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Cited by 16Virus-based gene therapy researchCancer, Hypoxia, and MetabolismAngiogenesis and VEGF in Cancer

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Stable knockdown of CREB, HIF-1 and HIF-2 by replication-competent retroviruses abrogates the responses to hypoxia in hepatocellular carcinoma

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Shneor, D, Folberg, R, Pe'er, J, Honigman, A, Frenkel, S·2017·Cancer Gene Therapy
DOI 10.1038/cgt.2016.68PMC PMC5339434
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Abstract

The fast proliferation of tumor cells develops faster than the vasculature, resulting, in most malignant tumors, in generation of hypoxic regions. Hypoxia renders solid tumors resistant to radiation and chemotherapeutics while providing opportunities for tumor-selective therapies targeting tumor hypoxia. Here we exploit two properties of tumors: propagation of tumor cells and ongoing generation of hypoxic regions to construct a system that preferentially leads to the death of tumor cells and thus hinders tumor growth. We constructed murine leukemia virus replication-competent (RCR) viruses that infect only propagating cells. These viruses express small hairpin RNAs (shRNAs) targeting cyclic AMP-response-element binding protein (CREB), hypoxia-inducible factors 1 (HIF)-1 or HIF-2 individually or all three together (X3). These viruses efficiently infected in vitro human hepatocellular carcinoma (HepG2 and FLC4) cells and established persistence of the virus and knocked down the expression of the regulators of the hypoxia-responding genes. Knockdown of either HIF-1 or CREB or both in hypoxia reduced the expression of hypoxia-response elements- and CRE-mediated gene expression, diminished cell proliferation and increased caspase-3 activity. We did not detect any significant effect of the efficiently knocked down HIF-2 on any of the functions tested in vitro. Moreover, severe combined immunodeficiency mice implanted subcutaneously with HepG2 stably infected with recombinant RCRs showed reduction of tumor growth and vascular endothelial growth factor expression, and no hypoxia-guided neovascularization. Combined treatment (RCRs+doxorubicin) improved efficacy in the context of in vitro hypoxia and in vivo (with either vACE-CREB or vACE-X3). This synergistic effect may lead to an improved efficacy and safety profile of the treatment that may result in fewer side effects.

