Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids

Jesper Kolenda, Stine Skov Jensen, Charlotte Aaberg-Jessen, Karina Christensen, Claus Andersen, Bjarne Winther Kristensen

Publikation: Konferencebidrag uden forlag/tidsskriftPosterForskning

Resumé

Glioblastomas are the most frequent and malignant primary brain tumor. Tumor stem cells in these tumors have recently been suggested to possess innate resistance mechanisms against radiation and chemotherapy possibly explaining their high level of therapeutic resistance. Moreover tumor hypoxia with oxygen tensions below 1-5% O2 has been attributed to play a crucial role in tumorigenesis and therapeutic resistance in glioblastoma. This is in contrast to most in vitro experiments in this field being performed in atmospheric air with 21% O2. In this study the influence of hypoxia on the expression of a panel of stem cell and chemosensitivity markers was therefore investigated using glioma spheroids derived from the conventional glioblastoma cell line U87. The glioma spheroids were derived at normoxic (21% O2) and hypoxic (1% O2) culturing conditions in serum-free medium with EGF and bFGF. The entire immunohistochemical panel included hypoxia (HIF-1α, HIF-2α), proliferation (Ki-67) and stem cell (CD133, nestin, podoplanin, Bmi-1, Sox-2) markers as well as markers related to chemosensitivity (MGMT, MDR-1, TIMP-1, Lamp-1). Since spheroids derived in hypoxia were smaller than in normoxia, a set of experiments was included, in which the culturing time of hypoxic spheroids was extended to obtain equally sized spheroids. Key regulator of the hypoxic response HIF-1α was significantly increased in hypoxia, whereas the Ki-67 positive fraction of proliferative cells was significantly reduced. The expression of stem cell markers CD133, nestin, podoplanin and Bmi-1 was found to be significantly increased in hypoxia similar to what was found for TIMP-1 and Lamp-1. Similar differences in the expression pattern was observed in equally sized normoxic and hypoxic spheroids, although more pronounced differences were found for podoplanin, nestin and TIMP-1 as well as for Ki-67. Hif-2α, Sox-2, MGMT and MDR-1 were not detectable in normoxic and hypoxic U87 spheroids. In conclusion, the expression of tumor stem cell and chemosensitivity markers seems to depend on the oxygen tension suggesting that future development of therapeutic strategies targeting tumors stem cells should take this experimental design into account.
OriginalsprogEngelsk
Publikationsdato2009
StatusUdgivet - 2009
Begivenhed100th Annual Meeting of the American Association for Cancer Research - Denver, USA
Varighed: 18. apr. 200922. apr. 2009

Konference

Konference100th Annual Meeting of the American Association for Cancer Research
LandUSA
ByDenver
Periode18/04/200922/04/2009

Fingeraftryk

Glioblastoma
Nestin
Tissue Inhibitor of Metalloproteinase-1
Cell Line
Glioma
Oxygen
Serum-Free Culture Media
Epidermal Growth Factor
Research Design
Neoplasms

Bibliografisk note

Titel på proceedings: Proceedings of the 100th Annual Meeting of the American Association for Cancer Research
Volume: 100

