Hyperoxia retards growth and induces apoptosis, changes in vascular density and gene expression in transplanted gliomas in nude rats | springermedicine.com

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Journal of Neuro-Oncology

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01-11-2007 | lab Invastigation-human/animal tissue

Hyperoxia retards growth and induces apoptosis, changes in vascular density and gene expression in transplanted gliomas in nude rats

Authors: Linda Elin Birkhaug Stuhr, A. Raa, A. M. Øyan, K. H. Kalland, P. O. Sakariassen, K. Petersen, R. Bjerkvig, R. K. Reed

Published in: Journal of Neuro-Oncology | Issue 2/2007

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Abstract

This study describes the biological effects of hyperoxic treatment on BT4C rat glioma xenografts in vivo with special reference to tumor growth, angiogenesis, apoptosis, general morphology and gene expression parameters.

One group of tumor bearing animals was exposed to normobaric hyperoxia (1 bar, pO₂ = 1.0) and another group was exposed to hyperbaric hyperoxia (2 bar, pO₂ = 2.0), whereas animals housed under normal atmosphere (1 bar, pO₂ = 0.2) served as controls. All treatments were performed at day 1, 4 and 7 for 90 min. Treatment effects were determined by assessment of tumor growth, vascular morphology (immunostaining for von Willebrand factor), apoptosis by TUNEL staining and cell proliferation by Ki67 staining. Moreover, gene expression profiles were obtained and verified by real time quantitative PCR.

Hyperoxic treatment caused a ∼60% reduction in tumor growth compared to the control group after 9 days (p < 0.01). Light microscopy showed that the tumors exposed to hyperoxia contained large “empty spaces” within the tumor mass. Moreover, hyperoxia induced a significant increase in the fraction of apoptotic cells (∼21%), with no significant change in cell proliferation. After 2 bar treatment, the mean vascular density was reduced in the central parts of the tumors compared to the control and 1 bar group. The vessel diameters were significantly reduced (11–24%) in both parts of the tumor tissue. Evidence of induced cell death and reduced angiogenesis was reflected by gene expression analyses.

Increased pO₂−levels in experimental gliomas, using normobaric and moderate hyperbaric oxygen therapy, caused a significant reduction in tumor growth. This process is characterized by enhanced cell death, reduced vascular density and changes in gene expression corresponding to these effects.

1.

Vaupel P, Kallinowski F, Okunieff P (1989) Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res 1:6449–6465 (Review)

2.

Höckel M, Vaupel P (2001) Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. J Natl Cancer Inst 21:266–276 (Review)CrossRef

3.

Brizel DM, Lin S, Johnson JL, Brooks J, Dewhirst MW, Piantadosi CA (1995) The mechanisms by which hyperbaric oxygen and carbogen improve tumor oxygenation. Br J Cancer 72:1120–1124PubMed

4.

Semenza GL (2000) HIF-1 and human disease: one highly involved factor. Genes Dev 15:1983–1991 (Review)

5.

Helczynska K, Kronblad A, Jogi A, Nilsson E, Beckman S, Landberg G, Pahlman S (2003) Hypoxia promotes a dedifferentiated phenotype in ductal breast carcinoma in situ. Cancer Res 1:1441–1444

6.

Beppu T, Kamada K, Yoshida Y, Arai H, Ogasawara K, Ogawa A (2002) Change of oxygen pressure in glioblastoma tissue under various conditions. J Neurooncol 58:47–52PubMedCrossRef

7.

Becker A, Kuhnt T, Liedtke H, Krivokuca A, Bloching M, Dunst J (2002) Oxygenation measurements in head and neck cancers during hyperbaric oxygenation. Strahlenther Onkol 178:105–108PubMedCrossRef

8.

Kinoshita Y, Kohshi K, Kunugita N, Tosakki T, Yokota A (2000) Preservation of tumor oxygen after hyperbaric oxygenation monitored by magnetic resonance imagin. Br J Cancer 82:88–92PubMedCrossRef

9.

Al-Waili NS, Butler GJ, Beale J, Hamilton RW, Lee BY, Lucas P (2005) Hyperbaric oxygen and malignancies: a potential role in radiotherapy, chemotherapy, tumor surgery and phototherapy. Med Sci Monit 11:279–289

10.

Kohshi K, Kinoshita Y, Imada H, Kunuqita N, Abe H, Terashima H, Tokui N, Vemura S (1999) Effects of radiotherapy after hyperbaric oxygenation on malignant gliomas. Br J Cancer 80:236–241PubMedCrossRef

11.

Ogawa K, Yoshii Y, Inoue O, Toita T, Saito A, Kakinohana Y, Adachi G, Ishikawa Y, Kin S, Murayama S (2003) Prospective trial of radiotherapy after hyperbaric oxygenation with chemotherapy for high-grade gliomas. Radiother Oncol 67:63–67PubMedCrossRef

12.

