The mRNA-binding protein HuR promotes hypoxia-induced chemoresistance through posttranscriptional regulation of the proto-oncogene PIM1 in pancreatic cancer cells

F. F. Blanco, M. Jimbo, J. Wulfkuhle, I. Gallagher, J. Deng, L. Enyenihi, N. Meisner-Kober, E. Londin, I. Rigoutsos, J. A. Sawicki, M. V. Risbud, A. K. Witkiewicz, P. A. McCue, W. Jiang, H. Rui, C. J. Yeo, E. Petricoin, J. M. Winter, J. R. Brody

Research output: Contribution to journalArticle

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Abstract

Previously, it has been shown that pancreatic ductal adenocarcinoma (PDA) tumors exhibit high levels of hypoxia, characterized by low oxygen pressure (pO2) and decreased O2 intracellular perfusion. Chronic hypoxia is strongly associated with resistance to cytotoxic chemotherapy and chemoradiation in an understudied phenomenon known as hypoxia-induced chemoresistance. The hypoxia-inducible, pro-oncogenic, serine-threonine kinase PIM1 (Proviral Integration site for Moloney murine leukemia virus 1) has emerged as a key regulator of hypoxia-induced chemoresistance in PDA and other cancers. Although its role in therapeutic resistance has been described previously, the molecular mechanism behind PIM1 overexpression in PDA is unknown. Here, we demonstrate that cis-acting AU-rich elements (ARE) present within a 38-base pair region of the PIM1 mRNA 3′-untranslated region mediate a regulatory interaction with the mRNA stability factor HuR (Hu antigen R) in the context of tumor hypoxia. Predominantly expressed in the nucleus in PDA cells, HuR translocates to the cytoplasm in response to hypoxic stress and stabilizes the PIM1 mRNA transcript, resulting in PIM1 protein overexpression. A reverse-phase protein array revealed that HuR-mediated regulation of PIM1 protects cells from hypoxic stress through phosphorylation and inactivation of the apoptotic effector BAD and activation of MEK1/2. Importantly, pharmacological inhibition of HuR by MS-444 inhibits HuR homodimerization and its cytoplasmic translocation, abrogates hypoxia-induced PIM1 overexpression and markedly enhances PDA cell sensitivity to oxaliplatin and 5-fluorouracil under physiologic low oxygen conditions. Taken together, these results support the notion that HuR has prosurvival properties in PDA cells by enabling them with growth advantages in stressful tumor microenvironment niches. Accordingly, these studies provide evidence that therapeutic disruption of HuR's regulation of PIM1 may be a key strategy in breaking an elusive chemotherapeutic resistance mechanism acquired by PDA cells that reside in hypoxic PDA microenvironments.

Original languageEnglish (US)
Pages (from-to)2529-2541
Number of pages13
JournalOncogene
Volume35
Issue number19
DOIs
StatePublished - May 1 2016

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ELAV Proteins
Proto-Oncogenes
Pancreatic Neoplasms
Carrier Proteins
Adenocarcinoma
Messenger RNA
oxaliplatin
AU Rich Elements
Oxygen
Moloney murine leukemia virus
Protein Array Analysis
Tumor Microenvironment
Hypoxia
Protein-Serine-Threonine Kinases
RNA Stability
3' Untranslated Regions
Base Pairing
Fluorouracil
Neoplasms
Cytoplasm

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

The mRNA-binding protein HuR promotes hypoxia-induced chemoresistance through posttranscriptional regulation of the proto-oncogene PIM1 in pancreatic cancer cells. / Blanco, F. F.; Jimbo, M.; Wulfkuhle, J.; Gallagher, I.; Deng, J.; Enyenihi, L.; Meisner-Kober, N.; Londin, E.; Rigoutsos, I.; Sawicki, J. A.; Risbud, M. V.; Witkiewicz, A. K.; McCue, P. A.; Jiang, W.; Rui, H.; Yeo, C. J.; Petricoin, E.; Winter, J. M.; Brody, J. R.

In: Oncogene, Vol. 35, No. 19, 01.05.2016, p. 2529-2541.

