A Novel IMP1 Inhibitor, BTYNB, Targets c-Myc and Inhibits Melanoma and Ovarian Cancer Cell Proliferation  Lily Mahapatra, Neal Andruska, Chengjian Mao,

Slides:



Advertisements
Similar presentations
Nogo-p4 Suppresses TrkA Signaling Induced by Low Concentrations of Nerve Growth Factor Through NgR1 in Differentiated PC12 Cells Neurosignals 2016;24:25-39.
Advertisements

Overexpression of CRM1: A Characteristic Feature in a Transformed Phenotype of Lung Carcinogenesis and a Molecular Target for Lung Cancer Adjuvant Therapy 
Volume 11, Issue 4, Pages (April 2007)
MicroRNA-101 Inhibits Growth, Proliferation and Migration and Induces Apoptosis of Breast Cancer Cells by Targeting Sex-Determining Region Y-Box 2 Cell.
Volume 131, Issue 4, Pages (October 2006)
Upregulation of miR-142-3p Improves Drug Sensitivity of Acute Myelogenous Leukemia through Reducing P-Glycoprotein and Repressing Autophagy by Targeting.
Tsai-Der Chuang, Ph.D., Omid Khorram, M.D., Ph.D. 
A Signal Transduction Pathway from TGF-β1 to SKP2 via Akt1 and c-Myc and its Correlation with Progression in Human Melanoma  Xuan Qu, Liangliang Shen,
Integrin αvβ6 Promotes Lung Cancer Proliferation and Metastasis through Upregulation of IL-8–Mediated MAPK/ERK Signaling  Pengwei Yan, Huanfeng Zhu, Li.
Volume 15, Issue 6, Pages (June 2012)
SIRT1 Regulates the Chemoresistance and Invasiveness of Ovarian Carcinoma Cells  David Hamisi Mvunta, Tsutomu Miyamoto, Ryoichi Asaka, Yasushi Yamada,
Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.
A Requirement for ZAK Kinase Activity in Canonical TGF-β Signaling
Cdc42 Inhibits ERK-Mediated Collagenase-1 (MMP-1) Expression in Collagen-Activated Human Keratinocytes  Maryam G. Rohani, Brian K. Pilcher, Peter Chen,
Yongping Shao, Kaitlyn Le, Hanyin Cheng, Andrew E. Aplin 
SDHB deficiency promotes TGFβ-mediated invasion and metastasis of colorectal cancer through transcriptional repression complex SNAIL1-SMAD3/4  Haiyu Wang,
MiR-29 Regulates Type VII Collagen in Recessive Dystrophic Epidermolysis Bullosa  Michael Vanden Oever, Daniel Muldoon, Wendy Mathews, Ron McElmurry, Jakub.
IFN-γ Induces Gastric Cancer Cell Proliferation and Metastasis Through Upregulation of Integrin β3-Mediated NF-κB Signaling  Yuan-Hua Xu, Zheng-Li Li,
MDH2 Stimulated by Estrogen-GPR30 Pathway Down-Regulated PTEN Expression Promoting the Proliferation and Invasion of Cells in Endometrial Cancer  Yan.
Volume 137, Issue 2, Pages e2 (August 2009)
MicroRNA-489 Plays an Anti-Metastatic Role in Human Hepatocellular Carcinoma by Targeting Matrix Metalloproteinase-7  Yixiong Lin, Jianjun Liu, Yuqi Huang,
Characterization of TNF-α– and IL-17A–Mediated Synergistic Induction of DEFB4 Gene Expression in Human Keratinocytes through IκBζ  Claus Johansen, Trine.
Volume 33, Issue 2, Pages (January 2009)
A Requirement for ZAK Kinase Activity in Canonical TGF-β Signaling
MicroRNA-558 regulates the expression of cyclooxygenase-2 and IL-1β-induced catabolic effects in human articular chondrocytes  S.J. Park, E.J. Cheon,
Volume 39, Issue 4, Pages (August 2010)
Epidermal Growth Factor Induces Fibronectin Expression in Human Dermal Fibroblasts via Protein Kinase C δ Signaling Pathway  Yoshihiro Mimura, Hironobu.
Volume 16, Issue 4, Pages (October 2009)
Volume 28, Issue 4, Pages (October 2015)
