The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression
Tumor neo-angiogenesis is regulated, in part, by the hypoxia-inducible gene <em>HIF1</em>.. Evidence suggests <em>HIF1</em> associates with polymerized microtubules and traffics to the nucleus. This study investigated the role of HIF1 in mediating the antitumor activity of tw...
| Main Authors: | , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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International Institute of Anticancer Research
2015
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| _version_ | 1797080311896473600 |
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| author | Stengel, C Newman, S Leese, M Thomas, M Potter, B Reed, M Purohit, A Foster, P |
| author_facet | Stengel, C Newman, S Leese, M Thomas, M Potter, B Reed, M Purohit, A Foster, P |
| author_sort | Stengel, C |
| collection | OXFORD |
| description | Tumor neo-angiogenesis is regulated, in part, by the hypoxia-inducible gene <em>HIF1</em>.. Evidence suggests <em>HIF1</em> associates with polymerized microtubules and traffics to the nucleus. This study investigated the role of HIF1 in mediating the antitumor activity of two steroid-based sulfamate ester microtubule disruptors, STX140 and STX243, <em>in vitro</em> and <em>in vivo</em>. The effects of STX140, STX243 and the parental compound 2-methoxyestradiol (STX66) on HIF1α and HIF2α protein expression were assessed <em>in vitro</em> in MCF-7 and MDA-MB-231 cells cultured under hypoxia. More pertinently, their effects were examined on HIF1-regulated genes <em>in vivo</em> in mice bearing MCF-7 or MDA-MB-231 tumors. The level of mRNA expression of Vascular Endothelial Growth Factor (<em>VEGF</em>), Glucose Transporter 1 (<em>GLUTI</em>), Phosphoglycerate Kinase (<em>PGK</em>), ATP-binding cassette sub-family B member 1 (<em>ABCB1</em>) and Carbonic Anhydrase IX (<em>CAIX</em>) was quantified by Real-time Polymerase Chain Reaction (RT-PCR). Despite inhibiting nuclear HIF1 protein accumulation under hypoxia <em>in vitro</em>, STX140 and STX243 did not significantly regulate the expression of four out of five HIF1α-regulated genes <em>in vitro</em> and <em>in vivo</em>. Only <em>CAIX</em> mRNA expression was down-regulated both <em>in vitro</em> and <em>in vivo</em>. Immunoblot analysis showed that STX140 and STX243 reduced CAIX protein expression <em>in vitro</em>. These compounds had no effect on HIF2α translocation. The potential for inhibition of CAIX by STX140 and STX243 was examined by docking the ligands to the active site in comparison with a known sulfamate-based inhibitor. Microtubule disruption and antitumor activity of STX140 and STX243 is most likely HIF1-independent and may, at least in part, be mediated by inhibition of CAIX expression and activity. |
| first_indexed | 2024-03-07T00:58:12Z |
| format | Journal article |
| id | oxford-uuid:88cfc525-039a-42f0-b8d3-3c0d20fbb4c6 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T00:58:12Z |
| publishDate | 2015 |
| publisher | International Institute of Anticancer Research |
| record_format | dspace |
| spelling | oxford-uuid:88cfc525-039a-42f0-b8d3-3c0d20fbb4c62022-03-26T22:20:03ZThe in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expressionJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:88cfc525-039a-42f0-b8d3-3c0d20fbb4c6EnglishORA DepositInternational Institute of Anticancer Research2015Stengel, CNewman, SLeese, MThomas, MPotter, BReed, MPurohit, AFoster, PTumor neo-angiogenesis is regulated, in part, by the hypoxia-inducible gene <em>HIF1</em>.. Evidence suggests <em>HIF1</em> associates with polymerized microtubules and traffics to the nucleus. This study investigated the role of HIF1 in mediating the antitumor activity of two steroid-based sulfamate ester microtubule disruptors, STX140 and STX243, <em>in vitro</em> and <em>in vivo</em>. The effects of STX140, STX243 and the parental compound 2-methoxyestradiol (STX66) on HIF1α and HIF2α protein expression were assessed <em>in vitro</em> in MCF-7 and MDA-MB-231 cells cultured under hypoxia. More pertinently, their effects were examined on HIF1-regulated genes <em>in vivo</em> in mice bearing MCF-7 or MDA-MB-231 tumors. The level of mRNA expression of Vascular Endothelial Growth Factor (<em>VEGF</em>), Glucose Transporter 1 (<em>GLUTI</em>), Phosphoglycerate Kinase (<em>PGK</em>), ATP-binding cassette sub-family B member 1 (<em>ABCB1</em>) and Carbonic Anhydrase IX (<em>CAIX</em>) was quantified by Real-time Polymerase Chain Reaction (RT-PCR). Despite inhibiting nuclear HIF1 protein accumulation under hypoxia <em>in vitro</em>, STX140 and STX243 did not significantly regulate the expression of four out of five HIF1α-regulated genes <em>in vitro</em> and <em>in vivo</em>. Only <em>CAIX</em> mRNA expression was down-regulated both <em>in vitro</em> and <em>in vivo</em>. Immunoblot analysis showed that STX140 and STX243 reduced CAIX protein expression <em>in vitro</em>. These compounds had no effect on HIF2α translocation. The potential for inhibition of CAIX by STX140 and STX243 was examined by docking the ligands to the active site in comparison with a known sulfamate-based inhibitor. Microtubule disruption and antitumor activity of STX140 and STX243 is most likely HIF1-independent and may, at least in part, be mediated by inhibition of CAIX expression and activity. |
| spellingShingle | Stengel, C Newman, S Leese, M Thomas, M Potter, B Reed, M Purohit, A Foster, P The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title | The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title_full | The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title_fullStr | The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title_full_unstemmed | The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title_short | The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression |
| title_sort | in vitro and in vivo activity of the microtubule disruptor stx140 is mediated by hif 1 alpha and caix expression |
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