Radiation-inducible promoter mediated PUMA Gene in the the treatment of Tca8113

Dong-Sheng Yu, Wei Zhao, Jia-xian Luo, Bing Liu, Xiao-Lin Wu
DOI: 10.4236/ojst.2011.14030                

Abstract

Objective: The aim of this study is to explore the therapeutic effect of the PUMA gene mediated by ra- diation-inducible promoters in the treatment of ton- gue squamous cell carcinoma. Methods: Recombi- nant pcDNA3.1 (+)/E-PUMA was constructed, in which the PUMA gene was mediated by a synthetic radiation inducible promoter. The recombined plas-mids were transfected into the Tca8113 cell and xeno- grafts of human tongue squamous carcinoma in na- ked mice respectively. After 24 h, the tumors were tr- eated with 3 Gy of irradiation to upregulate the PU- MA gene expression. PUMA mRNA was detected by RT-PCR. Proliferating cell nuclear antigen (PCNA) and apoptosis were detected by immunohistochemi- cal method and in situ end-labeling (ISEL) respecti- vely. The data were analyzed using the SPSS11.0 soft- ware package for chi-square test. RESULTS: Com-pared with the control group, the comparative survi- val rate of Tca8113 cells in the PUMA/IR group was markedly decreased and the xenografts were signify- cantly suppressed. Up-regulation of PUMA gene ex- pression was observed in the Tca8113 cells and in the xenografts after irradiation. The apoptosis indices of the Tca8113 cells and xenograft with irradiation were markedly higher than those without irradiation. At the same time, the proliferation indices of the Tca- 8113 cells and xenografts with irradiation were mark- edly lower than those without irradiation. Conclu- sions: radiation-induc ible promoters can serve as molecular switches to improve the expression of PU- MA gene in tongue squamous cell carcinoma both in vivo and in vitro. Low-dose induction radiation can significantly improve therapeutic efficiency.

