Hypoxia-targeted triple suicide gene therapy radiosensitizes human colorectal cancer cells

Hsiao,Hung  Tsung; Xing,Ligang; Deng,Xuelong; Sun,Xiaorong; Ling,C.  Clifton; Li,Gloria  C.

doi:10.3892/or.2014.3238

Hypoxia-targeted triple suicide gene therapy radiosensitizes human colorectal cancer cells

Authors:
- Hung Tsung Hsiao
- Ligang Xing
- Xuelong Deng
- Xiaorong Sun
- C. Clifton Ling
- Gloria C. Li
View Affiliations

Affiliations: Department of Radiation Oncology, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA, Department of Medical Physics, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA
Published online on: June 6, 2014 https://doi.org/10.3892/or.2014.3238
Pages: 723-729

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The hypoxic microenvironment, an important feature of human solid tumors but absent in normal tissue, may provide an opportunity for cancer-specific gene therapy. The purpose of the present study was to investigate whether hypoxia-driven triple suicide gene TK/CD/UPRT expression enhances cytotoxicity to ganciclovir (GCV) and 5-fluorocytosine (5-FC), and sensitizes human colorectal cancer to radiation in vitro and in vivo. Stable transfectant of human colorectal HCT8 cells was established which expressed hypoxia-inducible vectors (HRE-TK/eGFP and HRE-CD/UPRT/mDsRed). Hypoxia-induced expression/function of TK, CD and UPRT was verified by western blot analysis, flow cytometry, fluorescent microscopy and cytotoxicity assay of GCV and 5-FC. Significant radiosensitization effects were detected after 5-FC and GCV treatments under hypoxic conditions. In the tumor xenografts, the distribution of TK/eGFP and CD/UPRT/mDsRed expression visualized with fluorescence microscopy was co-localized with the hypoxia marker pimonidazole positive staining cells. Furthermore, administration of 5-FC and GCV in mice in combination with local irradiation resulted in tumor regression, as compared with prodrug or radiation treatments alone. Our data suggest that the hypoxia-inducible TK/GCV+CDUPRT/5-FC triple suicide gene therapy may have the ability to specifically target hypoxic cancer cells and significantly improve the tumor control in combination with radiotherapy.

