Establishment of a colorectal cancer nude mouse visualization model of HIF-1α overexpression

Li,Zhe; Wang,Jiye; Zhou,Tianbao; Ye,Xiaolei

doi:10.3892/ol.2016.4287

Establishment of a colorectal cancer nude mouse visualization model of HIF-1α overexpression

Authors:
- Zhe Li
- Jiye Wang
- Tianbao Zhou
- Xiaolei Ye
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Affiliations: Department of Thyroid and Breast Surgery, The Shanghai Tenth People's Hospital, Tong Ji University, Shanghai 200072, P.R. China, Department of Criminal Science and Technology, Zhejiang Police College, Hangzhou, Zhejiang 310053, P.R. China, Hepatobiliary Surgery Centre, The Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China, Division of Drugs and Pharmacology, Ningbo Institute of Medical Sciences, Ningbo, Zhejiang 315020, P.R. China
Published online on: March 1, 2016 https://doi.org/10.3892/ol.2016.4287
Pages: 2725-2732
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to establish a model of tumor cell growth and visualize HIF-1α overexpression in a nude mouse xenograft model of colorectal cancer (CRC). In the study, HIF-1α lentiviral vector and helper plasmid were co-transfected into 293T packaging cells using a liposome method, and the virus was collected following transfection and used to infect CRC SW480, SW620, LoVo and HCT116 cells. Puromycin was used for the selection and large‑scale amplification of the stable HIF‑1α expression of green fluorescent protein (GFP)-positive cells. HIF‑1α‑expressing cells were injected intraperitoneally into a nude mouse xenograft model, and resulting tumor nodules was separated and confirmed using an inverted fluorescence microscope. The results demonstrated that HIF-1α was not expressed in CRC cells in normoxic conditions. When treated with CoCl2, the expression of HIF‑1α could be induced in all the cancer cell lines, except SW480. HIF-1α was highly expressed following infection with lentiviral particles. Stable expression of HIF‑1α promoted migration in the SW480 cells. Following intraperitoneal injection of nude mice with SW480‑HIF‑1α, a significant number of tumor nodules formed in the intestinal wall compared with the controls (P<0.05). The successful construction of the dual expression HIF‑1α and GFP visualization xenograft model provides a good foundation for the screening of HIF-1α-related functions and for investigating the therapeutic potential of drugs that target HIF-1α.