Volume 22, Issue 12, Pages (December 2015)
Volume 131, Issue 4, Pages (October 2006)
Combining the Multitargeted Tyrosine Kinase Inhibitor Vandetanib with the Antiestrogen Fulvestrant Enhances Its Antitumor Effect in Non-small Cell Lung.
Volume 130, Issue 3, Pages (September 2013)
Andreea M. Bujor, Jaspreet Pannu, Shizhong Bu, Edwin A. Smith, Robin C
Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.
Uc.454 Inhibited Growth by Targeting Heat Shock Protein Family A Member 12B in Non- Small-Cell Lung Cancer  Jun Zhou, Chenghai Wang, Weijuan Gong, Yandan.
SiRNA-Targeting Transforming Growth Factor-β Type I Receptor Reduces Wound Scarring and Extracellular Matrix Deposition of Scar Tissue  Yi-Wen Wang, Nien-Hsien.
Molecular Therapy - Nucleic Acids
A Constitutive Intrinsic Inflammatory Signaling Circuit Composed of miR-196b, Meis2, PPP3CC, and p65 Drives Prostate Cancer Castration Resistance  Ji-Hak.
MicroRNA-101 Exerts Tumor-Suppressive Functions in Non-small Cell Lung Cancer through Directly Targeting Enhancer of Zeste Homolog 2  Ji-guang Zhang,
Inhibition of KLF4 by Statins Reverses Adriamycin-Induced Metastasis and Cancer Stemness in Osteosarcoma Cells  Yangling Li, Miao Xian, Bo Yang, Meidan.
Volume 23, Issue 10, Pages (October 2016)
Volume 129, Issue 5, Pages (November 2005)
The TRAF-Interacting Protein (TRIP) Is a Regulator of Keratinocyte Proliferation  Stéphanie Almeida, Stephan Ryser, Magdalena Obarzanek-Fojt, Daniel Hohl,
Volume 43, Issue 6, Pages (September 2011)
Volume 29, Issue 4, Pages (February 2008)
Volume 132, Issue 5, Pages (May 2007)
Volume 4, Issue 3, Pages (March 2015)
Volume 31, Issue 4, Pages (August 2008)
Histamine Enhances the Production of Granulocyte-Macrophage Colony-Stimulating Factor via Protein Kinase Cα and Extracellular Signal-Regulated Kinase.
miR-124 Inhibits Lung Tumorigenesis Induced by K-ras Mutation and NNK
Bo-Kuan Wu, Charles Brenner  Cell Reports 
Promotion Effects of miR-375 on the Osteogenic Differentiation of Human Adipose- Derived Mesenchymal Stem Cells  Si Chen, Yunfei Zheng, Shan Zhang, Lingfei.
P. Harding, L. Balasubramanian, J. Swegan, A. Stevens, W.F. Glass 
17β-estradiol Inhibits the Production of RANTES in Human Keratinocytes
Volume 20, Issue 4, Pages (October 2011)
Volume 19, Issue 8, Pages (August 2011)
SIRT1, a Class III Histone Deacetylase, Regulates LPS-Induced Inflammation in Human Keratinocytes and Mediates the Anti-Inflammatory Effects of Hinokitiol 
Volume 67, Issue 6, Pages (June 2005)
MELK Promotes Melanoma Growth by Stimulating the NF-κB Pathway
Volume 16, Issue 1, Pages (June 2016)
Volume 70, Issue 5, Pages (September 2006)
Volume 21, Issue 1, Pages (January 2013)
Loss of HDAC-Mediated Repression and Gain of NF-κB Activation Underlie Cytokine Induction in ARID1A- and PIK3CA-Mutation-Driven Ovarian Cancer  Minchul.
Volume 12, Issue 1, Pages (January 2019)
Naoko Kanda, Shinichi Watanabe  Journal of Investigative Dermatology 
Volume 11, Issue 1, Pages (January 2005)
Volume 22, Issue 12, Pages (December 2015)
c-IAP1 Cooperates with Myc by Acting as a Ubiquitin Ligase for Mad1
Overexpression of CRM1: A Characteristic Feature in a Transformed Phenotype of Lung Carcinogenesis and a Molecular Target for Lung Cancer Adjuvant Therapy 
Presentation transcript:

