Caravajal JM, Alenda C, et al (2009). Expression of molecular markers in mediastinal nodes from resected stage I non-small-cell lung cancer (NSCLC): prognostic impact and potential role as markers of occult micrometastases. Ann Oncol, 20, 91-7.
  4. Calabrese F, Lunardi F, Giacometti C, et al (2008). Overexpression of squamous cell carcinoma antigen in idiopathic pulmonary fibrosis: clinicopathological correlations. Thorax, 63, 795-802. https://doi.org/10.1136/thx.2007.088583
  5. Chen JS LJ, Shen L, Rau KM (2004). Cancer-specific activation of the survivin promoter and its potential use in gene therapy. Cancer Gene Ther, 11, 740. https://doi.org/10.1038/sj.cgt.7700752
  6. Clark JC, Weaver TE, Iwamoto HS, et al (1997). Decreased lung compliance and air trapping in heterozygous SP-B-deficient mice. Am J Respir Cell Mol Biol, 16, 46-52. https://doi.org/10.1165/ajrcmb.16.1.8998078
  7. D'Cunha J, Corfits AL, Herndon JE, 2nd, et al (2002). Molecular staging of lung cancer: real-time polymerase chain reaction estimation of lymph node micrometastatic tumor cell burden in stage I non-small cell lung cancer--preliminary results of Cancer and Leukemia Group B Trial 9761. J Thorac Cardiovasc Surg, 123, 484-91; discussion 91. https://doi.org/10.1067/mtc.2002.119883
  8. Gill DR, Pringle IA, Hyde SC (2009). Progress and prospects: the design and production of plasmid vectors. Gene Ther, 16, 165-71. https://doi.org/10.1038/gt.2008.183
  9. Greenberg AK, Lee MS (2007). Biomarkers for lung cancer: clinical uses. Curr Opin Pulm Med, 13, 249-55. https://doi.org/10.1097/MCP.0b013e32819f8f06
  10. Hong JA KY, Abdullaev Z, Flanagan PT (2005). Reciprocal binding of CTCF and BORIS to the NY-ESO-1 promoter coincides with derepression of this cancer-testis gene in lung cancer cells. Cancer Res, 65, 7763-74. https://doi.org/10.1158/0008-5472.CAN-05-0823
  11. Huang M, Batra RK, Kogai T, et al (2001). Ectopic expression of the thyroperoxidase gene augments radioiodide uptake and retention mediated by the sodium iodide symporter in non-small cell lung cancer. Cancer Gene Ther, 8, 612-8. https://doi.org/10.1038/sj.cgt.7700354
  12. Jemal A, Siegel R, Xu J, Ward E (2010). Cancer statistics, 2010. CA Cancer J Clin, 60, 277-300. https://doi.org/10.3322/caac.20073
  13. Johnson NA, Chen BH, Sung SY, et al (2010). A novel targeting modality for renal cell carcinoma: human osteocalcin promoter-mediated gene therapy synergistically induced by vitamin C and vitamin D(3). J Gene Med, 12, 892-903. https://doi.org/10.1002/jgm.1516
  14. Kelly DP, Scarpulla RC (2004). Transcriptional regulatory circuits controlling mitochondrial biogenesis and function. Genes Dev, 18, 357-68. https://doi.org/10.1101/gad.1177604
  15. Khoor A, Whitsett JA, Stahlman MT, Halter SA (1997). Expression of surfactant protein B precursor and surfactant protein B mRNA in adenocarcinoma of the lung. Mod Pathol, 10, 62-7.
  16. Levine M, Tjian R (2003). Transcription regulation and animal diversity. Nature, 424, 147-51. https://doi.org/10.1038/nature01763
  17. Liu L, Liao GQ, He P, et al (2008). Detection of circulating cancer cells in lung cancer patients with a panel of marker genes. Biochem Biophys Res Commun, 372, 756-60. https://doi.org/10.1016/j.bbrc.2008.05.101
  18. Lunardi F, Villano G, Perissinotto E, et al (2011). Overexpression of SERPIN B3 promotes epithelial proliferation and lung fibrosis in mice. Lab Invest, 91, 945-54. https://doi.org/10.1038/labinvest.2011.1
  19. Ma SH, Chen GG, Yip J, Lai PB (2010). Therapeutic effect of alpha-fetoprotein promoter-mediated tBid and chemotherapeutic agents on orthotopic liver tumor in mice. Gene Ther, 17, 905-12. https://doi.org/10.1038/gt.2010.34
  20. Mao B, Wu W, Li Y, et al (2001). LDL-receptor-related protein 6 is a receptor for Dickkopf proteins. Nature, 411, 321-5. https://doi.org/10.1038/35077108
  21. Melton KR, Nesslein LL, Ikegami M, et al (2003). SP-B deficiency causes respiratory failure in adult mice. Am J Physiol Lung Cell Mol Physiol, 285, L543-9. https://doi.org/10.1152/ajplung.00011.2003
  22. Osaki T TY, Tachibana I, Hosoe S (1994). Gene therapy for carcinoembryonic antigen-producing human lung cancer cells by cell type-specific expression of herpes simplex virus thymidine kinase gene. Cancer Res, 54, 5258-61.