Citer dette

Kolenda, J., Jensen, S. S., Aaberg-Jessen, C., Christensen, K., Andersen, C., & Kristensen, B. W. (2009). Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids. Poster session præsenteret på 100th Annual Meeting of the American Association for Cancer Research, Denver, USA.
Kolenda, Jesper ; Jensen, Stine Skov ; Aaberg-Jessen, Charlotte ; Christensen, Karina ; Andersen, Claus ; Kristensen, Bjarne Winther. / Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids. Poster session præsenteret på 100th Annual Meeting of the American Association for Cancer Research, Denver, USA.
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title = "Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids",
abstract = "Glioblastomas are the most frequent and malignant primary brain tumor. Tumor stem cells in these tumors have recently been suggested to possess innate resistance mechanisms against radiation and chemotherapy possibly explaining their high level of therapeutic resistance. Moreover tumor hypoxia with oxygen tensions below 1-5{\%} O2 has been attributed to play a crucial role in tumorigenesis and therapeutic resistance in glioblastoma. This is in contrast to most in vitro experiments in this field being performed in atmospheric air with 21{\%} O2. In this study the influence of hypoxia on the expression of a panel of stem cell and chemosensitivity markers was therefore investigated using glioma spheroids derived from the conventional glioblastoma cell line U87. The glioma spheroids were derived at normoxic (21{\%} O2) and hypoxic (1{\%} O2) culturing conditions in serum-free medium with EGF and bFGF. The entire immunohistochemical panel included hypoxia (HIF-1α, HIF-2α), proliferation (Ki-67) and stem cell (CD133, nestin, podoplanin, Bmi-1, Sox-2) markers as well as markers related to chemosensitivity (MGMT, MDR-1, TIMP-1, Lamp-1). Since spheroids derived in hypoxia were smaller than in normoxia, a set of experiments was included, in which the culturing time of hypoxic spheroids was extended to obtain equally sized spheroids. Key regulator of the hypoxic response HIF-1α was significantly increased in hypoxia, whereas the Ki-67 positive fraction of proliferative cells was significantly reduced. The expression of stem cell markers CD133, nestin, podoplanin and Bmi-1 was found to be significantly increased in hypoxia similar to what was found for TIMP-1 and Lamp-1. Similar differences in the expression pattern was observed in equally sized normoxic and hypoxic spheroids, although more pronounced differences were found for podoplanin, nestin and TIMP-1 as well as for Ki-67. Hif-2α, Sox-2, MGMT and MDR-1 were not detectable in normoxic and hypoxic U87 spheroids. In conclusion, the expression of tumor stem cell and chemosensitivity markers seems to depend on the oxygen tension suggesting that future development of therapeutic strategies targeting tumors stem cells should take this experimental design into account.",
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note = "Titel p{\aa} proceedings: Proceedings of the 100th Annual Meeting of the American Association for Cancer Research Volume: 100; 100th Annual Meeting of the American Association for Cancer Research ; Conference date: 18-04-2009 Through 22-04-2009",
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Kolenda, J, Jensen, SS, Aaberg-Jessen, C, Christensen, K, Andersen, C & Kristensen, BW 2009, 'Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids', 100th Annual Meeting of the American Association for Cancer Research, Denver, USA, 18/04/2009 - 22/04/2009.

Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids. / Kolenda, Jesper; Jensen, Stine Skov; Aaberg-Jessen, Charlotte; Christensen, Karina; Andersen, Claus; Kristensen, Bjarne Winther.

2009. Poster session præsenteret på 100th Annual Meeting of the American Association for Cancer Research, Denver, USA.

Publikation: Konferencebidrag uden forlag/tidsskriftPosterForskning

TY - CONF

T1 - Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids

AU - Kolenda, Jesper

AU - Jensen, Stine Skov

AU - Aaberg-Jessen, Charlotte

AU - Christensen, Karina

AU - Andersen, Claus

AU - Kristensen, Bjarne Winther

N1 - Titel på proceedings: Proceedings of the 100th Annual Meeting of the American Association for Cancer Research Volume: 100