A Novel IMP1 Inhibitor, BTYNB, Targets c-Myc and Inhibits Melanoma and Ovarian Cancer Cell Proliferation  Lily Mahapatra, Neal Andruska, Chengjian Mao, Jeremy Le, David J. Shapiro  Translational Oncology  Volume 10, Issue 5, Pages 818-827 (October 2017) DOI: 10.1016/j.tranon.2017.07.008 Copyright © 2017 The Authors Terms and Conditions

Figure 1 BTYNB is a structurally selective inhibitor of IMP1 binding to flMyc. Dose-response studies of the effect of (A) BTYNB and (B) Compound 5226752 on the binding of IMP1 to a flMyc and binding of purified full-length human PR binding to an flPRE. Compound 5226752 is a BTYNB-related compound. No inhibitor control was set to 100%. Data are mean±SEM (n=5). Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions

Figure 2 BTYNB decreases c-Myc protein levels by decreasing c-Myc mRNA stability, and BTYNB decreases IMP1 protein levels. (A) IMP1 protein levels in 31 cancer cell lines. (B) BYTNB enhances the degradation rate of c-Myc mRNA in SK-MEL2 cells. Cells pretreated with DMSO or 10 μM BTYNB for 72 hours, followed by treatment with actinomycin D for the indicated times. P values testing for significance between the vehicle control (DMSO)- and BTYNB-treated cells. (C) qRT-PCR analysis of c-Myc mRNA levels after treating SK-MEL2 melanoma cells for 72 hours with DMSO or 10 μM BTYNB (n=3; DMSO-vehicle control set to 100%). (D) Western blot analysis of c-Myc protein levels after treating SK-MEL2 cells for 72 hours with increasing concentrations of BTYNB. (E) RNAi knockdown of IMP1 reduces c-Myc mRNA levels in SK-MEL2 cells (n=3; NC siRNA set to 100%). Cells were treated with 100 nM NC or IMP1 siRNA SmartPool, and 72 hours later, c-Myc mRNA levels were quantitated via qRT-PCR. (F) Western blotting analysis of IMP1 protein levels in IGROV-1 cells treated for 72 hours with increasing concentrations of BTYNB. (G) Western blotting analysis of IMP1 protein levels in SK-MEL2 cells treated for 72 hours with increasing concentrations of BTYNB. Data reported as mean±SEM. P values represent comparison to DMSO control (C, E). *P<.05, **P<.01, ***P<.001. Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions

Figure 3 IMP1 stabilizes several mRNA transcripts in melanoma and ovarian cancer cells, which are upregulated in clinical tumor samples. RNAi knockdown of IMP1 decreases the mRNA levels of IMP1-regulated genes in (A) SK-MEL2 melanoma cells and (B) IGROV-1 ovarian cancer cells (n=4; NC set to 100%). Cells were treated with 100 nM NC or IMP1 siRNA SmartPools. (C) NF-κB luciferase activity after treating IGROV-1 cells with 100 nM NC or IMP1 siRNA SmartPool (n=3). (D) Western blot analysis of eEF2 protein levels following treatment of IGROV-1 cells with 100 nM NC or IMP1 siRNA SmartPool. (E) Protein synthesis in IGROV-1 ovarian cancer after treating with 100 nM NC or IMP1 siRNA SmartPool (n=5; NC set to 100%). (F) IMP1 expression in patient samples of normal peritoneum (n=8) or serous adenocarcinoma (n=45). (G) Expression levels of IMP1 and IMP1-regulated genes in samples of normal skin (n=4) and cutaneous melanoma (n=14). Data reported as mean±SEM. P values represent comparison to NC control (A, B, C, E) or histologically normal tissue (F, G). *P<.05, ** P<.01, ***P<.001. ns, not significant. Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions

