Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma

Li,Jun-Wu; Gong,Jun-Yuan; Li,Ke; Liu,Yan; Ye,Qiu-Ping; Liu,Xin; Su,Ze-Xuan

doi:10.3892/ol.2010.230

March-April 2011 Volume 2 Issue 2

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March-April 2011 Volume 2 Issue 2

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Article

Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma

Authors:
- Jun-Wu Li
- Jun-Yuan Gong
- Ke Li
- Yan Liu
- Qiu-Ping Ye
- Xin Liu
- Ze-Xuan Su
View Affiliations / Copyright

Affiliations: Department of Microbiology and Immunology, Medical College, Guangzhou, P.R. China, Department of Urology, The First Affiliated Hospital of Jinan University, West No. 601, Huangpu Road, Guangzhou 510632, P.R. China
Pages: 343-347
|
Published online on: December 30, 2010

https://doi.org/10.3892/ol.2010.230
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Abstract

This study aimed to construct a eukaryotic expression plasmid containing the G250/MN/CA IX (G250) and human granulocyte-macrophage colony stimulating factor (hGM-CSF) genes, and to detect the expression of these proteins in vitro by recombinant plasmids in eukaryotic cells. pORF-hGM-CSF and pcDNA3.0-G250 were used as the template to amplify G250 and hGM-CSF by routine polymerase chain reaction (PCR). The two PCR products were cloned into the eukaryotic vector pVAX1, in order to construct a recombinant plasmid pVAX1-G250-hGM, and the plasmid was transfected into human embryonic kidney 293 cells. The protein expression was then determined by immunocytochemi- stry, atomic force microscopy, ELISA and Western blotting. DNA sequencing showed that the cloned G250 and hGM-CSF sequences were consistent with the reported Gene Bank ones. Moreover, a high expression was noted following recombinant plasmid transfection of the G250 and hGM-CSF proteins. Thus, the eukaryotic expression vector pVAX1-G250-hGM containing G250 and hGM-CSF was constructed, allowing for the investigation of the anti-G250 antigen vaccine and immune response mechanisms of biological immunotherapy in renal cell carcinoma.

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1	Kirkali Z, Tuzel E and Mungan MU: Recent advances in kidney cancer and metastatic disease. BJU Int. 88:818–824. 2001. View Article : Google Scholar : PubMed/NCBI
2	Ljungberg B, Hanbury DC, Kuczyk MA, Merseburger AS, Mulders PF, Patard JJ and Sinescu IC: European Association of Urology Guideline Group for renal cell carcinoma. Eur Urol. 51:1502–1510. 2007. View Article : Google Scholar : PubMed/NCBI
3	Bleumer I, Oosterwijk E and De Mulder PF: Immunotherapy for renal cell carcinoma. Eur Urol. 44:65–75. 2003. View Article : Google Scholar : PubMed/NCBI
4	Van Poppel H, Joniau S and van Gool SW: Vaccine therapy in patients with renal cell carcinoma. European Urology. 55:1333–1344. 2009.PubMed/NCBI
5	Potter CP and Harris AL: Diagnostic, prognostic and therapeutic implications on carbonic anhydrases in cancer. Br J Cancer. 89:2–7. 2003. View Article : Google Scholar : PubMed/NCBI
6	Opavský R, Pastoreková S, Zelník V, Gibadulinová A, Stanbridge EJ, Závada J, Kettmann R and Pastorek J: Human MN/CA9 gene, a novel member of the carbonic anhydrase family: structure and exon to protein domain relationships. Genomics. 33:480–487. 1996.PubMed/NCBI
7	Oosterwijk E, Ruiter DJ, Hoedemaeker PJ, Pauwels EK, Jonas U, Zwartendijk J and Warnaar SO: Monoclonal antibody G250 recognizes a determinant present in renal cell carcinoma and absent from normal kidney. Int J Cancer. 38:489–494. 1986. View Article : Google Scholar : PubMed/NCBI
8	Vissers JL, De Vries IJ, Schreurs MW, Engelen LP, Oosterwijk E, Figdor CG and Adema GJ: The renal cell carcinoma-associated antigen G250 encodes a human leukocyte antigen (HLA)-A2.1-restricted epitope recognized by cytotoxic T lymphocytes. Cancer Res. 59:5554–5559. 1999.
9	Dorai T, Sawczuk IS, Pastorek J, Wiernik PH and Dutcher JP: The role of carbonic anhydrase IX overexpression in kidney cancer. Eur J Cancer. 41:2935–2947. 2005. View Article : Google Scholar : PubMed/NCBI
10	Cruciani M, Mengoli C, Serpelloni G, Mazzi R, Bosco O and Malena M: Granulocyte macrophage colony-stimulating factor as an adjuvant for hepatitis B vaccination: a meta-analysis. Vaccine. 25:709–718. 2007. View Article : Google Scholar : PubMed/NCBI
11	Miyashita T, Shah FA, Marti G, Wang J, Armstrong T, Bonde P, Gibson MK, Yoshimura K, Montgomery EA, Duncan MD, Jaffee EM and Harmon JW: Vaccine impedes the development of reflux-induced esophageal cancer in a surgical rat model: efficacy of the vaccine in a pre-Barrett's esophagus setting. J Gastrointest Surg. 12:2–9. 2008. View Article : Google Scholar : PubMed/NCBI
12	Tani K, Azuma M, Nakazaki Y, et al: Phase I study of autologous tumor vaccines transduced with the GM-CSF gene in four patients with stage IV renal cell cancer in Japan: clinical and immunological findings. Mol Ther. 10:799–816. 2004. View Article : Google Scholar : PubMed/NCBI
13	Schwaab T, Tretter CP, Gibson JJ, Cole BF, Schned AR, Harris R, Wallen EM, Fisher JL, Waugh MG, Truman D, Stempkowski LM, Crosby NA, Heaney JA and Ernstoff MS: Immunological effects of granulocyte-macrophage colony-stimulating factor and autologous tumor vaccine in patients with renal cell carcinoma. J Urol. 171:1036–1042. 2004. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Construction of a fusion expression plasmid containing the G250 gene and human granulocyte-macrophage colony stimulating factor and its significance in renal cell carcinoma

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