HDAC5–LSD1 axis regulates antineoplastic effect of natural HDAC inhibitor sulforaphane in human breast cancer cells
Chunyu Cao
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
C.C. and H.W. contributed equally to this work
Search for more papers by this authorHao Wu
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, People's Republic of China
C.C. and H.W. contributed equally to this work
Search for more papers by this authorShauna N. Vasilatos
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorUma Chandran
Department of Biomedical Informatics, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorYe Qin
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorYong Wan
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorSteffi Oesterreich
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorNancy E. Davidson
Fred Hutchinson Cancer Research Center, Seattle Cancer Care Alliance, and University of Washington, Seattle, WA
Search for more papers by this authorCorresponding Author
Yi Huang
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Correspondence to: Yi Huang, M.D., Ph.D., Magee Womens Research Institute, Room 406, 204 Craft Ave, Pittsburgh, PA 15213, USA, Tel.: +1-412-641-3589; E-mail: [email protected]Search for more papers by this authorChunyu Cao
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
C.C. and H.W. contributed equally to this work
Search for more papers by this authorHao Wu
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, People's Republic of China
C.C. and H.W. contributed equally to this work
Search for more papers by this authorShauna N. Vasilatos
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorUma Chandran
Department of Biomedical Informatics, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorYe Qin
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorYong Wan
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorSteffi Oesterreich
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Search for more papers by this authorNancy E. Davidson
Fred Hutchinson Cancer Research Center, Seattle Cancer Care Alliance, and University of Washington, Seattle, WA
Search for more papers by this authorCorresponding Author
Yi Huang
Women's Cancer Research Center, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA
Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA
Correspondence to: Yi Huang, M.D., Ph.D., Magee Womens Research Institute, Room 406, 204 Craft Ave, Pittsburgh, PA 15213, USA, Tel.: +1-412-641-3589; E-mail: [email protected]Search for more papers by this authorConflicts of interest: The authors declare no conflict of interest.
Abstract
Our recent studies have shown that cross-talk between histone deacetylase 5 (HDAC5) and lysine-specific demethylase 1 (LSD1) facilitates breast cancer progression. In this work, we demonstrated that regulatory activity at −356 to −100 bp promoter element plays a critical role in governing HDAC5 transcription. By using DNA affinity precipitation and mass spectrometry, we identified a group of factors that bind to this element. Among these factors, Upstream Transcription Factor 1 (USF1) was shown to play a critical role in controlling HDAC5 transcription. Through screening a panel of epigenetic modifying drugs, we showed that a natural bioactive HDAC inhibitor, sulforaphane, downregulated HDAC5 transcription by blocking USF1 activity. Sulforaphane facilitated LSD1 ubiquitination and degradation in an HDAC5-dependent manner. A comparative microarray analysis demonstrated a genome wide cooperative effect of HDAC5 and LSD1 on cancer-related gene expression. shRNA knockdown and sulforaphane inhibition of HDAC5/LSD1 exhibited similar effects on expression of HDAC5/LSD1 target genes. We also showed that coordinated cross-talk of HDAC5 and LSD1 is essential for the antitumor efficacy of sulforaphane. Combination treatment with sulforaphane and a potent LSD1 inhibitor resulted in synergistic growth inhibition in breast cancer cells, but not in normal breast epithelial cells. Furthermore, combined therapy with sulforaphane and LSD1 inhibitor exhibited superior inhibitory effect on MDA-MB-231 xenograft tumor growth. Taken together, our work demonstrates that HDAC5–LSD1 axis is an effective drug target for breast cancer. Inhibition of HDAC5–LSD1 axis with sulforaphane blocks breast cancer growth and combined treatment with LSD1 inhibitor improves the therapeutic efficacy of sulforaphane.
Abstract
What's new?
The post-translational modification of histones through acetylation serves an important role in the regulation of gene expression. Histone deacetylases (HDACs) are a critical component of this system. This study sheds light on regulatory mechanisms of HDAC5 transcription and shows that sulforaphane, an HDAC inhibitor, suppresses HDAC5 expression through downregulation of upstream transcription factor 1 (USF1). USF1 downregulation in turn destabilizes lysine-specific demethylase 1 (LSD1) protein in breast cancer cells. The findings suggest that targeting the HDAC5–LSD1 axis through combined sulforaphane and LSD1 inhibitor treatment could be an effective approach to enhancing antineoplastic efficacy of epigenetic agents against breast cancer.
Supporting Information
Additional Supporting Information may be found in the online version of this article.
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| ijc31419-sup-0001-suppinfoFigS1.tif1.3 MB | Supporting Information Figure S1 |
| ijc31419-sup-0002-suppinfoFigS2.tif347.6 KB | Supporting Information Figure S2 |
| ijc31419-sup-0003-suppinfoFigS3.tif2 MB | Supporting Information Figure S3 |
| ijc31419-sup-0004-suppinfoFigS4.tif5.3 MB | Supporting Information Figure S4 |
| ijc31419-sup-0005-suppinfoFigS5.tif490.3 KB | Supporting Information Figure S5 |
| ijc31419-sup-0006-suppinfoFigS6.tif1.9 MB | Supporting Information Figure S6 |
| ijc31419-sup-0007-suppinfoFigS7.tif12.8 MB | Supporting Information Figure S7 |
| ijc31419-sup-0008-suppinfoFigLeg.docx15.4 KB | Supporting Information Figure Legends |
| ijc31419-sup-0009-suppinfoTabs.docx21 KB | Supporting Information Tables |
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