Keywords

Blast; Graph Theory; Redundant Sequences; CD-HIT

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Nakano, K. and Vousden, K.H. (2001) PUMA, a novel proapoptotic gene, is induced by p53. Molecular Cell, 7, 683-694. doi:10.1016/S1097-2765(01)00214-3
[2]	Labi, V. and Villunger, A. (2010) PUMA-mediated tumor suppression: A tale of two stories. Cell Cycle, 9, 4269-4275. doi:10.4161/cc.9.21.13666
[3]	Avila, J.L., Grundmann, O., Burd, R., et al. (2009) Radiation-induced salivary gland dysfunction results from p53-dependent apoptosis. International Journal of Radiation Oncology, Biology, Physics, 2, 523-529. doi:10.1016/j.ijrobp.2008.09.036
[4]	Chen, Y., Xiang, Y. and Lin, C. (2007) PUMA—A Promising tumor gene therapy target. Oncology Progress, 5, 282-285.
[5]	Michalak, E.M., Vandenberg, C.J., Delbridg, A.R., et al., (2010) Apoptosis-promoted tumorigenesis: Gamma- irradiation-induced thymic lymph omagenesis requires. Puma-driven leukocyte death Genes Dev, 24, 1608-1613. doi:10.1101/gad.1940110
[6]	Shao, L., Sun, Y., Zhang, Z., et al. (2010) Deletion of proapoptotic PUMA selectively protects hematopo- ietic stem and progenitor cells against high-dose radiation. Blood, 23, 4707-4714. doi:10.1182/blood-2009-10-248872
[7]	Yu, D.-S., Zhao, W., Huang, H.-Z., et al. (2010) Syn- thetic radiation-inducible promoters mediated HSV- TK/GCV gene therapy in the treatment of oral squamous cell carcinoma. Oral Diseases, 16, 445-452. doi:10.1111/j.1601-0825.2010.01655.x
[8]	Rao, G.V., Kumar, N.A., Gupta, R., et al. (2010) Radiation-inducible silencing of uPA and uPAR in vitro and in vivo in meningioma. International Journal of Oncology, 4, 809-816.
[9]	Nandi, S., Ulasov, I.V., Tyler, M.A., et al. (2008) Low-dose radiation enhances surviving-mediated virotherapy against malignant glioma stem cells. Cancer Research, 14, 5778-5784. doi:10.1158/0008-5472.CAN-07-6441
[10]	Soudry, E., Preis, M., Hod, R., et al. (2010) Squamous cell carcinoma of the oral tongue in patients younger than 30 years: Clinicopathologic features and outcome. Clinical Otolaryngology, 4, 307-312. doi:10.1111/j.1749-4486.2010.02164.x
[11]	Zhang, T.H., Liu, H.C., Zhu, L.J., et al. (2011) Activation of notch signaling in human tongue carcinoma. Journal of Oral Pathology & Medicine, 1, 37-45. doi:10.1111/j.1600-0714.2010.00931.x
[12]	Elias, S.T., Diniz, J., Almeida, R.S., et al. (2010) Cytotoxic effect of tobacco extracts on human oral squamous cell carcinoma cell-line. Oral Oncology, 12, 869-873. doi:10.1016/j.oraloncology.2010.09.008
[13]	Hajji, N. and Joseph, B. (2010) Epigenetic regulation of cell life and death decisions and deregulation in cancer. Eessays in Biochemistry, 1, 121-146. doi:10.1042/bse0480121
[14]	Kerr, J.F., Wyllie, A.H. and Currie, A.R. (1972) Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. British Journal of Cancer, 4, 239-257. doi:10.1038/bjc.1972.33
[15]	Qiu, W., Leibowitz, B., Zhang, L., et al. (2010) Growth factors protect intestinal stem cells from radiation-induced apoptosis by suppressing PUMA through the PI3K/AKT/p53 axis. Oncogene, 11, 1622- 1632. doi:10.1038/onc.2009.451
[16]	Chipuk, J.E. and Green, D.R. (2009) PUMA cooperates with direct activator proteins to promote mitochondrial outer membrane permeabilization and apoptosis. Cell Cycle, 17, 2692-2696. doi:10.4161/cc.8.17.9412
[17]	Yu, J. and Zhang, L. (2008) PUMA, a potent killer with or without p53. Oncogene, 27, S71-S83. doi:10.1038/onc.2009.45
[18]	Hong, L.Z., Zhao, X.Y. and Zhang, H.L. (2010) p53-mediated neuronal cell death in ischemic brain injury. Neuroscience Bulletin, 3, 232-240. doi:10.1007/s12264-010-1111-0
[19]	Shaltouki, A., Freer, M., Mei, Y., et al. (2007) Increased expression of the pro-apoptotic Bcl2 family member PUMA is required for mitochondrial release of cytochrome C and the apoptosis associated with skeletal myoblast differentiation. Apoptosis, 12, 2143- 2154. doi:10.1007/s10495-007-0135-z
[20]	Wang, H., Qian, H., Yu, J., et al. (2006) Administration of PUMAadenovirus increases the sensitivity of esophageal cancer cellsto anticancer drugs. Cancer Biology & Therapy, 4, 380-389. doi:10.4161/cbt.5.4.2477
[21]	Hideaki, I., Takao, K. and Toru, M. (2005) Efficacy of PUMA for malignant glioma cells regard less of the p53 status. Human Gene Therapy, 6, 685-692.
[22]	Vousden, K.H. (2005) Apotosis. p53 and PUMA: A deadly duo. Science, 5741, 1685-1686. doi:10.1126/science.1118232
[23]	Yu, J., Yue, W., Wu, B., et al. (2006) PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation. Clinical Cancer Research, 9, 2928-2935. doi:10.1158/1078-0432.CCR-05-2429
[24]	Qiu, W., Carson-Walter, E.B., Liu, H., et al. (2008) PUMA regulates intestinal progenitor cell radio-sen- sitivity and gastrointestinal syndrome. Cell Stem Cell, 6, 576-583. doi:10.1016/j.stem.2008.03.009
[25]	Sasano, N., Enomoto, A., Hosoi, Y., et al. (2010) Eda- ravone, a known free radical scavenger, enhances X-ray-induced apoptosis at low concentrations. Cancer Letters, 1, 52-57. doi:10.1016/j.canlet.2009.12.020
[26]	Tsurushima, H., Yuan, X., Dillehay, L.E., et al. (2008) Radiation-inducible caspase-8 gene therapy for malignant brain tumors. International Journal of Radiation Oncology, Biology, Physics, 2, 517-525. doi:10.1016/j.ijrobp.2008.02.002
[27]	Marignol, L., Coffey, M., Hollywood, D., et al. (2007) Radiation to control transgene expression in tumors. Cancer Biology & Therapy, 7, 1005-1012. doi:10.4161/cbt.6.7.4477
[28]	Vilaboa, N. and Voellmy, R. (2006) Regulatable gene expression systems for gene therapy. Current Gene Therapy, 4, 421-38. doi:10.2174/156652306777934829
[29]	Bhide, S.A. and Nutting, C.M. (2010) Recent advances in radiotherapy. BMC Medicine, 8, 25-28. doi:10.1186/1741-7015-8-25
[30]	Brahme, A. and Lind, B.K. (2010) A systems biology approach to radiation therapy optimization. Radiation and Environmental Biophysics, 2, 111-124. doi:10.1007/s00411-010-0268-2
[31]	John-Aryankalayil, M., Palayoor, S.T., Cerna, D., et al. (2010) Fractionated radiation therapy can induce a molecular profile for therapeutic targeting. Radiation Research, 4, 446-458. doi:10.1667/RR2105.1
[32]	Marignol, L., Robson, T., McCarthy, H.O., et al. (2008) The tissue plasminogen activator gene promoter: A novel tool for radiogenic gene therapy of the prostate? Journal of Gene Medicine, 9, 1032-1038. doi:10.1002/jgm.1221