Synergistic cytotoxic effects of recombinant human adenovirus p53 and radiation at various time points in A549 lung adenocarcinoma cells

Ma,Jie-Tao; Han,Cheng-Bo; Zhao,Jian-Zhu; Jing,Wei; Zhou,Yang; Huang,Le-Tian; Zou,Hua-Wei

doi:10.3892/ol.2012.747

September 2012 Volume 4 Issue 3

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September 2012 Volume 4 Issue 3

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Article

Synergistic cytotoxic effects of recombinant human adenovirus p53 and radiation at various time points in A549 lung adenocarcinoma cells

Authors:
- Jie-Tao Ma
- Cheng-Bo Han
- Jian-Zhu Zhao
- Wei Jing
- Yang Zhou
- Le-Tian Huang
- Hua-Wei Zou
View Affiliations / Copyright

Affiliations: Department of Oncology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110022, P.R. China
Pages: 529-533
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Published online on: June 8, 2012

https://doi.org/10.3892/ol.2012.747
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Abstract

The aim of this study was to evaluate the effects of recombinant human adenovirus p53 (rAd‑p53; Gendicine) transfection and radiation at various time points following transfection. Cytotoxic effects and p53 protein expression levels were analyzed. rAd‑p53 containing the human wild‑type p53 gene was introduced into the human lung adenocarcinoma cell line A549, and cells were irradiated with a single dose of 6 MeV 4 Gy β rays. According to the time interval between rAd-p53 transfection and radiotherapy (RT), A549‑transfected rAd‑p53 cells were divided into 5 groups: radiation administered immediately after transfection (0 h‑RT) group, after 3 h group (3 h‑RT), after 6 h group (6 h‑RT), after 24 h group (24 h‑RT) and after 48 h group (48 h‑RT). Cells with rAd‑p53 transfection alone (Ad-p53) and with empty adenovirus (Ad) were included as the two control groups. Following 72 h of transfection, cell viability and growth were analyzed using MTT assays and flow cytometry, and p53 protein expression was analyzed using western blot analysis. From 0 h‑RT to 48 h‑RT, cell viability gradually decreased, while percentage of apoptotic cells and p53 protein expression gradually increased. The cell viability suppression rates in the 6 h‑RT, 24 h‑RT and 48 h‑RT groups were 56.7±5.4, 60.8±6.0 and 68.9±6.6, respectively, which were significantly greater compared to that of the Ad-p53 (40.8±4.7), 0 h-RT (45.0±3.5) and 3 h-RT groups (47.0±4.3). No statistically significant differences were observed in the cell viability suppression rates among the 6 h-RT, 24 h-RT and 48 h-RT groups (P>0.05). Similar changes were observed in the percentage of apoptotic cells. The p53 protein expression level in the 6 h-RT group (0.856±0.092) was higher compared to that in the 3 h-RT group (0.643±0.089) (t=2.882; P=0.045), but not significantly different from that of the 24 h-RT group (1.193±0.202). The cell viability suppression rate and percentage of apoptotic cells was positively correlated with p53 protein expression in the A549 cells (P<0.05). Radiation may inhibit or damage p53 protein expression at the early stage of rAd-p53 transfection. To sensitize tumor cells to irradiation and achieve maximal cytotoxic effects, it is recommended to conduct RT at least 6 h following transfection with rAd-p53.