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Figure 1 A schematic presentation of the various replication-competent viruses (RCRs). (a) A proviral construct of pACE-GFP. (b–f) The IRES-GFP sequences were replaced with an H1 promoter driving the transcription of the various small hairpin RNAs (shRNA) targeting cyclic AMP-response-element binding protein (CREB) (b), hypoxia-inducible factor (HIF)-1 (c), HIF-2 (d), all three genes (e) and a non-target sequence (f). The DNA sequence of the various shRNAs is marked.
Figure 1 A schematic presentation of the various replication-competent viruses (RCRs). (a) A proviral construct of pACE-GFP. (b–f) The IRES-GFP sequences were replaced with an H1 promoter driving the transcription of the various small hairpin RNAs (shRNA) targeting cyclic AMP-response-element binding protein (CREB) (b), hypoxia-inducible factor (HIF)-1 (c), HIF-2 (d), all three genes (e) and a non-target sequence (f). The DNA sequence of the various shRNAs is marked.CC BY-NC-ND · PubMed Central
Figure 2 Quantification of the efficiency of the knockdown in infected cell lines and the effect on the expression of regulated genes. Cells, HepG2 and FLC4 were infected with each of the recombinant replication-competent retroviruses (RCRs) as specified in the figure and cultured in hypoxic conditions for 24 h. Purified mRNA was quantified by real-time quantitative PCR. Levels of messenger RNA (mRNA) were normalized to mRNAs in cells infected with vACE-NT (set as 100%) (a–c). Proteins were quantified following western blot analysis (g), and band intensities were compared with those of α-tubulin and to band intensity of cells infected with vACE-NT (set as 100%) (d–f). Levels of all measured mRNAs (a–c) and proteins (d–f) were statistically significantly lower when compared with vACE-NT-infected cells in both cell lines (P<0.05). HepG2 cells infected with the various RCRs were transiently transfected with either pCRE-LUC or pHRE-LUC. Luciferase activity was determined after 48 h of exposure to hypoxia relative to cells infected with vACE-NT (presented as 100%) for each transfection. CRE (dark columns) and hypoxia-response elements (HRE) (light columns) mediated LUC activity are presented in relative light units (P<0.001) (h). The levels of vascular endothelial growth factor (VEGF) secreted by HepG2 cells were determined by enzyme-linked immunosorbent assays as described in the Materials and methods section. The VEGF secretion levels in hypoxia relative to normoxia were normalized to the ratio in vACE-NT-infected cells (P<0.01) (i). α-Tub=α-tubulin. Bars represent mean+s.d. for each measurement.
Figure 2 Quantification of the efficiency of the knockdown in infected cell lines and the effect on the expression of regulated genes. Cells, HepG2 and FLC4 were infected with each of the recombinant replication-competent retroviruses (RCRs) as specified in the figure and cultured in hypoxic conditions for 24 h. Purified mRNA was quantified by real-time quantitative PCR. Levels of messenger RNA (mRNA) were normalized to mRNAs in cells infected with vACE-NT (set as 100%) (a–c). Proteins were quantified following western blot analysis (g), and band intensities were compared with those of α-tubulin and to band intensity of cells infected with vACE-NT (set as 100%) (d–f). Levels of all measured mRNAs (a–c) and proteins (d–f) were statistically significantly lower when compared with vACE-NT-infected cells in both cell lines (P<0.05). HepG2 cells infected with the various RCRs were transiently transfected with either pCRE-LUC or pHRE-LUC. Luciferase activity was determined after 48 h of exposure to hypoxia relative to cells infected with vACE-NT (presented as 100%) for each transfection. CRE (dark columns) and hypoxia-response elements (HRE) (light columns) mediated LUC activity are presented in relative light units (P<0.001) (h). The levels of vascular endothelial growth factor (VEGF) secreted by HepG2 cells were determined by enzyme-linked immunosorbent assays as described in the Materials and methods section. The VEGF secretion levels in hypoxia relative to normoxia were normalized to the ratio in vACE-NT-infected cells (P<0.01) (i). α-Tub=α-tubulin. Bars represent mean+s.d. for each measurement.CC BY-NC-ND · PubMed Central
Figure 3 The role of cyclic AMP-response-element binding protein (CREB), hypoxia-inducible factor (HIF)-1 and HIF-2 in protecting HepG2 and FLC4 cells from hypoxia-induced apoptosis and on the response to treatment with doxorubicin (DOX). HepG2 (a, c) and FLC4 (b, d) infected with each of the viruses specified in the figure were cultured for 72 h. Viability (a, b) and activation of caspase-3 (Cas3) (c, d) were determined every 24 h. The relative viability and activation of caspase-3 after 72 h of hypoxia (light gray columns), and those after treatment with 1 mm of DOX in normoxia (black columns) and hypoxia (dark gray columns) were normalized to cells infected with each respective replication competent retroviruses in normoxia (P<0.01 for all comparisons). Bars represent mean+s.d. for each measurement.