Ogawa K, Yoshii Y, Inoue O, Toita T, Saito A, Kakinohana Y, Adachi G, Iraha S, Tamaki W, Sugimoto K, Hyodo A, Murayama S (2006) Phase II trial of radiotherapy after hyperbaric oxygenation with chemotherapy for high-grade gliomas. Br J Cancer 9:862–868CrossRef

13.

Feldmeier JJ, Hampson NB (2002) A systemic review of the literature reporting the application of hyperbaric oxygen prevention and treatment of delayed radiation injury:an evidence-based approach. Undersa Hyperbar Med 29:4–30

14.

Gill AL, Bell CAN (2004) Hyperbaric oxygen: its uses, mechanisms of action and outcomes. Q J Med 97:385–395

15.

Siemann DW, Warrington KH, Horsman MR (2000) Targeting tumor blood vessels: an adjuvant strategy for radiation therapy. Radiother Oncol 57:5–12 (Review)PubMedCrossRef

16.

Jamieson D, van den Bronk HAS (1963) Measurements of oxygen tension in cerebral tissues of rats exposed to high pressure of oxygen. J Appl Phsyiol 18:869–876

17.

Stuhr LE, Iversen VV, Straume O, Maehle BO, Reed RK (2004) Hyperbaric oxygen alone or combined with 5-FU attenuates growth of DMBA-induced rat mammary tumors Cancer Lett 210:35–40PubMedCrossRef

18.

Raa A, Stansberg C, Steen VM, Bjerkvig R, Reed RK, Stuhr LEB (2007) Hyperoxia retards growth and induces apoptosis and loss of glands and blood vessels in DMBA-induced rat mammary tumors. BMC Cancer 7:23 (Epub ahead of print)PubMedCrossRef

19.

McDonald KR (1996) Effect of hyperbaric oxygenation on existing oral mucosal carcinoma. Laryngoscope 106:957–959PubMedCrossRef

20.

Lian QL (1995) Effects of hyperbaric oxygen on S-180 sarcoma in mice. Undersea Hyperbar med 22:153–160

21.

Laerum OD, Rajewsky MF, Schachner M, Stavrou D, Haglid KH, Haugen Å (1977) Phenotypic properties of neoplastic cell lines developed from fetal rat brain cells in culture after exposure to ethylnitrosourea in vivo. Z. Krebsforsch 89:273–295CrossRef

22.

Sandstrom M, Johansson M, Andersson U, Bergh A, Bergenheim AT, Henriksson R (2004) Related Articles, Links The tyrosine kinase inhibitor ZD6474 inhibits tumor growth in an intracerebral rat glioma model. Br J Cancer 91:1174–1180PubMed

23.

Thorsen F, Ersland L, Nordli H, Enger PO, Huszthy PC, Lundervold A, Standnes T, Bjerkvig R, Lund-Johansen M (2003) Imaging of experimental rat gliomas using a clinical MR scanner. J Neurooncol 63:225–231PubMedCrossRef

24.

Vallbo C, Bergenheim T, Hedman H, Henriksson R (2002) The antimicrotubule drug estramustine but not irradiation induces apoptosis in malignant glioma involving AKT and caspase pathways. J Neurooncol 56:143–148PubMedCrossRef

25.

Rostad K, Mannelqvist M, Halvorsen OJ, Oyan AM, Bo TH, Stordrange L, Olsen S, Haukaas SA, Lin B, Hood L, Jonassen I, Akslen LA, Kalland KH (2007) ERG upregulation and related ETS transcription factors in prostate cancer. Int J Oncol 30:19–32PubMed

26.

Anensen N, Oyan AM, Bourdon JC, Kalland KH, Bruserud O, Gjertsen BT (2006) A distinct p53 protein isoform signature reflects the onset of induction chemotherapy for acute myeloid leukemia. Clin Cancer Res 12:3985–3992PubMedCrossRef

27.

Dysvik B, Jonassen I (2001) J-Express: exploring gene expression data wing Java. Bioinfomatics 17:369–370CrossRef

28.

Yang YH, Buckley MJ, Speed TP (2001) Analysis of cDNA microarray images. Briefings in Bioinformatics 2:341–349PubMedCrossRef

29.

Bolstad BM, Irizarry RA, Astrand M, Speed TP (2003) A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinfomatics 19:185–193CrossRef

30.

Margaretten NC, Witschi H (1988) Effects of hyperoxia on growth characteristics of metastatic murine tumors in the lung. Cancer Res 15:2779–2783

31.

Lindenschmidt RC, Margaretten N, Griesemer RA, Witschi HP (1986) Modification of lung tumor growth by hyperoxia. Carcinogenesis 7:1581–1586PubMedCrossRef

32.

