Efficacy of pEgr‑1‑endostatin combined with ionizing radiation on hypoxic conditions in nude mice bearing SKOV3 ovarian carcinoma

Zhang,Yong‑Chun; Li,Xiu‑Mei; Yu,Zhuang; Shi,Xiao‑Li; Li,Yong; Wang,Wan‑Lin

doi:10.3892/ol.2017.5559

Efficacy of pEgr‑1‑endostatin combined with ionizing radiation on hypoxic conditions in nude mice bearing SKOV3 ovarian carcinoma

Authors:
- Yong‑Chun Zhang
- Xiu‑Mei Li
- Zhuang Yu
- Xiao‑Li Shi
- Yong Li
- Wan‑Lin Wang
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Affiliations: Department of Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China, Department of Ultrasound, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China, Department of Gynecology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China
Published online on: January 2, 2017 https://doi.org/10.3892/ol.2017.5559
Pages: 1101-1108
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypoxia occurs in a wide range of solid tumors, and is strongly associated with radio‑resistance of malignant tumors. The aim of the present study was to investigate the effect of endostatin combined with ionizing radiation (IR) on hypoxic conditions. A total of 24 mice bearing SKOV3 ovarian carcinoma were divided into three groups. Following injection with pEgr‑1‑endostatin plasmid for 12 h, the mice in the endostatin‑IR‑treated group were exposed to 300 cGy/min X‑ray for 48 h, and the IR‑treated group was exposed to the same condition. Then, the expression of endostatin, hypoxia‑inducible factor (HIF)‑1α and vascular endothelial growth factor (VEGF) was detected by reverse transcription‑polymerase chain reaction, ELISA, immunohistochemistry and western blotting. In addition, the tumor microvessel density (MVD) was examined by immunohistochemistry analysis of cluster of differentiation 31‑positive cells. The results revealed that pEgr‑1‑endostatin was successfully induced by IR. The level of endostatin messenger RNA in the endostatin‑IR‑treated group was significantly higher than that in the control and IR‑treated groups (F=380.078, P<0.001). Statistical differences were also examined at the protein level by western blotting and ELISA. An obvious increase in MVD was observed in the IR‑treated group compared with that in the control group (t=7.040, P<0.001), and a significant decrease in MVD was observed in the endostatin‑IR‑treated group compared with that in the control group (t=18.153, P<0.001). By comparing the morphology of the tumor vasculature in the three groups, it was noticed that the microvessels in the endostatin‑IR‑treated group were more regularly distributed and had fewer giant branches than those in the IR‑treated group. Further investigation revealed that the expression levels of HIF‑1α and VEGF in the endostatin‑IR‑treated group were lower compared with those in the control (t=5.339, P=0.001; and t=13.880, P<0.001, respectively) and the IR‑treated groups (t=12.930, P<0.001; and t=14.050, P<0.001, respectively). Our findings suggested that endostatin decreased the number of microvessels via the HIF‑1/VEGF signaling pathway, and that pEgr‑1‑endostatin combined with IR may improve hypoxic conditions and may be a novel approach for treating solid tumors.