Figure 3 The role of cyclic AMP-response-element binding protein (CREB), hypoxia-inducible factor (HIF)-1 and HIF-2 in protecting HepG2 and FLC4 cells from hypoxia-induced apoptosis and on the response to treatment with doxorubicin (DOX). HepG2 (a, c) and FLC4 (b, d) infected with each of the viruses specified in the figure were cultured for 72 h. Viability (a, b) and activation of caspase-3 (Cas3) (c, d) were determined every 24 h. The relative viability and activation of caspase-3 after 72 h of hypoxia (light gray columns), and those after treatment with 1 mm of DOX in normoxia (black columns) and hypoxia (dark gray columns) were normalized to cells infected with each respective replication competent retroviruses in normoxia (P<0.01 for all comparisons). Bars represent mean+s.d. for each measurement.CC BY-NC-ND · PubMed Central
Figure 4 Testing the effect of cyclic AMP-response-element binding protein (CREB), HIF-1 and 2 on tumor growth in vivo in a mouse xenograft model. Severe combined immunodeficiency mice were implanted with 4 × 106 HepG2 cells as described in Materials and methods section. Tumor growth was monitored every 7 days with the IVIS InVivo System. Images of light emission from representative mice taken on days 7 and 35 are presented. The same color scale was used for all mice at both time points (a). Relative light units (RLU) measurements were normalized to the readings on day 7 post injection and plotted along the extent of the experiment. Each point represents the mean+s.e. (b). At the end of the experiment, tumors were harvested for histopathological analysis. Hematoxylin–eosin staining and the corresponding vascular endothelial growth factor immunohistochemistry of tumors are presented in (c). ( × 4 magnification, scale bar 100 μm).
Figure 4 Testing the effect of cyclic AMP-response-element binding protein (CREB), HIF-1 and 2 on tumor growth in vivo in a mouse xenograft model. Severe combined immunodeficiency mice were implanted with 4 × 106 HepG2 cells as described in Materials and methods section. Tumor growth was monitored every 7 days with the IVIS InVivo System. Images of light emission from representative mice taken on days 7 and 35 are presented. The same color scale was used for all mice at both time points (a). Relative light units (RLU) measurements were normalized to the readings on day 7 post injection and plotted along the extent of the experiment. Each point represents the mean+s.e. (b). At the end of the experiment, tumors were harvested for histopathological analysis. Hematoxylin–eosin staining and the corresponding vascular endothelial growth factor immunohistochemistry of tumors are presented in (c). ( × 4 magnification, scale bar 100 μm).CC BY-NC-ND · PubMed Central
Figure 5 Immunohistochemical analysis of hypoxia and blood vessels (CD34) of the tumors. Tumors infected with replication-competent retroviruses expressing small hairpin RNA (shRNA)-targeting genes specified on the left column were stained for hypoxia (left column) and for blood vessels (right column) as described in the Materials and methods section. It is clear that knockdown of CREB, HIF-1 and all three genes together ( × 3) abolished neovascularization within the tumors. In tumors infected with the non-target shRNA or with shRNA targeting HIF-2 blood vessel growth toward the hypoxic regions is noticed. Correlation between the amount of CD34 and hypoxia staining is presented in the top right column for each virus. ( × 10 magnification, scale bar 500 μm).
Figure 5 Immunohistochemical analysis of hypoxia and blood vessels (CD34) of the tumors. Tumors infected with replication-competent retroviruses expressing small hairpin RNA (shRNA)-targeting genes specified on the left column were stained for hypoxia (left column) and for blood vessels (right column) as described in the Materials and methods section. It is clear that knockdown of CREB, HIF-1 and all three genes together ( × 3) abolished neovascularization within the tumors. In tumors infected with the non-target shRNA or with shRNA targeting HIF-2 blood vessel growth toward the hypoxic regions is noticed. Correlation between the amount of CD34 and hypoxia staining is presented in the top right column for each virus. ( × 10 magnification, scale bar 500 μm).CC BY-NC-ND · PubMed Central
Figure 6 Effect of combined treatment on tumor growth monitored by light emission. HepG2 cells stably expressing the luc gene and infected with the viruses specified above were implanted subcutaneous in severe combined immunodeficiency (SCID) mice. After 3 weeks, the mice were injected twice a week with doxorubicin (DOX) in the concentrations specified in the picture. Tumor growth was monitored by light emission as described in Figure 4. (a) One mouse from each group at day 42 is depicted. A scale bar of light emission is localized on the right side of the picture. (b) The average (+s.d.) relative tumor growth rate between days 14 and 42 is presented on each bar.
Figure 6 Effect of combined treatment on tumor growth monitored by light emission. HepG2 cells stably expressing the luc gene and infected with the viruses specified above were implanted subcutaneous in severe combined immunodeficiency (SCID) mice. After 3 weeks, the mice were injected twice a week with doxorubicin (DOX) in the concentrations specified in the picture. Tumor growth was monitored by light emission as described in Figure 4. (a) One mouse from each group at day 42 is depicted. A scale bar of light emission is localized on the right side of the picture. (b) The average (+s.d.) relative tumor growth rate between days 14 and 42 is presented on each bar.CC BY-NC-ND · PubMed Central

