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Anti-Apoptotic Action by Hypoxia Inducible Factor 1-Alpha in Human Pituitary Adenoma Cell Line, HP-75 in Hypoxic Condition

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Summary

Hypoxia-inducible factor-1 (HIF-1) alpha is the major transcription factor involved in the adaptive response to hypoxia. The purpose of this study was to investigate whether HIF 1-alpha protects HP75 cells, pituitary adenoma cell line from hypoxia induced apoptosis. HP75 was transfected with siRNA targeting HIF 1-alpha mRNA sequences or scrambled RNA duplexes, followed by subjected to hypoxia (1% oxygen) for 24 h, compared with normoxia (21%). The efficacy of RNAi was assessed via real-time RT-PCR and immunohistochemistry. Apoptosis was determined by Tdt-mediated dUTP nick end-labeling (TUNEL) assay and agarose gel electrophoresis. Membrane cDNA microarray was examined to detect gene profiling among the cell in normoxia, hypoxia, or hypoxia following the RNAi. A significantly greater proportion of HP75 cells transfected with specific siRNA duplexes and subsequently exposed to hypoxia demonstrated apoptosis to a large extent when compared with non-transfected cells. Transfection with specific siRNA duplexes knocked down HIF 1-alpha mRNA and protein expression in hypoxia-exposed cells by approximately 80%, whereas transfection with scrambled siRNA duplexes had no noticeable effect on HIF 1-alpha expression. Microarray analysis indicated that HIF1-alpha down-regulated caspase-10. These findings strongly suggest that HIF 1-alpha exerts an antiapoptotic role in HP75 in hypoxia.

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References

  1. Tang K, Breen EC, Wagner H, Brutsaert TD, Gassmann M, Wagner PD, HIF and VEGF relationships in response to hypoxia and sciatic nerve stimulation in rat gastrocnemius Respir Physiol Neurobiol 144: 71–80, 2004

    Article  PubMed  CAS  Google Scholar 

  2. Vidal S, Horvath E, Kovacs K, Kuroki T, Lloyd RV, Scheithauer BW, Expression of hypoxia-inducible factor-1alpha (HIF-1alpha) in pituitary tumours Histol Histopathol 18: 679–686, 2003

    PubMed  CAS  Google Scholar 

  3. DeHahn KC, Gonzales M, Gonzalez AM, Hopkinson SB, Chandel NS, Brunelle JK, Jones JC, The alpha4 laminin subunit regulates endothelial cell survival Exp Cell Res 294: 281–289, 2004

    Article  PubMed  CAS  Google Scholar 

  4. Hammar E, Parnaud G, Bosco D, Perriraz N, Maedler K, Donath M, Rouiller DG, Halban PA, Extracellular matrix protects pancreatic beta-cells against apoptosis: role of short- and long-term signaling pathways Diabetes 53: 2034–2041, 2004

    Article  PubMed  CAS  Google Scholar 

  5. Sarkar D, Boukerche H, Su ZZ, Fisher PB, mda-9/syntenin: recent insights into a novel cell signaling and metastasis-associated gene Pharmacol Ther 104: 101–115, 2004

    Article  PubMed  CAS  Google Scholar 

  6. Kothari S, Cizeau J, McMillan-Ward E, Israels SJ, Bailes M, Ens K, Kirshenbaum LA, Gibson SB, BNIP3 plays a role in hypoxic cell death in human epithelial cells that is inhibited by growth factors EGF and IGF Oncogene 22: 4734–4744, 2003

    Article  PubMed  CAS  Google Scholar 

  7. Baek JH, Jang JE, Kang CM, Chung HY, Kim ND, Kim KW, Hypoxia-induced VEGF enhances tumor survivability via suppression of serum deprivation-induced apoptosis Oncogene 19: 4621–4631, 2000

    Article  PubMed  CAS  Google Scholar 

  8. Shannon AM, Bouchier-Hayes DJ, Condron CM, Toomey D, Tumour hypoxia, chemotherapeutic resistance and hypoxia-related therapies Cancer Treat Rev 29: 297–307, 2003