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1	Vousden KH and Lu X: Live or let die: the cell's response to p53. Nat Rev Cancer. 2:594–604. 2002. View Article : Google Scholar
2	Hirao A, Kong YY, Matsuoka S, Wakeham A, Ruland J, Yoshida H, Liu D, Elledge SJ and Mak TW: DNA damage-induced activation of p53 by the checkpoint kinase Chk2. Science. 287:1824–1827. 2000. View Article : Google Scholar : PubMed/NCBI
3	Huang CL, Yokomise H and Miyatake A: Clinical significance of the p53 pathway and associated gene therapy in non-small cell lung cancers. Future Oncol. 3:83–93. 2007. View Article : Google Scholar : PubMed/NCBI
4	Kandioler D, Stamatis G, Eberhardt W, Kappel S, Zöchbauer-Müller S, Kührer I, Mittlböck M, Zwrtek R, Aigner C, Bichler C, et al: Growing clinical evidence for the interaction of the p53 genotype and response to induction chemotherapy in advanced non-small cell lung cancer. J Thorac Cardiovasc Surg. 135:1036–1041. 2008. View Article : Google Scholar : PubMed/NCBI
5	Kandioler D, Zwrtek R, Ludwig C, Janschek E, Ploner M, Hofbauer F, Kührer I, Kappel S, Wrba F, Horvath M, et al: TP53 genotype but not p53 immunohistochemical result predicts response to preoperative short-term radiotherapy in rectal cancer. Ann Surg. 235:493–498. 2002. View Article : Google Scholar : PubMed/NCBI
6	Shiomitsu K, Sajo E, Xia X, Hunley DW, Mauldin GE, Li S and Mauldin GN: Radiosensitivity of canine osteosarcoma cells transfected with wild-type p53 in vitro. Vet Comp Oncol. 6:193–200. 2008. View Article : Google Scholar : PubMed/NCBI
7	Gudkov AV and Komarova EA: The role of p53 in determining sensitivity to radiotherapy. Nat Rev Cancer. 3:117–129. 2003. View Article : Google Scholar : PubMed/NCBI
8	Cuddihy AR and Bristow RG: The p53 protein family and radiation sensitivity: yes or no? Cancer Metastasis Rev. 23:237–257. 2004. View Article : Google Scholar : PubMed/NCBI
9	Takahashi T, Fukawa T, Hirayama R, Yoshida Y, Musha A, Furusawa Y, Ando K and Nakano T: In vitro interaction of high-LET heavy-ion irradiation and chemotherapeutic agents in two cell lines with different radiosensitivities and different p53 status. Anticancer Res. 30:1961–1967. 2010.PubMed/NCBI
10	Takahashi A, Matsumoto H, Yuki K, Yasumoto J, Kajiwara A, Aoki M, Furusawa Y, Ohnishi K and Ohnishi T: High-LET radiation enhanced apoptosis but not necrosis regardless of p53 status. Int J Radiat Oncol Biol Phys. 60:591–597. 2004. View Article : Google Scholar : PubMed/NCBI
11	Pan JJ, Zhang SW, Chen CB, Xiao SW, Sun Y, Liu CQ, Su X, Li DM, Xu G, Xu B and Lu YY: Effect of recombinant adenovirus-p53 combined with radiotherapy on long-term prognosis of advanced nasopharyngeal carcinoma. J Clin Oncol. 27:799–804. 2009. View Article : Google Scholar : PubMed/NCBI
12	Roth JA: Adenovirus p53 gene therapy. Expert Opin Biol Ther. 6:55–61. 2006. View Article : Google Scholar : PubMed/NCBI
13	Ma JT, Zheng W and Zou HW: Radiosensitivity enhanced by recombinant adenovirus p53 in lung adenocarcinoma. Journal of China Medical University. 36:424–426. 2007.
14	Ma G, Kawamura K, Li Q, Suzuki N, Liang M, Namba M, Shimada H and Tagawa M: Cytotoxicity of adenoviruses expressing the wild-type p53 gene to esophageal carcinoma cells is linked with the CAR expression level and indirectly with the endogenous p53 status. Cancer Gene Ther. 16:832–840. 2009. View Article : Google Scholar : PubMed/NCBI
15	Ashcroft M, Kubbutat MH and Vousden KH: Regulation of p53 function and stability by phosphorylation. Mol Cell Biol. 19:1751–1758. 1999.PubMed/NCBI
16	Reed M, Woelker B, Wang P, Wang Y, Anderson ME and Tegtmeyer P: The C-terminal domain of p53 recognizes DNA damaged by ionizing radiation. Proc Natl Acad Sci USA. 92:9455–9459. 1995. View Article : Google Scholar : PubMed/NCBI
17	Lakin ND and Jackson SP: Regulation of p53 in response to DNA damage. Oncogene. 18:7644–7655. 1999. View Article : Google Scholar : PubMed/NCBI
18	Gudkov AV and Komarova EA: Prospective therapeutic applications of p53 inhibitors. Biochem Biophys Res Commun. 331:726–736. 2005. 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

Synergistic cytotoxic effects of recombinant human adenovirus p53 and radiation at various time points in A549 lung adenocarcinoma cells

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