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Citations

Cites

Research works printed but not linked: 49

  1. Dhanasekaran R, Limaye A, Cabrera R . Hepatocellular carcinoma: current trends in worldwide epidemiology, risk factors, diagnosis, and therapeutics. Hepat Med 2012; 4: 19–37.
  2. Abramovitch R, Tavor E, Jacob-Hirsch J, Zeira E, Amariglio N, Pappo O et al. A pivotal role of cyclic AMP-responsive element binding protein in tumor progression. Cancer Res 2004; 64: 1338–1346. · DOI
  3. Lu X, Kang Y . Hypoxia and hypoxia-inducible factors: master regulators of metastasis. Clin Cancer Res 2010; 16: 5928–5935. · DOI · Paper page
  4. Rhim T, Lee DY, Lee M . Hypoxia as a target for tissue specific gene therapy. J Control Release 2013; 172: 484–494. · DOI
  5. Durand RE, Sham E . The lifetime of hypoxic human tumor cells. Int J Radiat Oncol Biol Phys 1998; 42: 711–715. · DOI
  6. Wu XZ, Xie GR, Chen D . Hypoxia and hepatocellular carcinoma: the therapeutic target for hepatocellular carcinoma. J Gastroenterol Hepatol 2007; 22: 1178–1182. · DOI
  7. Harris AL . Hypoxia—a key regulatory factor in tumour growth. Nat Rev Cancer 2002; 2: 38–47. · DOI · Paper page
  8. Le QT, Denko NC, Giaccia AJ . Hypoxic gene expression and metastasis. Cancer Metastasis Rev 2004; 23: 293–310. · DOI
  9. Majmundar AJ, Wong WJ, Simon MC . Hypoxia-inducible factors and the response to hypoxic stress. Mol Cell 2010; 40: 294–309. · DOI · Paper page
  10. Choi SH, Shin HW, Park JY, Yoo JY, Kim do Y, Ro WS et al. Effects of the knockdown of hypoxia inducible factor-1 alpha expression by adenovirus-mediated shRNA on angiogenesis and tumor growth in hepatocellular carcinoma cell lines. Korean J Hepatol 2010; 16: 280–287. · DOI
  11. Holmquist-Mengelbier L, Fredlund E, Lofstedt T, Noguera R, Navarro S, Nilsson H et al. Recruitment of HIF-1alpha and HIF-2alpha to common target genes is differentially regulated in neuroblastoma: HIF-2alpha promotes an aggressive phenotype. Cancer Cell 2006; 10: 413–423. · DOI · Paper page
  12. White PC, Shore AM, Clement M, McLaren J, Soeiro I, Lam EW et al. Regulation of cyclin D2 and the cyclin D2 promoter by protein kinase A and CREB in lymphocytes. Oncogene 2006; 25: 2170–2180. · DOI
  13. Mayr B, Montminy M . Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol 2001; 2: 599–609. · DOI · Paper page
  14. Aggarwal S, Kim SW, Ryu SH, Chung WC, Koo JS . Growth suppression of lung cancer cells by targeting cyclic AMP response element-binding protein. Cancer Res 2008; 68: 981–988. · DOI
  15. Belkhiri A, Dar AA, Zaika A, Kelley M, El-Rifai W . t-Darpp promotes cancer cell survival by up-regulation of Bcl2 through Akt-dependent mechanism. Cancer Res 2008; 68: 395–403. · DOI
  16. Signorelli S, Jennings P, Leonard MO, Pfaller W . Differential effects of hypoxic stress in alveolar epithelial cells and microvascular endothelial cells. Cell Physiol Biochem 2010; 25: 135–144. · DOI
  17. Park SI, Park SJ, Lee J, Kim HE, Sohn JW, Park YG . Inhibition of cyclic AMP response element-directed transcription by decoy oligonucleotides enhances tumor-specific radiosensitivity. Biochem Biophys Res Commun 2016; 469: 363–369. · DOI
  18. Kovach SJ, Price JA, Shaw CM, Theodorakis NG, McKillop IH . Role of cyclic-AMP responsive element binding (CREB) proteins in cell proliferation in a rat model of hepatocellular carcinoma. J Cell Physiol 2006; 206: 411–419. · DOI
  19. Yu L, Guo X, Zhang P, Qi R, Li Z, Zhang S . Cyclic adenosine monophosphate-responsive element-binding protein activation predicts an unfavorable prognosis in patients with hepatocellular carcinoma. OncoTargets Ther 2014; 7: 873–879.
  20. Block A, Freund CT, Chen SH, Nguyen KP, Finegold M, Windler E et al. Gene therapy of metastatic colon carcinoma: regression of multiple hepatic metastases by adenoviral expression of bacterial cytosine deaminase. Cancer Gene Ther 2000; 7: 438–445. · DOI
  21. Chen SH, Chen XH, Wang Y, Kosai K, Finegold MJ, Rich SS et al. Combination gene therapy for liver metastasis of colon carcinoma in vivo. Proc Natl Acad Sci USA 1995; 92: 2577–2581. · DOI
  22. Nyati MK, Symon Z, Kievit E, Dornfeld KJ, Rynkiewicz SD, Ross BD et al. The potential of 5-fluorocytosine/cytosine deaminase enzyme prodrug gene therapy in an intrahepatic colon cancer model. Gene Ther 2002; 9: 844–849. · DOI
  23. Zhang M, Li S, Nyati MK, DeRemer S, Parsels J, Rehemtulla A et al. Regional delivery and selective expression of a high-activity yeast cytosine deaminase in an intrahepatic colon cancer model. Cancer Res 2003; 63: 658–663.
  24. Sung MW, Yeh HC, Thung SN, Schwartz ME, Mandeli JP, Chen SH et al. Intratumoral adenovirus-mediated suicide gene transfer for hepatic metastases from colorectal adenocarcinoma: results of a phase I clinical trial. Mol Ther 2001; 4: 182–191. · DOI