Research output: Contribution to journalArticle

Blanco, FF, Jimbo, M, Wulfkuhle, J, Gallagher, I, Deng, J, Enyenihi, L, Meisner-Kober, N, Londin, E, Rigoutsos, I, Sawicki, JA, Risbud, MV, Witkiewicz, AK, McCue, PA, Jiang, W, Rui, H, Yeo, CJ, Petricoin, E, Winter, JM & Brody, JR 2016, 'The mRNA-binding protein HuR promotes hypoxia-induced chemoresistance through posttranscriptional regulation of the proto-oncogene PIM1 in pancreatic cancer cells', Oncogene, vol. 35, no. 19, pp. 2529-2541. https://doi.org/10.1038/onc.2015.325
Blanco, F. F. ; Jimbo, M. ; Wulfkuhle, J. ; Gallagher, I. ; Deng, J. ; Enyenihi, L. ; Meisner-Kober, N. ; Londin, E. ; Rigoutsos, I. ; Sawicki, J. A. ; Risbud, M. V. ; Witkiewicz, A. K. ; McCue, P. A. ; Jiang, W. ; Rui, H. ; Yeo, C. J. ; Petricoin, E. ; Winter, J. M. ; Brody, J. R. / The mRNA-binding protein HuR promotes hypoxia-induced chemoresistance through posttranscriptional regulation of the proto-oncogene PIM1 in pancreatic cancer cells. In: Oncogene. 2016 ; Vol. 35, No. 19. pp. 2529-2541.
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AU - Wulfkuhle, J.

AU - Gallagher, I.

AU - Deng, J.

AU - Enyenihi, L.

AU - Meisner-Kober, N.

AU - Londin, E.

AU - Rigoutsos, I.

AU - Sawicki, J. A.

AU - Risbud, M. V.

AU - Witkiewicz, A. K.

AU - McCue, P. A.

AU - Jiang, W.

AU - Rui, H.

AU - Yeo, C. J.

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AU - Winter, J. M.

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N2 - Previously, it has been shown that pancreatic ductal adenocarcinoma (PDA) tumors exhibit high levels of hypoxia, characterized by low oxygen pressure (pO2) and decreased O2 intracellular perfusion. Chronic hypoxia is strongly associated with resistance to cytotoxic chemotherapy and chemoradiation in an understudied phenomenon known as hypoxia-induced chemoresistance. The hypoxia-inducible, pro-oncogenic, serine-threonine kinase PIM1 (Proviral Integration site for Moloney murine leukemia virus 1) has emerged as a key regulator of hypoxia-induced chemoresistance in PDA and other cancers. Although its role in therapeutic resistance has been described previously, the molecular mechanism behind PIM1 overexpression in PDA is unknown. Here, we demonstrate that cis-acting AU-rich elements (ARE) present within a 38-base pair region of the PIM1 mRNA 3′-untranslated region mediate a regulatory interaction with the mRNA stability factor HuR (Hu antigen R) in the context of tumor hypoxia. Predominantly expressed in the nucleus in PDA cells, HuR translocates to the cytoplasm in response to hypoxic stress and stabilizes the PIM1 mRNA transcript, resulting in PIM1 protein overexpression. A reverse-phase protein array revealed that HuR-mediated regulation of PIM1 protects cells from hypoxic stress through phosphorylation and inactivation of the apoptotic effector BAD and activation of MEK1/2. Importantly, pharmacological inhibition of HuR by MS-444 inhibits HuR homodimerization and its cytoplasmic translocation, abrogates hypoxia-induced PIM1 overexpression and markedly enhances PDA cell sensitivity to oxaliplatin and 5-fluorouracil under physiologic low oxygen conditions. Taken together, these results support the notion that HuR has prosurvival properties in PDA cells by enabling them with growth advantages in stressful tumor microenvironment niches. Accordingly, these studies provide evidence that therapeutic disruption of HuR's regulation of PIM1 may be a key strategy in breaking an elusive chemotherapeutic resistance mechanism acquired by PDA cells that reside in hypoxic PDA microenvironments.

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