Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells

Rekha Rao, Srilatha Nalluri, Ravindra Bharat Kolhe, Yonghua Yang, Warren Fiskus, Jianguang Chen, Kyungsoo Ha, Kathleen M. Buckley, Ramesh Balusu, Veena Coothankandaswamy, Atul Joshi, Peter Atadja, Kapil N. Bhalla

Research output: Contribution to journalArticle

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Abstract

Increased levels of misfolded polypeptides in the endoplasmic reticulum (ER) triggers the dissociation of glucose-regulated protein 78 (GRP78) from the three transmembrane ER-stress mediators, i.e., protein kinase RNA-like ER kinase (PERK), activating transcription factor-6 (ATF6), and inositol-requiring enzyme lα which results in the adaptive unfolded protein response (UPR). In the present studies, we determined that histone deacetylase-6 (HDAC6) binds and deacetylates GRP78. Following treatment with the pan-histone deacetylase inhibitor panobinostat (Novartis Pharmaceuticals), or knockdown of HDAC6 by short hairpin RNA, GRP78 is acetylated in 11 lysine residues, which dissociates GRP78 from PERK. This is associated with the activation of a lethal UPR in human breast cancer cells. Coimmunoprecipitation studies showed that binding of HDAC6 to GRP78 requires the second catalytic and COOH-terminal BUZ domains of HDAC6. Treatment with panobinostat increased the levels of phosphorylated-eukaryotic translation initiation factor (p-eIF2α), ATF4, and CAAT/enhancer binding protein homologous protein (CHOP). Panobinostat treatment also increased the proapoptotic BIK, BIM, BAX, and BAK levels, as well as increased the activity of caspase-7. Knockdown of GRP78 sensitized MCF-7 cells to bortezomib and panobinostat-induced UPR and cell death. These findings indicate that enforced acetylation and decreased binding of GRP78 to PERK is mechanistically linked to panobinostat-induced UPR and cell death of breast cancer cells.

Original languageEnglish (US)
Pages (from-to)942-952
Number of pages11
JournalMolecular cancer therapeutics
Volume9
Issue number4
DOIs
StatePublished - Apr 1 2010

Fingerprint

Endoplasmic Reticulum Stress
Acetylation
Unfolded Protein Response
Breast Neoplasms
Histone Deacetylases
Endoplasmic Reticulum
Activating Transcription Factor 6
Cell Death
CCAAT-Enhancer-Binding Proteins
Eukaryotic Initiation Factors
Caspase 7
Histone Deacetylase Inhibitors
MCF-7 Cells
Inositol
glucose-regulated proteins
panobinostat
Protein Kinases
Small Interfering RNA
Lysine
RNA

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells. / Rao, Rekha; Nalluri, Srilatha; Kolhe, Ravindra Bharat; Yang, Yonghua; Fiskus, Warren; Chen, Jianguang; Ha, Kyungsoo; Buckley, Kathleen M.; Balusu, Ramesh; Coothankandaswamy, Veena; Joshi, Atul; Atadja, Peter; Bhalla, Kapil N.

In: Molecular cancer therapeutics, Vol. 9, No. 4, 01.04.2010, p. 942-952.

Research output: Contribution to journalArticle

Rao, R, Nalluri, S, Kolhe, RB, Yang, Y, Fiskus, W, Chen, J, Ha, K, Buckley, KM, Balusu, R, Coothankandaswamy, V, Joshi, A, Atadja, P & Bhalla, KN 2010, 'Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells', Molecular cancer therapeutics, vol. 9, no. 4, pp. 942-952. https://doi.org/10.1158/1535-7163.MCT-09-0988
Rao, Rekha ; Nalluri, Srilatha ; Kolhe, Ravindra Bharat ; Yang, Yonghua ; Fiskus, Warren ; Chen, Jianguang ; Ha, Kyungsoo ; Buckley, Kathleen M. ; Balusu, Ramesh ; Coothankandaswamy, Veena ; Joshi, Atul ; Atadja, Peter ; Bhalla, Kapil N. / Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells. In: Molecular cancer therapeutics. 2010 ; Vol. 9, No. 4. pp. 942-952.
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AU - Rao, Rekha

AU - Nalluri, Srilatha

AU - Kolhe, Ravindra Bharat

AU - Yang, Yonghua

AU - Fiskus, Warren

AU - Chen, Jianguang

AU - Ha, Kyungsoo

AU - Buckley, Kathleen M.

AU - Balusu, Ramesh

AU - Coothankandaswamy, Veena

AU - Joshi, Atul

AU - Atadja, Peter

AU - Bhalla, Kapil N.

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AB - Increased levels of misfolded polypeptides in the endoplasmic reticulum (ER) triggers the dissociation of glucose-regulated protein 78 (GRP78) from the three transmembrane ER-stress mediators, i.e., protein kinase RNA-like ER kinase (PERK), activating transcription factor-6 (ATF6), and inositol-requiring enzyme lα which results in the adaptive unfolded protein response (UPR). In the present studies, we determined that histone deacetylase-6 (HDAC6) binds and deacetylates GRP78. Following treatment with the pan-histone deacetylase inhibitor panobinostat (Novartis Pharmaceuticals), or knockdown of HDAC6 by short hairpin RNA, GRP78 is acetylated in 11 lysine residues, which dissociates GRP78 from PERK. This is associated with the activation of a lethal UPR in human breast cancer cells. Coimmunoprecipitation studies showed that binding of HDAC6 to GRP78 requires the second catalytic and COOH-terminal BUZ domains of HDAC6. Treatment with panobinostat increased the levels of phosphorylated-eukaryotic translation initiation factor (p-eIF2α), ATF4, and CAAT/enhancer binding protein homologous protein (CHOP). Panobinostat treatment also increased the proapoptotic BIK, BIM, BAX, and BAK levels, as well as increased the activity of caspase-7. Knockdown of GRP78 sensitized MCF-7 cells to bortezomib and panobinostat-induced UPR and cell death. These findings indicate that enforced acetylation and decreased binding of GRP78 to PERK is mechanistically linked to panobinostat-induced UPR and cell death of breast cancer cells.

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