Baculovirus Mediated Experimental Research on Targeted Egr1-Kringle 5 Gene Radiotherapy in Lung Adenocarcinoma


Objective: To investigate the feasibility of temporally and spatially restricted Kringle5 expression induced by radiation, as well as the dual effect of radiotherapy and antiangiogenic therapy in lung adenocarcinoma in vitro. Methods: We first constructed recombinant baculovirus vectors containing Egr1 promoter and human plasminogen Kringle5 gene (rhK5), then transfected them into lung adenocarcinoma cells (A549). Transfect efficiency of the baculovirus for gene transfer in A549 cells and the activity of Egr1 promoter induced by X-radiation were detected by fluorescence microscopy. The rhK5 mRNA transcription and rhK5 protein expression were detected by Real-time PCR and Western blot assay, respectively. The apoptosis asssay of human umbilical veins endothelial cells (HUVEC) was analyzed by flow cytometry. Results: The recombinant baculovirus were successfully transfected into A549 and HUVEC cells. As for the temporal regulation, the rhK5 mRNA transcription and rhK5 protein expression were elevated with the irradiation time significantly. And the HUVEC apoptotic percentage increased in relation to the irradiation time as well. As for the spatial regulation, rhK5 mRNA transcription level of A549 cell lines transfected with recombinant baculovirus Egr1-K5 was significantly higher than that of control groups after the same dose of X-radiation. When we analyzed the dose and frequency of X-radiation, no difference was observed among each dose after continuously three-times of irradiation. Conclusion: Baculovirus-mediated Egr1-K5 can be used in gene radiotherapy for its temporary and spatial controllable rhK5 expression by X-radiation and the consequent HUVEC apoptosis in vitro study. And low dose and more times of irradiation might be more effective. It would provide a promising way for the tumor treatment.