View Figures

View References

1	Wilson WR and Hay MP: Targeting hypoxia in cancer therapy. Nat Rev Cancer. 11:393–410. 2011. View Article : Google Scholar
2	Nordsmark M, Bentzen SM, Rudat V, et al: Prognostic value of tumor oxygenation in 397 head and neck tumors after primary radiation therapy. An international multi-center study. Radiother Oncol. 77:18–24. 2005. View Article : Google Scholar : PubMed/NCBI
3	Brizel DM, Scully SP, Harrelson JM, et al: Tumor oxygenation predicts for the likelihood of distant metastases in human soft tissue sarcoma. Cancer Res. 56:941–943. 1996.PubMed/NCBI
4	Rampling R, Cruickshank G, Lewis AD, Fitzsimmons SA and Workman P: Direct measurement of pO₂ distribution and bioreductive enzymes in human malignant brain tumors. Int J Radiat Oncol Biol Phys. 29:427–431. 1994.PubMed/NCBI
5	Keith B, Johnson RS and Simon MC: HIF1α and HIF2α: sibling rivalry in hypoxic tumour growth and progression. Nat Rev Cancer. 12:9–22. 2012.
6	Wang GL and Semenza GL: General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia. Proc Natl Acad Sci USA. 90:4304–4308. 1993. View Article : Google Scholar : PubMed/NCBI
7	Marignol L, Lawler M, Coffey M and Hollywood D: Achieving hypoxia-inducible gene expression in tumors. Cancer Biol Ther. 4:359–364. 2005. View Article : Google Scholar : PubMed/NCBI
8	Greco O, Marples B, Joiner MC and Scott SD: How to overcome (and exploit) tumor hypoxia for targeted gene therapy. J Cell Physiol. 197:312–325. 2003. View Article : Google Scholar : PubMed/NCBI
9	Marignol L, Foley R, Southgate TD, Coffey M, Hollywood D and Lawler M: Hypoxia response element-driven cytosine deaminase/5-fluorocytosine gene therapy system: a highly effective approach to overcome the dynamics of tumour hypoxia and enhance the radiosensitivity of prostate cancer cells in vitro. J Gene Med. 11:169–179. 2009. View Article : Google Scholar
10	Koshikawa N, Takenaga K, Tagawa M and Sakiyama S: Therapeutic efficacy of the suicide gene driven by the promoter of vascular endothelial growth factor gene against hypoxic tumor cells. Cancer Res. 60:2936–2941. 2000.PubMed/NCBI
11	Freytag SO, Stricker H, Pegg J, et al: Phase I study of replication-competent adenovirus-mediated double-suicide gene therapy in combination with conventional-dose three-dimensional conformal radiation therapy for the treatment of newly diagnosed, intermediate- to high-risk prostate cancer. Cancer Res. 63:7497–7506. 2003.
12	Rogulski KR, Zhang K, Kolozsvary A, Kim JH and Freytag SO: Pronounced antitumor effects and tumor radiosensitization of double suicide gene therapy. Clin Cancer Res. 3:2081–2088. 1997.PubMed/NCBI
13	Xing L, Sun X, Deng X, et al: Expression of the bifunctional suicide gene CDUPRT increases radiosensitization and bystander effect of 5-FC in prostate cancer cells. Radiother Oncol. 92:345–352. 2009. View Article : Google Scholar : PubMed/NCBI
14	Porosnicu M, Mian A and Barber GN: The oncolytic effect of recombinant vesicular stomatitis virus is enhanced by expression of the fusion cytosine deaminase/uracil phosphoribosyltransferase suicide gene. Cancer Res. 63:8366–8376. 2003.
15	Sun X, Xing L, Deng X, et al: Hypoxia targeted bifunctional suicide gene expression enhances radiotherapy in vitro and in vivo. Radiother Oncol. 105:57–63. 2012. View Article : Google Scholar : PubMed/NCBI
16	Xing L, Sun X, Deng X, et al: A triple suicide gene strategy that improves therapeutic effects and incorporates multimodality molecular imaging for monitoring gene functions. Cancer Gene Therapy. 20:358–365. 2013. View Article : Google Scholar : PubMed/NCBI
17	He F, Deng X, Wen B, et al: Noninvasive molecular imaging of hypoxia in human xenografts: comparing hypoxia-induced gene expression with endogenous and exogenous hypoxia markers. Cancer Res. 68:8597–8606. 2008. View Article : Google Scholar : PubMed/NCBI
18	Binley K, Askham Z, Martin L, et al: Hypoxia-mediated tumour targeting. Gene Ther. 10:540–549. 2003. View Article : Google Scholar : PubMed/NCBI
19	Liu J, Harada H, Ogura M, Shibata T and Hiraoka M: Adenovirus-mediated hypoxia-targeting cytosine deaminase gene therapy enhances radiotherapy in tumour xenografts. Br J Cancer. 96:1871–1878. 2007. View Article : Google Scholar : PubMed/NCBI
20	Lee CH, Wu CL and Shiau AL: Hypoxia-induced cytosine deaminase gene expression for cancer therapy. Hum Gene Ther. 18:27–38. 2007. View Article : Google Scholar : PubMed/NCBI
21	Wang D, Ruan H, Hu L, et al: Development of a hypoxia-inducible cytosine deaminase expression vector for gene-directed prodrug cancer therapy. Cancer Gene Ther. 12:276–283. 2005. View Article : Google Scholar : PubMed/NCBI
22	Chen JK, Hu LJ, Wang D, Lamborn KR and Deen DF: Cytosine deaminase/5-fluorocytosine exposure induces bystander and radiosensitization effects in hypoxic glioblastoma cells in vitro. Int J Radiat Oncol Biol Phys. 67:1538–1547. 2007. View Article : Google Scholar : PubMed/NCBI
23	Freytag SO, Kim JH, Brown SL, Barton K, Lu M and Chung M: Gene therapy strategies to improve the effectiveness of cancer radiotherapy. Expert Opin Biol Ther. 4:1757–1770. 2004. View Article : Google Scholar : PubMed/NCBI
24	Hwang HS, Davis TW, Houghton JA and Kinsella TJ: Radiosensitivity of thymidylate synthase-deficient human tumor cells is affected by progression through the G1 restriction point into S-phase: implications for fluoropyrimidine radiosensitization. Cancer Res. 60:92–100. 2000.
25	McMasters RA, Saylors RL, Jones KE, Hendrix ME, Moyer MP and Drake RR: Lack of bystander killing in herpes simplex virus thymidine kinase-transduced colon cell lines due to deficient connexin43 gap junction formation. Hum Gen Ther. 9:2253–2261. 1998. View Article : Google Scholar
26	Shirakawa T, Gardner TA, Ko SC, et al: Cytotoxicity of adenoviral-mediated cytosine deaminase plus 5-fluorocytosine gene therapy is superior to thymidine kinase plus acyclovir in a human renal cell carcinoma model. J Urol. 162:949–954. 1999. View Article : Google Scholar : PubMed/NCBI
27	Khatri A, Zhang B, Doherty E, et al: Combination of cytosine deaminase with uracil phosphoribosyl transferase leads to local and distant bystander effects against RM1 prostate cancer in mice. J Gene Med. 8:1086–1096. 2006. View Article : Google Scholar : PubMed/NCBI