Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
권호 표지
DOI QR코드

DOI QR Code

Construction and Expression of an Eukaryotic Expression Vector Containing the IER3 Gene

  • Wang, Zhen (Department of Oncology, Affiliated Hospital of Medical College, Qingdao University) ;
  • Yu, Hong-Sheng (Department of Oncology, Affiliated Hospital of Medical College, Qingdao University) ;
  • Yao, Ru-Yong (Central Laboratory, Affiliated Hospital of Medical College, Qingdao University) ;
  • Qiu, Wen-Sheng (Department of Oncology, Affiliated Hospital of Medical College, Qingdao University) ;
  • Yue, Lu (Department of Oncology, Affiliated Hospital of Medical College, Qingdao University) ;
  • Sui, Ai-Hua (Central Laboratory, Affiliated Hospital of Medical College, Qingdao University) ;
  • Liu, Xiang-Ping (Central Laboratory, Affiliated Hospital of Medical College, Qingdao University) ;
  • Liu, Shi-Hai (Central Laboratory, Affiliated Hospital of Medical College, Qingdao University)
  • Published : 2013.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.

Keywords

References

  1. Akilov OE, Wu MX, Ustyugova IV, et al (2012). Resistance of Sezary cells to TNF-alpha-induced apoptosis is mediated in part by a loss of TNFR1 and a high level of the IER3 expression. Exp Dermatol, 21, 287-92. https://doi.org/10.1111/j.1600-0625.2012.01452.x
  2. Ao L, Liu JY, Gao LH, et al (2008). Differential expression of genes associated with cell proliferation and apoptosis induced by okadaic acid during the transformation process of BALB/c 3T3 cells. Toxicology In Vitro, 22, 116-27. https://doi.org/10.1016/j.tiv.2007.08.013
  3. Arlt A, Schafer H (2011). Role of the immediate early response 3 (IER3) gene in cellular stress response, inflammation and tumorigenesis. Eur J Cell Biol, 90, 545-52. https://doi.org/10.1016/j.ejcb.2010.10.002
  4. Askoxylakis V, Thieke C, Pleger ST, et al (2010). Long-term survival of cancer patients compared to heart failure and stroke: a systematic review. BMC Cancer, 10, 105. https://doi.org/10.1186/1471-2407-10-105
  5. Billmann-Born S, Till A, Arlt A, et al (2011). Genome-wide expression profiling identifies an impairment of negative feedback signals in the Crohn’s disease-associated NOD2 variant L1007fsinsC. J Immunol, 186, 4027-38. https://doi.org/10.4049/jimmunol.1000085
  6. De Luisi A, Ferrucci A, Coluccia AM, et al (2011). Lenalidomide restrains motility and overangiogenic potential of bone marrow endothelial cells in patients with active multiple myeloma. Clin Cancer Res, 17, 1935-46. https://doi.org/10.1158/1078-0432.CCR-10-2381
  7. Han L, Geng L, Liu X, et al (2011). Clinical significance of IEX-1 expression in ovarian carcinoma. Ultrastruct Pathol, 35, 260-6. https://doi.org/10.3109/01913123.2011.608916
  8. Huang YH, Wu JY, Zhang Y, et al (2002). Synergistic and opposing regulation of the stress-responsive gene IEX-1 by p53, c-Myc, and multiple NF-kappaB/rel complexes. Oncogene, 21, 6819-28. https://doi.org/10.1038/sj.onc.1205854
  9. Hu Y, Sun H, Drake J, et al (2004). From mice to humans: identification of commonly deregulated genes in mammary cancer via comparative SAGE studies. Cancer Res, 64, 7748-55. https://doi.org/10.1158/0008-5472.CAN-04-1827
  10. Im HJ, Pittelkow MR, Kumar R (2002). Divergent regulation of the growth-promoting gene IEX-1 by the p53 tumor suppressor and Sp1. J Biol Chem, 277, 14612-21.
  11. Krieger S, Grunau C, Sabbah M, et al (2005). Cyclin D1 gene activation in human myeloma cells is independent of DNA hypomethylation or histone hyperacetylation. Exp Hematol, 33, 652-59. https://doi.org/10.1016/j.exphem.2005.03.011
  12. Lee S, Bang S, Song K, et al (2006). Differential expression in normal-adenoma-carcinoma sequence suggests complex molecular carcinogenesis in colon. Oncol Rep, 16, 747-54.
  13. Lee YH, Kim JH, Zhou H, et al (2012). Salivary transcriptomic biomarkers for detection of ovarian cancer: for serous papillary adenocarcinoma. J Mol Med (Berl), 90, 427-34. https://doi.org/10.1007/s00109-011-0829-0
  14. Matsueda S, Takedatsu H, Sasada T, et al (2007). New peptide vaccine candidates for epithelial cancer patients with HLA-A3 supertype alleles. J Immunother, 30, 274-81. https://doi.org/10.1097/01.cji.0000211340.88835.e7
  15. Santamaria C, Ramos F, Puig N, et al (2012). Simultaneous analysis of the expression of 14 genes with individual prognostic value in myelodysplastic syndrome patients at diagnosis: WT1 detection in peripheral blood adversely affects survival. Ann Hematol, 91, 1887-95. https://doi.org/10.1007/s00277-012-1538-7
  16. Sasada T, Azuma K, Hirai T, et al (2008). Prognostic significance of the immediate early response gene X-1 (IEX-1) expression in pancreatic cancer. Ann Surg Oncol, 15, 609-17. https://doi.org/10.1245/s10434-007-9669-0
  17. Schwander B, Ravera S, Giuliani G, et al (2012). Cost comparison of second-line treatment options for late stage non-small-cell lung cancer: cost analysis for Italy. Clinicoecon Outcomes Res, 4, 237-43.
  18. Segditsas S, Sieber O, Deheragoda M, et al (2008). Putative direct and indirect Wnt targets identified through consistent gene expression changes in APC-mutant intestinal adenomas from humans and mice. Hum Mol Genet, 17, 3864-75. https://doi.org/10.1093/hmg/ddn286
  19. Sipo I, Hurtado Pico A, Wang X, et al (2006). An improved Tet-On regulatable FasL-adenovirus vector system for lung cancer therapy. J Mol Med (Berl), 84, 215-25. https://doi.org/10.1007/s00109-005-0009-1
  20. Steensma DP, Neiger JD, Porcher JC, et al (2009). Rearrangements and amplification of IER3 (IEX-1) represent a novel and recurrent molecular abnormality in myelodysplastic syndromes. Cancer Res, 69, 7518-23. https://doi.org/10.1158/0008-5472.CAN-09-1428
  21. Wang H, Wei F, Zhang J, et al (2012). A novel immunocompetent murine tumor model for the evaluation of RCAd-enhanced RDAd transduction efficacy. Tumour Biol, 33, 1245-53. https://doi.org/10.1007/s13277-012-0374-7
  22. Wu MX (2003). Roles of the stress-induced gene IEX-1 in regulation of cell death and oncogenesis. Apoptosis, 8, 11-8. https://doi.org/10.1023/A:1021688600370
  23. Yang JJ, Preston GA, Alcorta DA, et al (2002). Expression profile of leukocyte genes activated by anti-neutrophil cytoplasmic autoantibodies (ANCA). Kidney Int, 62, 1638-49. https://doi.org/10.1046/j.1523-1755.2002.00619.x
  24. Zilliox MJ, Moss WJ, Griffin DE (2007). Gene expression changes in peripheral blood mononuclear cells during measles virus infection. Clin Vaccine Immunol, 14, 918-23. https://doi.org/10.1128/CVI.00031-07