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1	Siegel R, Desantis C and Jemal A: Colorectal cancer statistics, 2014. CA Cancer J Clin. 64:104–117. 2014. View Article : Google Scholar : PubMed/NCBI
2	Nishimoto A, Kugimiya N, Hosoyama T, Enoki T, Li TS and Hamano K: HIF-1α activation under glucose deprivation plays a central role in the acquisition of anti-apoptosis in human colon cancer cells. Int J Oncol. 44:2077–2084. 2014.PubMed/NCBI
3	Cao D, Hou M, Guan YS, Jiang M, Yang Y and Gou HF: Expression of HIF-1α and VEGF in colorectal cancer: Association with clinical outcomes and prognostic implications. BMC Cancer. 9:4322009. View Article : Google Scholar : PubMed/NCBI
4	Kim SE, Shim KN, Jung SA, Yoo K and Lee JH: The clinicopathological significance of tissue levels of hypoxia-inducible factor-1α and vascular endothelial growth factor in gastric cancer. Gut Liver. 3:88–94. 2009. View Article : Google Scholar : PubMed/NCBI
5	Lucarini G, Zizzi A, Belvederesi L, Kyriakidou K, Mazzucchelli R and Biagini G: Increased VEGF165 expression in HCT116 colon cancer cells after transient transfection with a GFP vector encoding HIF-1 gene. J Exp Clin Cancer Res. 26:515–519. 2007.PubMed/NCBI
6	Zhong H, De Marzo AM, Laughner E, Lim M, Hilton DA, Zagzag D, Buechler P, Isaacs WB, Semenza GL and Simons JW: Overexpression of hypoxia-inducible factor 1α in common human cancers and their metastases. Cancer Res. 59:5830–5835. 1999.PubMed/NCBI
7	Hongo K, Tsuno NH, Kawai K, Sasaki K, Kaneko M, Hiyoshi M, Murono K, Tada N, Nirei T, Sunami E, et al: Hypoxia enhances colon cancer migration and invasion through promotion of epithelial-mesenchymal transition. J Surg Res. 182:75–84. 2013. View Article : Google Scholar : PubMed/NCBI
8	Dekervel J, Hompes D, van Malenstein H, Popovic D, Sagaert X, De Moor B, Van Cutsem E, D'Hoore A, Verslype C and van Pelt J: Hypoxia-driven gene expression is an independent prognostic factor in stage II and III colon cancer patients. Clin Cancer Res. 20:2159–2168. 2014. View Article : Google Scholar : PubMed/NCBI
9	Ciafrè SA, Niola F, Giorda E, Farace MG and Caporossi D: CoCl(2)-simulated hypoxia in skeletal muscle cell lines: Role of free radicals in gene up-regulation and induction of apoptosis. Free Radic Res. 41:391–401. 2007. View Article : Google Scholar : PubMed/NCBI
10	Cockrell AS and Kafri T: Gene delivery by lentivirus vectors. Mol Biotechnol. 36:184–204. 2007. View Article : Google Scholar : PubMed/NCBI
11	Ciuffi A: Mechanisms governing lentivirus integration site selection. Curr Gene Ther. 8:419–429. 2008. View Article : Google Scholar : PubMed/NCBI
12	McGarrity GJ, Hoyah G, Winemiller A, Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl G, et al: Patient monitoring and follow-up in lentiviral clinical trials. J Gene Med. 15:78–82. 2013. View Article : Google Scholar : PubMed/NCBI
13	Kubis HP, Hanke N, Scheibe RJ and Gros G: Accumulation and nuclear import of HIF1 alpha during high and low oxygen concentration in skeletal muscle cells in primary culture. Biochim Biophys Acta. 1745:187–195. 2005. View Article : Google Scholar : PubMed/NCBI
14	Zhang B, Guo W, Yu L, Wang F, Xu Y, Liu Y and Huang C: Cobalt chloride inhibits tumor formation in osteosarcoma cells through upregulation of HIF-1α. Oncol Lett. 5:911–916. 2013.PubMed/NCBI
15	Wang V, Davis DA, Haque M, Huang LE and Yarchoan R: Differential gene up-regulation by hypoxia-inducible factor-1 alpha and hypoxia-inducible factor-2 alpha in HEK293T cells. Cancer Res. 65:3299–3306. 2005.PubMed/NCBI
16	Onnis B, Rapisarda A and Melillo G: Development of HIF-1 inhibitors for cancer therapy. J Cell Mol Med. 13:2780–2786. 2009. View Article : Google Scholar : PubMed/NCBI
17	Talks KL, Turley H, Gatter KC, Maxwell PH, Pugh CW, Ratcliffe PJ and Harris AL: The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages. Am J Pathol. 157:411–421. 2000. View Article : Google Scholar : PubMed/NCBI
18	Buffa FM, West C, Byrne K, Moore JV and Nahum AE: Radiation response and cure rate of human colon adenocarcinoma spheroids of different size: The significance of hypoxia on tumor control modelling. Int J Radiat Oncol Biol Phys. 49:1109–1118. 2001. View Article : Google Scholar : PubMed/NCBI
19	Ryan HE, Lo J and Johnson RS: HIF-1 alpha is required for solid tumor formation and embryonic vascularization. EMBO J. 17:3005–3015. 1998. View Article : Google Scholar : PubMed/NCBI
20	Tsai YP and Wu KJ: Hypoxia-regulated target genes implicated in tumor metastasis. J Biomed Sci. 19:1022012. View Article : Google Scholar : PubMed/NCBI
21	Li ZH, Liao W, Cui XL, Zhao Q, Liu M, Chen YH, Liu TS, Liu NL, Wang F, Yi Y, et al: Intravenous transplantation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci. 8:74–83. 2011. View Article : Google Scholar : PubMed/NCBI
22	Choy G, O'Connor S, Diehn FE, Costouros N, Alexander HR, Choyke P and Libutti SK: Comparison of noninvasive fluorescent and bioluminescent small animal optical imaging. Biotechniques. 35:1022–1026, 1028-1030. 2003.PubMed/NCBI
23	Ralph GS, Parham S, Lee SR, Beard GL, Craigon MH, Ward N, White JR, Barber RD, Rayner W, Kingsman SM, et al: Identification of potential stroke targets by lentiviral vector mediated overexpression of HIF-1 alpha and HIF-2 alpha in a primary neuronal model of hypoxia. J Cereb Blood Flow Metab. 24:245–258. 2004. View Article : Google Scholar : PubMed/NCBI