  25. Logg CR, Robbins JM, Jolly DJ, Gruber HE, Kasahara N . Retroviral replicating vectors in cancer. Methods Enzymol 2012; 507: 199–228. · DOI
  26. Roe T, Reynolds TC, Yu G, Brown PO . Integration of murine leukemia virus DNA depends on mitosis. EMBO J 1993; 12: 2099–2108. · DOI · Paper page
  27. Miller DG, Adam MA, Miller AD . Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol 1990; 10: 4239–4242. · DOI
  28. Hiraoka K, Kimura T, Logg CR, Tai CK, Haga K, Lawson GW et al. Therapeutic efficacy of replication-competent retrovirus vector-mediated suicide gene therapy in a multifocal colorectal cancer metastasis model. Cancer Res 2007; 67: 5345–5353. · DOI
  29. Tai CK, Kasahara N . Replication-competent retrovirus vectors for cancer gene therapy. Front Biosci 2008; 13: 3083–3095. · DOI
  30. Logg CR, Logg A, Matusik RJ, Bochner BH, Kasahara N . Tissue-specific transcriptional targeting of a replication-competent retroviral vector. J Virol 2002; 76: 12783–12791. · DOI
  31. Logg CR, Tai CK, Logg A, Anderson WF, Kasahara N . A uniquely stable replication-competent retrovirus vector achieves efficient gene delivery in vitro and in solid tumors. Hum Gene Ther 2001; 12: 921–932. · DOI
  32. Sliva K, Schnierle BS . Selective gene silencing by viral delivery of short hairpin RNA. Virol J 2010; 7: 248. · DOI
  33. McIntyre GJ, Arndt AJ, Gillespie KM, Mak WM, Fanning GC . A comparison of multiple shRNA expression methods for combinatorial RNAi. Genet Vaccines Ther 2011; 9: 9. · DOI
  34. Sliva K, Schnierle BS . Stable integration of a functional shRNA expression cassette into the murine leukemia virus genome. Virology 2006; 351: 218–225. · DOI
  35. Li T, Liu Z, Jiang K, Ruan Q . Angiopoietin2 enhances doxorubicin resistance in HepG2 cells by upregulating survivin and Ref-1 via MSK1 activation. Cancer Lett 2013; 337: 276–284. · DOI
  36. Sayan M, Shukla A, MacPherson MB, Macura SL, Hillegass JM, Perkins TN et al. Extracellular signal-regulated kinase 5 and cyclic AMP response element binding protein are novel pathways inhibited by vandetanib (ZD6474) and doxorubicin in mesotheliomas. Am J Respir Cell Mol Biol 2014; 51: 595–603. · DOI
  37. Gilliam LA, Moylan JS, Ann Callahan L, Sumandea MP, Reid MB . Doxorubicin causes diaphragm weakness in murine models of cancer chemotherapy. Muscle Nerve 2011; 43: 94–102. · DOI
  38. Yang ZF, Poon RT . Vascular changes in hepatocellular carcinoma. Anat Rec 2008; 291: 721–734. · DOI
  39. Ryan HE, Lo J, Johnson RS . HIF-1 alpha is required for solid tumor formation and embryonic vascularization. EMBO J 1998; 17: 3005–3015. · DOI · Paper page
  40. Jeon SH, Chae BC, Kim HA, Seo GY, Seo DW, Chun GT et al. The PKA/CREB pathway is closely involved in VEGF expression in mouse macrophages. Mol Cells 2007; 23: 23–29. · DOI
  41. Zhu AX, Duda DG, Sahani DV, Jain RK . HCC and angiogenesis: possible targets and future directions. Nat Rev Clin Oncol 2011; 8: 292–301. · DOI
  42. Kizaka-Kondoh S, Inoue M, Harada H, Hiraoka M . Tumor hypoxia: a target for selective cancer therapy. Cancer Sci 2003; 94: 1021–1028. · DOI
  43. Tai CK, Wang W, Lai YH, Logg CR, Parker WB, Li YF et al. Enhanced efficiency of prodrug activation therapy by tumor-selective replicating retrovirus vectors armed with the Escherichia coli purine nucleoside phosphorylase gene. Cancer Gene Ther 2010; 17: 614–623. · DOI
  44. Tai CK, Wang WJ, Chen TC, Kasahara N . Single-shot, multicycle suicide gene therapy by replication-competent retrovirus vectors achieves long-term survival benefit in experimental glioma. Mol Ther 2005; 12: 842–851. · DOI
  45. Wang WJ, Tai CK, Kasahara N, Chen TC . Highly efficient and tumor-restricted gene transfer to malignant gliomas by replication-competent retroviral vectors. Hum Gene Ther 2003; 14: 117–127. · DOI
  46. Takahashi M, Valdes G, Hiraoka K, Inagaki A, Kamijima S, Micewicz E et al. Radiosensitization of gliomas by intracellular generation of 5-fluorouracil potentiates prodrug activator gene therapy with a retroviral replicating vector. Cancer Gene Ther 2014; 21: 405–410. · DOI
  47. Huang TT, Hlavaty J, Ostertag D, Espinoza FL, Martin B, Petznek H et al. Toca 511 gene transfer and 5-fluorocytosine in combination with temozolomide demonstrates synergistic therapeutic efficacy in a temozolomide-sensitive glioblastoma model. Cancer Gene Ther 2013; 20: 544–551. · DOI
  48. He C, Sun XP, Qiao H, Jiang X, Wang D, Jin X et al. Downregulating hypoxia-inducible factor-2alpha improves the efficacy of doxorubicin in the treatment of hepatocellular carcinoma. Cancer Sci 2012; 103: 528–534. · DOI
  49. Ye CG, Yeung JH, Huang GL, Cui P, Wang J, Zou Y et al. Increased glutathione and mitogen-activated protein kinase phosphorylation are involved in the induction of doxorubicin resistance in hepatocellular carcinoma cells. Hepatol Res 2013; 43: 289–299. · DOI