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H. Xu, R. Guo and B. Li, "Baculovirus Mediated Experimental Research on Targeted Egr1-Kringle 5 Gene Radiotherapy in Lung Adenocarcinoma," Journal of Cancer Therapy, Vol. 3 No. 4A, 2012, pp. 397-405. doi: 10.4236/jct.2012.324052.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] J. Folkman, E. Merler, C. Abernathy and G. Williams, “Isolation of a Tumor Factor Responsible for Angiogenesis,” Journal of Experimental Medicine, Vol. 133, No. 2, 1971, pp. 275-288. doi:10.1084/jem.133.2.275
[2] P. Wachsberger, R. Burd and A. P. Dicker, “Tumor Response to Ionizing Radiation Combined with Antiangiogenesis or Vascular Targeting Agents: Exploring Mechanisms of Interaction,” Clinical Cancer Research, Vol. 9, No. 6, 2003, pp. 1957-1971.
[3] Y. Cao, R. Cao and N. Veitonmaki, “Kringle Structures and Antiangiogenesis,” Current Medicinal Chemistry— Anti-Cancer Agents, Vol. 2, No. 6, 2002, pp. 667-681. doi:10.2174/1568011023353705
[4] D. J. Davidson, C. Haskell, S. Majest, A. Kherzai, D. A. Egan, K. A. Walter, et al., “Kringle 5 of Human Plasminogen Induces Apoptosis of Endothelial and Tumor Cells through Surface-Expressed Glucose-Regulated Protein 78,” Cancer Research, Vol. 65, No. 11, 2005, pp. 4663-4672. doi:10.1158/0008-5472.CAN-04-3426
[5] L. Bello, C. Giussani, G. Carrabba, M. Pluderi, F. Costa and A. Bikfalvi, “Angiogenesis and Invasion in Gliomas,” Cancer Treatment and Research, Vol. 117, 2004, pp. 263-284. doi:10.1007/978-1-4419-8871-3_16
[6] G.-H. Jin, D.-Y. Ma, N. Wu, F. M. M. T. Marikar, S.-Z. Jin, W.-W. Jiang, et al., “Combination of Human Plasminogen Kringle 5 with Ionizing Radiation Significantly Enhances the Efficacy of Antitumor Effect,” International Journal of Cancer, Vol. 121, No. 11, 2007, pp. 2539-2546. doi:10.1002/ijc.22708
[7] M. M. Ahmed, “Regulation of Radiation-Induced Apoptosis by Early Growth Response-1 Gene in Solid Tumors,” Current Cancer Drug Targets, Vol. 4, No. 1, 2004, pp. 43-52. doi:10.2174/1568009043481704
[8] O. Greco, B. Marples, G. U. Dachs, K. J. Williams, A. V. Patterson and S. D. Scott, “Novel Chimeric Gene Promoters Responsive to Hypoxia and Ionizing Radiation,” Gene Therapy, Vol. 9, No. 20, 2002, pp. 1403-1411. doi:10.1038/
[9] M. Y. Wu, X. Y. Wu, Q. S. Li and R. M. Zheng, “Expression of Egr-1 Gene and Its Correlation with the Oncogene Proteins in Non-Irradiated and Irradiated Esophageal Squamous Cell Carcinoma,” Diseases of the Esophagus, Vol. 19, No. 4, 2006, pp. 267-272. doi:10.1111/j.1442-2050.2006.00575.x
[10] H. B. Ma, X. J. Wang, Z. L. Di, H. Xia, Z. Li, J. Liu, et al., “Construction of Targeted Plasmid Vector pcDNA3.1-Egr.1p-p16 and Its Expression in Pancreatic Cancer JF305 Cells Induced by Radiation in Vitro,” World Journal of Gastroenterology, Vol. 13, No. 31, 2007, pp. 4214-4218.
[11] N. Khalighinejad, H. Hariri, O. Behnamfar, A. Yousefi and A. Momeni, “Adenoviral Gene Therapy in Gastric Cancer: A Review,” World Journal of Gastroenterology, Vol. 14, No. 2, 2008, pp. 180-184. doi:10.3748/wjg.14.180
[12] F. M. Boyce and N. L. Bucher, “Baculovirus-Mediated Gene Transfer into Mammalian Cells,” Proceedings of the National Academy of Sciences USA, Vol. 93, No. 6, 1996, pp. 2348-2352. doi:10.1073/pnas.93.6.2348
[13] H. Matilainen, J. Rinne, L. Gilbert, V. Marjomaki, H. Reunanen and C. Oker-Blom, “Baculovirus Entry into Human Hepatoma Cells,” Journal of Virology, Vol. 79, No. 24, 2005, pp. 15452-15459. doi:10.1128/JVI.79.24.15452-15459.2005
[14] D. L. Jarvis and A. Garcia Jr., “Long-Term Stability of Baculoviruses Stored under Various Conditions,” Biotechniques, Vol. 16, 1994, pp. 508-513.
[15] W. Huang, X. L. Tian, Y. L. Wu, J. Zhong, L. F. Yu, S. P. Hu, et al., “Suppression of Gastric Cancer Growth by Baculovirus Vectormediated Transfer of Normal Epithelial Cell Specific-1 Gene,” World Journal of Gastroenterology, Vol. 14, No. 38, 2008, pp. 5810-5815. doi:10.3748/wjg.14.5810
[16] R. Guo, Y. F. Zhang, S. Liang, H. P. Xu, M. Zhang and B. Li, “Sodium Butyrate Enhances the Expression of Baculovirus-Mediated Sodium/Iodide Symporter Gene in A549 Lung Adenocarcinoma Cells,” Nuclear Medicine Communications, Vol. 31, No. 10, 2010, pp. 916-921.
[17] J. M. McLoughlin, T. M. McCarty, C. Cunningham, V. Clark, N. Senzer, J. Nemunaitis, et al., “TNFerade, an Adenovector Carrying the Transgene forHuman Tumor Necrosis Factor a, for Patients with Advanced Solid Tumors: Surgical Experience and Long-Term Follow-Up,” Annals of Surgical Oncology, Vol.12, No.10, 2005, pp. 825-830. doi:10.1245/ASO.2005.03.023
[18] R. P. Dings, B. W. Williams, C. W. Song, A. W. Griffioen, K. H. Mayo and R. J. Griffin, “Anginex Synergizes with Radiation Therapy to Inhibit Tumor Growth by Radiosensitizing Endothelial Cells,” International Journal of Cancer, Vol. 115, No. 2, 2005, pp. 312-319. doi:10.1002/ijc.20850
[19] S. Nasu, K. K. Ang, Z. Fan and L. Milas, “C225 Antiepidermal Growth Factor Receptor Antibody Enhances Tumor Radiocurability,” International Journal of Radiation Oncology Biology Physics, Vol. 51, No. 2, 2001, pp. 474-477.
[20] J. P. Condreay and T. A. Kost, “Baculovirus Expression Vectors for Insect and Mammalian Cells,” Current Drug Targets, Vol. 8, No. 10, 2007, pp. 1126-1131. doi:10.2174/138945007782151351
[21] C. Hofmann, V. Sandig, G. Jennings, M. Rudolph, P. Schlag and M. Strauss, “Efficient Gene Transfer into Human Hepatocytes by Baculovirus Vectors,” Proceedings of the National Academy of Sciences of the USA, Vol. 92, 1995, pp. 10099-10103. doi:10.1073/pnas.92.22.10099
[22] T. A. Kost and J. P. Condreay, “Recombinant Baculoviruses as Mammalian Cell Gene-Delivery Vectors,” Trends in Biotechnology, Vol. 20, No. 4, 2002, pp. 173-180. doi:10.1016/S0167-7799(01)01911-4
[23] S. Ghosh, M. K. Parvez, K. Banerjee, S. K. Sarin and S. E. Hasnain, “Baculovirus as Mammalian Cell Expression Vector for Gene Therapy: An Emerging Strategy,” Molecular Therapy, Vol. 6, No. 1, 2002, pp. 5-11. doi:10.1006/mthe.2000.0643

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