Hypoxia-inducible factor-2α regulates the expression of TRAIL receptor DR5 in renal cancer cells

S. Mahajan, V. Dammai, T. Hsu, Andrew Kraft

Research output: Contribution to journalArticle

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Abstract

To understand the role of hypoxia-inducible factor (HIF)-2α in regulating sensitivity of renal cancer cells to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis, we transfected wild-type and mutant von Hippel Lindau (VHL) proteins into TRAIL-sensitive, VHL-negative A498 cells. We find that wild-type VHL, but not the VHL mutants S65W and C162F that do not degrade HIF proteins, cause TRAIL resistance. Knock down of the HIF-2α protein by RNA interference (short hairpin RNA) blocked TRAIL-induced apoptosis, decreased the level of TRAIL receptor (DR5) protein and inhibited the transcription of DR5 messenger RNA. By using luciferase constructs containing the upstream region of the DR5 promoter, we demonstrate that HIF-2α stimulates the transcription of the DR5 gene by activating the upstream region between -448 and -1188. Because HIF-2α is thought to exert its effect on gene transcription by interacting with the Max protein partner of Myc in the Myc/Max dimer, small interfering RNAs to Myc were used to lower the levels of this protein. In multiple renal cancer cell lines decreasing the levels of Myc blocked the ability of HIF-2α to stimulate DR5 transcription. PS-341 (VELCADE, bortezomib), a proteasome inhibitor used to treat human cancer, increases the levels of both HIF-2α and c-Myc and elevates the level of DR5 in renal cancer, sensitizing renal cancer cells to TRAIL therapy. Similarly, increasing HIF-2α in prostate and lung cancer cell lines increased the levels of DR5. Thus, in renal cancer cell lines expressing HIF-2α, this protein plays a role in regulating the levels of the TRAIL receptor DR5.

Original languageEnglish (US)
Pages (from-to)1734-1741
Number of pages8
JournalCarcinogenesis
Volume29
Issue number9
DOIs
StatePublished - 2008
Externally publishedYes

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TNF-Related Apoptosis-Inducing Ligand Receptors
Renal Cell Carcinoma
Kidney Neoplasms
TNF-Related Apoptosis-Inducing Ligand
Apoptosis
Cell Line
Small Interfering RNA
Proteins
Proteasome Inhibitors
endothelial PAS domain-containing protein 1
RNA Interference
Luciferases
Genetic Promoter Regions
Genes
Lung Neoplasms
Prostatic Neoplasms
Tumor Necrosis Factor-alpha
Ligands
Messenger RNA

ASJC Scopus subject areas

  • Cancer Research

Cite this

Hypoxia-inducible factor-2α regulates the expression of TRAIL receptor DR5 in renal cancer cells. / Mahajan, S.; Dammai, V.; Hsu, T.; Kraft, Andrew.

In: Carcinogenesis, Vol. 29, No. 9, 2008, p. 1734-1741.

Research output: Contribution to journalArticle

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abstract = "To understand the role of hypoxia-inducible factor (HIF)-2α in regulating sensitivity of renal cancer cells to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis, we transfected wild-type and mutant von Hippel Lindau (VHL) proteins into TRAIL-sensitive, VHL-negative A498 cells. We find that wild-type VHL, but not the VHL mutants S65W and C162F that do not degrade HIF proteins, cause TRAIL resistance. Knock down of the HIF-2α protein by RNA interference (short hairpin RNA) blocked TRAIL-induced apoptosis, decreased the level of TRAIL receptor (DR5) protein and inhibited the transcription of DR5 messenger RNA. By using luciferase constructs containing the upstream region of the DR5 promoter, we demonstrate that HIF-2α stimulates the transcription of the DR5 gene by activating the upstream region between -448 and -1188. Because HIF-2α is thought to exert its effect on gene transcription by interacting with the Max protein partner of Myc in the Myc/Max dimer, small interfering RNAs to Myc were used to lower the levels of this protein. In multiple renal cancer cell lines decreasing the levels of Myc blocked the ability of HIF-2α to stimulate DR5 transcription. PS-341 (VELCADE, bortezomib), a proteasome inhibitor used to treat human cancer, increases the levels of both HIF-2α and c-Myc and elevates the level of DR5 in renal cancer, sensitizing renal cancer cells to TRAIL therapy. Similarly, increasing HIF-2α in prostate and lung cancer cell lines increased the levels of DR5. Thus, in renal cancer cell lines expressing HIF-2α, this protein plays a role in regulating the levels of the TRAIL receptor DR5.",
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AB - To understand the role of hypoxia-inducible factor (HIF)-2α in regulating sensitivity of renal cancer cells to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis, we transfected wild-type and mutant von Hippel Lindau (VHL) proteins into TRAIL-sensitive, VHL-negative A498 cells. We find that wild-type VHL, but not the VHL mutants S65W and C162F that do not degrade HIF proteins, cause TRAIL resistance. Knock down of the HIF-2α protein by RNA interference (short hairpin RNA) blocked TRAIL-induced apoptosis, decreased the level of TRAIL receptor (DR5) protein and inhibited the transcription of DR5 messenger RNA. By using luciferase constructs containing the upstream region of the DR5 promoter, we demonstrate that HIF-2α stimulates the transcription of the DR5 gene by activating the upstream region between -448 and -1188. Because HIF-2α is thought to exert its effect on gene transcription by interacting with the Max protein partner of Myc in the Myc/Max dimer, small interfering RNAs to Myc were used to lower the levels of this protein. In multiple renal cancer cell lines decreasing the levels of Myc blocked the ability of HIF-2α to stimulate DR5 transcription. PS-341 (VELCADE, bortezomib), a proteasome inhibitor used to treat human cancer, increases the levels of both HIF-2α and c-Myc and elevates the level of DR5 in renal cancer, sensitizing renal cancer cells to TRAIL therapy. Similarly, increasing HIF-2α in prostate and lung cancer cell lines increased the levels of DR5. Thus, in renal cancer cell lines expressing HIF-2α, this protein plays a role in regulating the levels of the TRAIL receptor DR5.

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