  23. Plautz GE, Yang ZY, Wu BY, et al (1993). Immunotherapy of malignancy by in vivo gene transfer into tumors. Proc Natl Acad Sci U S A, 90, 4645-9. https://doi.org/10.1073/pnas.90.10.4645
  24. Podolska K, Stachurska A, Hajdukiewicz K, Malecki M (2012). Gene therapy prospects--intranasal delivery of therapeutic genes. Adv Clin Exp Med, 21, 525-34.
  25. Qiao J DM, Sauter BV, Huang Y (2002). Tumor-specific transcriptional targeting of suicide gene therapy. Gene Therapy, 9, 168-75. https://doi.org/10.1038/sj.gt.3301618
  26. Scholz IV, Cengic N, Baker CH, et al (2005). Radioiodine therapy of colon cancer following tissue-specific sodium iodide symporter gene transfer. Gene Ther, 12, 272-80. https://doi.org/10.1038/sj.gt.3302410
  27. Sheng SL, Huang G, Yu B, Qin WX (2009). Clinical significance and prognostic value of serum Dickkopf-1 concentrations in patients with lung cancer. Clin Chem, 55, 1656-64. https://doi.org/10.1373/clinchem.2009.125641
  28. Shi YZ, Zhang J, Liu ZL, et al (2010). Adenovirus-mediated and tumor-specific transgene expression of the sodium-iodide symporter from the human telomerase reverse transcriptase promoter enhances killing of lung cancer cell line in vitro. Chin Med J (Engl), 123, 2070-6.
  29. Shieh JM, Tang YA, Yang TH, et al (2013). Lack of Association of C-Met-N375S Sequence Variant with Lung Cancer Susceptibility and Prognosis. Int J Med Sci, 10, 988-94. https://doi.org/10.7150/ijms.5944
  30. Siegel R, Naishadham D, Jemal A (2013). Cancer statistics, 2013. CA Cancer J Clin, 63, 11-30. https://doi.org/10.3322/caac.21166
  31. Stewart MJ, Plautz GE, Del Buono L, et al (1992). Gene transfer in vivo with DNA-liposome complexes: safety and acute toxicity in mice. Hum Gene Ther, 3, 267-75. https://doi.org/10.1089/hum.1992.3.3-267
  32. Suzuki K, Fueyo J, Krasnykh V, et al (2001). A conditionally replicative adenovirus with enhanced infectivity shows improved oncolytic potency. Clin Cancer Res, 7, 120-6.
  33. Tokieda K, Iwamoto HS, Bachurski C, et al (1999). Surfactant protein-B-deficient mice are susceptible to hyperoxic lung injury. Am J Respir Cell Mol Biol, 21, 463-72. https://doi.org/10.1165/ajrcmb.21.4.3436
  34. Vile RG, Hart IR (1993). In vitro and in vivo targeting of gene expression to melanoma cells. Cancer Res, 53, 962-7.
  35. X Cao MY, Wei RC, Zeng Y (2011). Cancer targeting Gene-Viro-Therapy of liver carcinoma by dual-regulated oncolytic adenovirus armed with TRAIL gene. Gene Therapy, 18, 765-77. https://doi.org/10.1038/gt.2011.16
  36. Yin X, Meng F, Qu W, et al (2013). Clinical, radiological and genetic analysis of a male infant with neonatal respiratory distress syndrome. Exp Ther Med, 5, 1157-60. https://doi.org/10.3892/etm.2013.970
  37. Yin X, Meng F, Wang Y, et al (2013). Surfactant protein B deficiency and gene mutations for neonatal respiratory distress syndrome in China Han ethnic population. Int J Clin Exp Pathol, 6, 267-72.