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1	Gray LH, Conger AD, Ebert M, Hornsey S and Scott OC: The concentration of oxygen dissolved in tissues at the time of irradiation as a factor in radiotherapy. Br J Radiol. 26:638–648. 1953. View Article : Google Scholar : PubMed/NCBI
2	Moulder JE and Rockwell S: Tumor hypoxia: Its impact on cancer therapy. Cancer Metastasis Rev. 5:313–341. 1987. View Article : Google Scholar : PubMed/NCBI
3	Holzer LA, Cör A, Pfandlsteiner G and Holzer G: Expression of VEGF, its receptors and HIF-1α in Dupuytren's disease. Acta Orthop. 84:420–425. 2013. View Article : Google Scholar : PubMed/NCBI
4	Brahimi-Horn MC, Chiche J and Pouyssegur J: Hypoxia and cancer. J Mol Med (Berl). 85:1301–1307. 2007. View Article : Google Scholar : PubMed/NCBI
5	Vaupel P, Hockel M and Mayer A: Detection and characterization of tumor hypoxia using pO2 histography. Antioxid Redox Signal. 9:1221–1235. 2007. View Article : Google Scholar : PubMed/NCBI
6	Multhoff G, Radons J and Vaupel P: Critical role of aberrant angiogenesis in the development of tumor hypoxia and associated radioresistance. Cancers (Basel). 6:813–828. 2014. View Article : Google Scholar : PubMed/NCBI
7	Ikeda E: Cellular response to tissue hypoxia and its involvement in disease progression. Pathol Int. 55:603–610. 2005. View Article : Google Scholar : PubMed/NCBI
8	Salceda S and Caro J: Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. J Biol Chem. 272:22642–22647. 1997. View Article : Google Scholar : PubMed/NCBI
9	Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3:721–732. 2003. View Article : Google Scholar : PubMed/NCBI
10	Arjamaa O, Nikinmaa M, Salminen A and Kaarniranta K: Regulatory role of HIF-1alpha in the pathogenesis of age-related macular degeneration (AMD). Ageing Res Rev. 8:349–358. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zhang P, Zhang X, Hao X, Wang Y, Hui Y, Wang H, Hu D and Zhou J: Rac1 activates HIF-1 in retinal pigment epithelium cells under hypoxia. Graefes Arch Clin Exp Ophthalmol. 247:633–639. 2009. View Article : Google Scholar : PubMed/NCBI
12	Semenza GL: Regulation of mammalian O2 homeostasis by hypoxia-inducible factor 1. Annu Rev Cell Dev Biol. 15:551–578. 1999. View Article : Google Scholar : PubMed/NCBI
13	Tammela T, Zarkada G, Nurmi H, Jakobsson L, Heinolainen K, Tvorogov D, Zheng W, Franco CA, Murtomäki A, Aranda E, et al: VEGFR-3 controls tip to stalk conversion at vessel fusion sites by reinforcing Notch signalling. Nat Cell Biol. 13:1202–1213. 2011. View Article : Google Scholar : PubMed/NCBI
14	Semenza GL: HIF-1 and human disease: One highly involved factor. Genes Dev. 14:1983–1991. 2000.PubMed/NCBI
15	Jain RK: Normalization of tumor vasculature: An emerging concept in antiangiogenic therapy. Science. 307:58–62. 2005. View Article : Google Scholar : PubMed/NCBI
16	Ke QH, Zhou SQ, Huang M, Lei Y, Du W and Yang JY: Early efficacy of Endostar combined with chemoradiotherapy for advanced cervical cancers. Asian Pac J Cancer Prev. 13:923–926. 2012. View Article : Google Scholar : PubMed/NCBI
17	O'Reilly MS, Boehm T, Shing Y, Fukai N, Vasios G, Lane WS, Flynn E, Birkhead JR, Olsen BR and Folkman J: Endostatin: An endogenous inhibitor of angiogenesis and tumor growth. Cell. 88:277–285. 1997. View Article : Google Scholar : PubMed/NCBI
18	Qi X, Liu Y, Wei W, Huang X and Zuo Y: Effects of the C-terminal of endostatin on the tumorigenic potential of H22 cells. Biomed Rep. 1:761–765. 2013.PubMed/NCBI