    Article  PubMed  CAS  Google Scholar 

  9. Goda N, Dozier SJ, Johnson RS, HIF-1 in cell cycle regulation, apoptosis, and tumor progression Antioxid Redox Signal 5: 467–473, 2003

    Article  PubMed  CAS  Google Scholar 

  10. Cerman J, Cap J, Marekova M, Nemecek S, Marek J, Rudolf E, Cervinka M, The role of apoptosis in pituitary adenomas in the field of conventionally used therapeutic approaches Ann N Y Acad Sci 1010: 520–524, 2003

    Article  PubMed  CAS  Google Scholar 

  11. Kristof RA, Aliashkevich AF, Hans V, Haun D, Meyer B, Thees C, Schramm J: The regional oxygen saturation of pituitary adenomas is lower than that of the pituitary gland: microspectrophotometric study with potential clinical implications Neurosurgery 53: 880–885; discussion 885–886, 2003

  12. Iida T, Mine S, Fujimoto H, Suzuki K, Minami Y, Tanaka Y, Hypoxia-inducible factor-1alpha induces cell cycle arrest of endothelial cells Genes Cells 7: 143–149, 2002

    Article  PubMed  CAS  Google Scholar 

  13. Kapranos N, Kontogeorgos G, Horvath E, Kovacs K, Morphology, molecular regulation and significance of apoptosis in pituitary adenomas Front Horm Res 32: 217–234, 2004

    PubMed  CAS  Google Scholar 

  14. Li J, Zhang X, Sejas DP, Bagby GC, Pang Q, Hypoxia-induced nucleophosmin protects cell death through inhibition of p53 J Biol Chem 279: 41275–41279, 2004

    Article  PubMed  CAS  Google Scholar 

  15. Maran J, Prchal J, Polycythemia and oxygen sensing Pathol Biol (Paris) 52: 280–284, 2004

    CAS  Google Scholar 

  16. Meissner U, Allabauer I, Repp R, Rascher W, Dotsch J, Inducible expression of hypoxia-inducible factor 1alpha (HIF-1alpha) as a tool for studying HIF-1alpha-dependent gene regulation during normoxia in vitro Pharmacology 69: 74–78, 2003

    Article  PubMed  CAS  Google Scholar 

  17. Minet E, Michel G, Remacle J, Michiels C, Role of HIF-1 as a transcription factor involved in embryonic development, cancer progression and apoptosis (review) Int J Mol Med 5: 253–259, 2000

    PubMed  CAS  Google Scholar 

  18. Mizutani A, Furukawa T, Adachi Y, Ikehara S, Taketani S, A zinc-finger protein, PLAGL2, induces the expression of a proapoptotic protein Nip3, leading to cellular apoptosis J Biol Chem 277: 15851–15858, 2002

    Article  PubMed  CAS  Google Scholar 

  19. Moeller BJ, Cao Y, Li CY, Dewhirst MW, Radiation activates HIF-1 to regulate vascular radiosensitivity in tumors: role of reoxygenation, free radicals, and stress granules Cancer Cell 5: 429–441, 2004

    Article  PubMed  CAS  Google Scholar 

  20. Moeller BJ, Dewhirst MW, Raising the Bar: How HIF-1 Helps Determine Tumor Radiosensitivity. Cell Cycle 3: 2004

  21. Moley KH, Mueckler MM, Glucose transport and apoptosis Apoptosis 5: 99–105, 2000

    Article  PubMed  CAS  Google Scholar 

  22. Mooberry SL, Mechanism of action of 2-methoxyestradiol: new developments Drug Resist Updat 6: 355–361, 2003

    Article  PubMed  CAS  Google Scholar 

  23. Moritz W, Meier F, Stroka DM, Giuliani M, Kugelmeier P, Nett PC, Lehmann R, Candinas D, Gassmann M, Weber M, Apoptosis in hypoxic human pancreatic islets correlates with HIF-1alpha expression Faseb J 16: 745–747, 2002

    PubMed  CAS  Google Scholar 

  24. Pan Y, Oprysko PR, Asham AM, Koch CJ, Simon MC, p53 cannot be induced by hypoxia alone but responds to the hypoxic microenvironment Oncogene 23: 4975–4983, 2004