PY - 2009

Y1 - 2009

N2 - Glioblastomas are the most frequent and malignant primary brain tumor. Tumor stem cells in these tumors have recently been suggested to possess innate resistance mechanisms against radiation and chemotherapy possibly explaining their high level of therapeutic resistance. Moreover tumor hypoxia with oxygen tensions below 1-5% O2 has been attributed to play a crucial role in tumorigenesis and therapeutic resistance in glioblastoma. This is in contrast to most in vitro experiments in this field being performed in atmospheric air with 21% O2. In this study the influence of hypoxia on the expression of a panel of stem cell and chemosensitivity markers was therefore investigated using glioma spheroids derived from the conventional glioblastoma cell line U87. The glioma spheroids were derived at normoxic (21% O2) and hypoxic (1% O2) culturing conditions in serum-free medium with EGF and bFGF. The entire immunohistochemical panel included hypoxia (HIF-1α, HIF-2α), proliferation (Ki-67) and stem cell (CD133, nestin, podoplanin, Bmi-1, Sox-2) markers as well as markers related to chemosensitivity (MGMT, MDR-1, TIMP-1, Lamp-1). Since spheroids derived in hypoxia were smaller than in normoxia, a set of experiments was included, in which the culturing time of hypoxic spheroids was extended to obtain equally sized spheroids. Key regulator of the hypoxic response HIF-1α was significantly increased in hypoxia, whereas the Ki-67 positive fraction of proliferative cells was significantly reduced. The expression of stem cell markers CD133, nestin, podoplanin and Bmi-1 was found to be significantly increased in hypoxia similar to what was found for TIMP-1 and Lamp-1. Similar differences in the expression pattern was observed in equally sized normoxic and hypoxic spheroids, although more pronounced differences were found for podoplanin, nestin and TIMP-1 as well as for Ki-67. Hif-2α, Sox-2, MGMT and MDR-1 were not detectable in normoxic and hypoxic U87 spheroids. In conclusion, the expression of tumor stem cell and chemosensitivity markers seems to depend on the oxygen tension suggesting that future development of therapeutic strategies targeting tumors stem cells should take this experimental design into account.

AB - Glioblastomas are the most frequent and malignant primary brain tumor. Tumor stem cells in these tumors have recently been suggested to possess innate resistance mechanisms against radiation and chemotherapy possibly explaining their high level of therapeutic resistance. Moreover tumor hypoxia with oxygen tensions below 1-5% O2 has been attributed to play a crucial role in tumorigenesis and therapeutic resistance in glioblastoma. This is in contrast to most in vitro experiments in this field being performed in atmospheric air with 21% O2. In this study the influence of hypoxia on the expression of a panel of stem cell and chemosensitivity markers was therefore investigated using glioma spheroids derived from the conventional glioblastoma cell line U87. The glioma spheroids were derived at normoxic (21% O2) and hypoxic (1% O2) culturing conditions in serum-free medium with EGF and bFGF. The entire immunohistochemical panel included hypoxia (HIF-1α, HIF-2α), proliferation (Ki-67) and stem cell (CD133, nestin, podoplanin, Bmi-1, Sox-2) markers as well as markers related to chemosensitivity (MGMT, MDR-1, TIMP-1, Lamp-1). Since spheroids derived in hypoxia were smaller than in normoxia, a set of experiments was included, in which the culturing time of hypoxic spheroids was extended to obtain equally sized spheroids. Key regulator of the hypoxic response HIF-1α was significantly increased in hypoxia, whereas the Ki-67 positive fraction of proliferative cells was significantly reduced. The expression of stem cell markers CD133, nestin, podoplanin and Bmi-1 was found to be significantly increased in hypoxia similar to what was found for TIMP-1 and Lamp-1. Similar differences in the expression pattern was observed in equally sized normoxic and hypoxic spheroids, although more pronounced differences were found for podoplanin, nestin and TIMP-1 as well as for Ki-67. Hif-2α, Sox-2, MGMT and MDR-1 were not detectable in normoxic and hypoxic U87 spheroids. In conclusion, the expression of tumor stem cell and chemosensitivity markers seems to depend on the oxygen tension suggesting that future development of therapeutic strategies targeting tumors stem cells should take this experimental design into account.

KW - Glioma

KW - Hypoxia

KW - Tumor stem cells

KW - Immunohistochemistry

KW - CD133

M3 - Poster

ER -

Kolenda J, Jensen SS, Aaberg-Jessen C, Christensen K, Andersen C, Kristensen BW. Effects of hypoxia on expression of a panel of stem cell and chemosensitivity markers in glioblastoma cell line-derived spheroids. 2009. Poster session præsenteret på 100th Annual Meeting of the American Association for Cancer Research, Denver, USA.