19	Huang RP and Adamson ED: A biological role for Egr-1 in cell survival following ultra-violet irradiation. Oncogene. 10:467–475. 1995.PubMed/NCBI
20	Zhang Y, Qiu W, Liang J, Yu Z and Yue L: Anti-tumor effects of pEgr-1-endostatin-TNF-α recombinant plasmid expression induced by ionizing radiation. Asian Pac J Cancer Prev. 12:2933–2937. 2011.PubMed/NCBI
21	El-Gendi S and Abdel-Hadi M: Lymphatic vessel density as prognostic factor in breast carcinoma: Relation to clinicopathologic parameters. J Egypt Natl Canc Inst. 21:139–149. 2009.PubMed/NCBI
22	Ferrara N and Davis-Smyth T: The biology of vascular endothelial growth factor. Endocr Rev. 18:4–25. 1997. View Article : Google Scholar : PubMed/NCBI
23	Cao D, Hou M, Guan YS, Jiang M, Yang Y and Gou HF: Expression of HIF-1alpha and VEGF in colorectal cancer: Association with clinical outcomes and prognostic implications. BMC Cancer. 9:4322009. View Article : Google Scholar : PubMed/NCBI
24	Wilson WR and Hay MP: Targeting hypoxia in cancer therapy. Nat Rev Cancer. 11:393–410. 2011. View Article : Google Scholar : PubMed/NCBI
25	Li L, Huang JL, Liu QC, Wu PH, Liu RY, Zeng YX and Huang WL: Endostatin gene therapy for liver cancer by a recombinant adenovirus delivery. World J Gastroenterol. 10:1867–1871. 2004. View Article : Google Scholar : PubMed/NCBI
26	Yoshimura M, Itasaka S, Harada H and Hiraoka M: Microenvironment and radiation therapy. Biomed Res Int. 2013:6853082013. View Article : Google Scholar : PubMed/NCBI
27	Wang J, Sun Y, Liu Y, Yu Q, Zhang Y, Li K, Zhu Y, Zhou Q, Hou M, Guan Z, et al: Results of randomized, multicenter, double-blind phase III trial of rh-endostatin (YH-16) in treatment of advanced non-small cell lung cancer patients. Zhongguo Fei Ai Za Zhi. 8:283–290. 2005.(In Chinese). PubMed/NCBI
28	Folkman J: Tumor angiogenesis: Therapeutic implications. N Engl J Med. 285:1182–1186. 1971. View Article : Google Scholar : PubMed/NCBI
29	Forsythe JA, Jiang BH, Iyer NV, Agani F, Leung SW, Koos RD and Semenza GL: Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol. 16:4604–4613. 1996. View Article : Google Scholar : PubMed/NCBI
30	Brizel DM, Scully SP, Harrelson JM, Layfield LJ, Bean JM, Prosnitz LR and Dewhirst MW: Tumor oxygenation predicts for the likelihood of distant metastases in human soft tissue sarcoma. Cancer Res. 56:941–943. 1996.PubMed/NCBI
31	Lohela M, Bry M, Tammela T and Alitalo K: VEGFs and receptors involved in angiogenesis versus lymphangiogenesis. Curr Opin Cell Biol. 21:154–165. 2009. View Article : Google Scholar : PubMed/NCBI
32	Mauceri HJ, Beckett MA, Liang H, Sutton HG, Pitroda S, Galka E, Efimova E, Darga T, Khodarev NN, King CR, et al: Translational strategies exploiting TNF-alpha that sensitize tumors to radiation therapy. Cancer Gene Ther. 16:373–381. 2009. View Article : Google Scholar : PubMed/NCBI
33	Kim YM, Jang JW, Lee OH, Yeon J, Choi EY, Kim KW, Lee ST and Kwon YG: Endostatin inhibits endothelial and tumor cellular invasion by blocking the activation and catalytic activity of matrix metalloproteinase. Cancer Res. 60:5410–5413. 2000.PubMed/NCBI
34	Rehn M, Veikkola T, Kukk-Valdre E, Nakamura H, Ilmonen M, Lombardo C, Pihlajaniemi T, Alitalo K and Vuori K: Interaction of endostatin with integrins implicated in angiogenesis. Proc Natl Acad Sci USA. 98:1024–1029. 2001. View Article : Google Scholar : PubMed/NCBI