Clark JM (1993) Oxygen toxicity. In: Bennet P, Elliot D (eds) The physiology and medicine of diving, 4th edn. WB Saunders Company Ltd, 121–169

33.

McMillan T, Calhoun KH, Mader JT, Stiernberg CM, Rajaraman S (1989) The effect of hyperbaric oxygen on oral mucosal carcinoma. Laryngoscop 99:241–244

34.

Daruwalla J, Christophi C (2006) Hyperbaric oxygen therapy for malignancy: a review. World J Surg, e-publish ahead of print nov 7

35.

Folkman J (1995) Clinical application of research on angiogenesis. N Engl J Med 333:1757–1763PubMedCrossRef

36.

Hanahan D, Folkman J (1996) Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 863:353–364CrossRef

37.

Shih AH, Holland EC (2006) Platelet-derived growth factor (PDGF) and glial tumorigenesis. Cancer Lett 232:139–147PubMedCrossRef

38.

Ko YJ, Small EJ, Kabbinavar F, Chachoua A, Taneja S, Reese D, DePaoli A, Hannah A, Balk SP, Bubley GJ (2001) A multi-institutional phase ii study of SU101, a platelet-derived growth factor receptor inhibitor, for patients with hormone-refractory prostate cancer. Clin Cancer Res 7:800–805PubMed

39.

Eckhardt SG, Rizzo J, Sweeney KR, Cropp G, Baker SD, Kraynak MA, Kuhn JG, Villalona-Calero MA, Hammond L, Weiss G, Thurman A, Smith L, Drengler R, Eckardt JR, Moczygemba J, Hannah AL, Von Hoff DD, Rowinsky EK (1999) Phase I and pharmacologic study of the tyrosine kinase inhibitor SU101 in patients with advanced solid tumors. J Clin Oncol 17:1095–1104PubMed

40.

Sulzbacher I, Birner P, Traxler M, Marberger M, Haitel A (2003) Expression of platelet-derived growth factor-alpha alpha receptor is associated with tumor progression in clear cell renal cell carcinoma. Am J Clin Pathol 120:107–112PubMedCrossRef

41.

Bian XW, Du LL, Shi JQ, Cheng YS, Liu FX (2004) Correlation of bFGF, FGFR1 and VEGF expression with vascularity and malignancy of human astrocytomas. Anal Quant Cytol Histol 22:267–274

42.

Yamada SM, Yamaguchi F, Brown R, Berger MS, Morrison RS (1999) Suppression of glioblastoma cell growth following antisense oligonucleotide-mediated inhibition of fibroblast growth factor receptor expression. Glia 28:66–76PubMedCrossRef

43.

Maxwell PH, Dachs GU, Gleadle JM, Nicholls LG, Harris AL, Stratford IJ, Hankinson O, Pugh CW, Ratcliffe PJ (1997) Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth. Proc Natl Acad Sci 94:8104–8109PubMedCrossRef

44.

Thews O, Wolloscheck T, Dillenburg W, Kraus S, Kelleher DK, Konerding MA, Vaupel P (2004) Microenvironmental adaptation of experimental tumors to chronic vs acute hypoxia. Br J cancer 13:1181–1189

45.

Marxsen JH, Schmitt O, Metzen E, Jelkmann W, Hellwig-Burgel T (2001) Vascular endothelial growth factor gene expression in the human breast cancer cell line MX-1 is controlled by the O2 availability in vitro and in vivo. Ann Anat 183:243–249PubMedCrossRef

46.

Yamaguchi KT, Stewart RJ, Wang HM, Hudson SE, Vierra M, Akhtar A et al (1992) Free Radic Res Commun 16:167–174PubMed

47.

Narkowicz CH, Vial JH, McCartney P (1993) Hyperbaric oxygen therapy increases free radical levels in the blood of humans. Free Radical Res Comm 19:71–80

48.

Halliwell B, Gutteridge JM (1990) Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol 186:1–85PubMedCrossRef

49.

Das UN (2002) A radiacl approach to cancer. Med Sci Monit 8:79–92

50.

Conconi MT (2003) Effects of hyperbaric oxygen on proliferative and apoptotic activities and reactive oxygen species generation in mouse fibroblast 3T3/J2 cell line. J Invest Med 51:227–232CrossRef

Title: Hyperoxia retards growth and induces apoptosis, changes in vascular density and gene expression in transplanted gliomas in nude rats
Authors: Linda Elin Birkhaug Stuhr
A. Raa
A. M. Øyan
K. H. Kalland
P. O. Sakariassen
K. Petersen
R. Bjerkvig
R. K. Reed
Publication date: 01-11-2007
Publisher: Springer US
Published in: Journal of Neuro-Oncology / Issue 2/2007
Print ISSN: 0167-594X
Electronic ISSN: 1573-7373
DOI: https://doi.org/10.1007/s11060-007-9407-2

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