Figure 4 BTYNB selectively decreases levels of IMP1-regulated mRNAs, decreases protein synthesis, and decreases NF-κB activity. (A) qRT-PCR analysis of IMP1-regulated genes after treating (A) SK-MEL2 melanoma or (B) IGROV-1 ovarian cancer cells with DMSO-vehicle control or 10 μM BTYNB for 72 hours (n=4; DMSO-vehicle control set to 100%). (C) BTYNB inhibits NF-κB luciferase activity in IGROV-1 ovarian cancer cells. Cells were treated with 10 μM BTYNB for 72 hours (n=3). (D) Western blot analysis of eEF2 protein levels in SK-MEL2 cells treated with DMSO-vehicle control or 10 μM BTYNB for 72 hours. (E) Protein synthesis in SK-MEL2 melanoma cancer cells treated with DMSO-vehicle control or 10 μM BTYNB for 72 hours (n=6; DMSO-vehicle control set to 100%). (F) BTYNB does not inhibit ERα or GR activity in T47D-kBluc or T47D (A1-2) cells, respectively (n=6; DMSO-vehicle control set to 100%). Cells were treated for 24 hours in the presence or absence of 10 nM 17β-estradiol in T47D-KBluc cells or 10 nM dexamethasone in T47D/(A1-2) cells. Cells treated with hormone were treated with the indicated concentrations of BTYNB (or DMSO-vehicle control). P values testing for significance between the vehicle-control (DMSO) and BTYNB-treated cells in the presence of hormone. Data reported as mean±SEM. P values represent comparison to DMSO control (A, B, C, E). *P<.05, **P<.01, ***P<.001. ns, not significant. Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions

Figure 5 IMP1 depletion and BTYNB block proliferation of IMP1-positive cancer cells, and overexpression of IMP1 is sufficient to reverse BTYNB-mediated inhibition of cell proliferation. siRNA knockdown of IMP1 blocks proliferation of IMP1-positive (A) SK-MEL2 melanoma, (B) IGROV-1 ovarian, and (C) ES-2 ovarian cancer cells with no effects on IMP1-negative (D) BG-1 ovarian and (E) T47D-KBluc breast cancer cells. Cells were treated with 100 nM NC or IMP1 siRNA SmartPool for 72 hours prior to evaluating cell number (n=6). “•” denotes number of cells plated at start of experiment. BTYNB blocks proliferation of (F) IGROV-1, (G) ES-2, and (H) SK-MEL2 cancer cells with no effects on (I) IMP1-negative BG-1 ovarian and T47D-KBluc breast cancer cells. Cells were treated with a DMSO-control or increasing concentrations of BTYNB for 72 hours prior to evaluating cell number (n=6). (J) The biologically inactive compound, 5226752, has no effects on proliferation of IMP1-positive and IMP1-negative cancer cell lines. Cells were treated for 72 hours with increasing concentrations of compound 5226752 (n=6). (K) Overexpression of IMP1 is sufficient to block BTYNB-mediated inhibition of cell proliferation (n=6). IMP1-positive HEK 293T cells were transiently transfected with 100 ng of a control-plasmid (Control) or CMV-IMP1 expression plasmid (IMP1 overexpression). Western blot shows increased IMP1 protein in the cells transfected with CMV-IMP1. Twenty four hours after transfection, cells were treated with the control (DMSO) or 10 μM BTYNB for 48 hours before evaluating cell number. Proliferation rates were normalized to cells treated with DMSO-control. Data are reported as mean±SEM and P values represent comparison to NC controls (A-E). *** P<.001. ns, not significant. Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions

Figure 6 BTYNB inhibits anchorage-independent growth of IMP1-positive cancer cells. (A) SK-MEL2 cells treated for 21 days with medium containing DMSO-vehicle control or 10 μM BTYNB. (B) ES-2 cells treated for 14 days with medium containing DMSO-vehicle control or 10 μM BTYNB. Photographs are at 10× magnification and are representative of the entire plate and of triplicate experiments. The bar graphs represent the average of the total number of colonies counted after treating with DMSO-control or 10 μM BTYNB. ***P<.001. Translational Oncology 2017 10, 818-827DOI: (10.1016/j.tranon.2017.07.008) Copyright © 2017 The Authors Terms and Conditions