    Article  PubMed  CAS  Google Scholar 

  25. Dai S, Huang ML, Hsu CY, Chao KS, Inhibition of hypoxia inducible factor 1alpha causes oxygen-independent cytotoxicity and induces p53 independent apoptosis in glioblastoma cells Int J Radiat Oncol Biol Phys 55: 1027–1036, 2003

    PubMed  CAS  Google Scholar 

  26. Giatromanolaki A, Sivridis E, Maltezos E, Papazoglou D, Simopoulos C, Gatter KC, Harris AL, Koukourakis MI, Hypoxia inducible factor 1alpha and 2alpha overexpression in inflammatory bowel disease J Clin Pathol 56: 209–213, 2003

    Article  PubMed  CAS  Google Scholar 

  27. Koshiji M, Kageyama Y, Pete EA, Horikawa I, Barrett JC, Huang LE, HIF-1alpha induces cell cycle arrest by functionally counteracting Myc Embo J 23: 1949–1956, 2004

    Article  CAS  Google Scholar 

  28. Kotch LE, Iyer NV, Laughner E, Semenza GL, Defective vascularization of HIF-1alpha-null embryos is not associated with VEGF deficiency but with mesenchymal cell death Dev Biol 209: 254–267, 1999

    Article  PubMed  CAS  Google Scholar 

  29. Achison M, Hupp TR, Hypoxia attenuates the p53 response to cellular damage Oncogene 22: 3431–3440, 2003

    Article  PubMed  CAS  Google Scholar 

  30. Harris AL, Hypoxia–a key regulatory factor in tumour growth Nat Rev Cancer 2: 38–47, 2002

    Article  PubMed  CAS  Google Scholar 

  31. Ibrahim AE, Pickering RM, Gawne-Cain ML, King S, Lees PD, Ellison DW, Indices of apoptosis and proliferation as potential prognostic markers in non-functioning pituitary adenomas Clin Neuropathol 23: 8–15, 2004

    PubMed  CAS  Google Scholar 

  32. Akakura N, Kobayashi M, Horiuchi I, Suzuki A, Wang J, Chen J, Niizeki H, Kawamura K, Hosokawa M, Asaka M, Constitutive expression of hypoxia-inducible factor-1alpha renders pancreatic cancer cells resistant to apoptosis induced by hypoxia and nutrient deprivation Cancer Res 61: 6548–6554, 2001

    PubMed  CAS  Google Scholar 

  33. Greijer AE, van der Wall E, The role of hypoxia inducible factor 1 (HIF-1) in hypoxia induced apoptosis J Clin Pathol 57: 1009–1014, 2004

    Article  PubMed  CAS  Google Scholar 

  34. Yu EZ, Li YY, Liu XH, Kagan E, McCarron RM, Antiapoptotic action of hypoxia-inducible factor-1 alpha in human endothelial cells Lab Invest 84: 553–561, 2004

    Article  PubMed  CAS  Google Scholar 

  35. Zhang Q, Zhang ZF, Rao JY, Sato JD, Brown J, Messadi DV, Le AD, Treatment with siRNA and antisense oligonucleotides targeted to HIF-1alpha induced apoptosis in human tongue squamous cell carcinomas Int J Cancer 111: 849–857, 2004

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan (No. 14571340 and No. 15390445).

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Authors and Affiliations

  1. Department of Neurosurgery, Nippon Medical School, Tokyo, Japan

    Daizo Yoshida, Kyonsong Kim, Masahiro Noha & Akira Teramoto

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  1. Daizo Yoshida
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  2. Kyonsong Kim
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  3. Masahiro Noha
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  4. Akira Teramoto
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Yoshida, D., Kim, K., Noha, M. et al. Anti-Apoptotic Action by Hypoxia Inducible Factor 1-Alpha in Human Pituitary Adenoma Cell Line, HP-75 in Hypoxic Condition. J Neurooncol 78, 217–225 (2006). https://doi.org/10.1007/s11060-005-9017-9

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  • DOI: https://doi.org/10.1007/s11060-005-9017-9

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