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Metadata

Authors
Shneor, D., Folberg, R., Pe'er, J., Honigman, A., & Frenkel, S.
Affiliations
  • Hebrew University of Jerusalem (IL)
  • Oakland University (US)
  • Hadassah Medical Center (IL)
Corresponding author
Frenkel, S
ORCID
Venue
Cancer Gene Therapy
Publisher
Springer Science and Business Media LLC
Published
2016-12-09
Volume
24
Issue
2
Pages
64-74
ISSN
  • 0929-1903 (ISSN-L)
  • 1476-5500
DOI
10.1038/cgt.2016.68
PMC
PMC5339434
OpenAlex
W2560657110
Cited by
16
References
4
Citations by year
2026: 1 · 2024: 1 · 2023: 3 · 2021: 2 · 2020: 2 · 2019: 5 · 2018: 1 · 2017: 1
FWCI
1.89
Type
journal article
Language
en
Open access
Licence
Locations
  • Cancer Gene Therapy (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Hebrew University of Jerusalem
  • Israel Cancer Association
  • Israel Science Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
Keywords
hepatocellular carcinoma, tumor hypoxia, CREB, HIF-1, HIF-2, murine leukemia virus, shRNA, gene knockdown, vascular endothelial growth factor, tumor angiogenesis, doxorubicin combination therapy, caspase-3, tumor growth inhibition
MeSH terms
Endothelial PAS Domain-Containing Protein 1, Animals, Hypoxia, Disease Models, Animal, Genetic Vectors, Carcinoma, Hepatocellular, Humans, Liver Neoplasms, Neovascularization, Pathologic, Retroviridae, Biomarkers, Gene Expression Regulation, Neoplastic, Cyclic AMP Response Element-Binding Protein, Xenograft Model Antitumor Assays, Gene Order, RNA, Small Interfering, Cell Line, Tumor, Tumor Burden, Cell Proliferation, Mice, Basic Helix-Loop-Helix Transcription Factors, Basic Helix-Loop-Helix Proteins, Hypoxia-Inducible Factor 1, Gene Knockdown Techniques

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