Re-Expression of the Retinoblastoma-Binding Protein 2-Homolog 1 Reveals Tumor- Suppressive Functions in Highly Metastatic Melanoma Cells Alexander Roesch,

Slides:

Advertisements

Similar presentations

Juewon Kim, Hyunjung Choi, Eun-Gyung Cho, Tae R. Lee

Advertisements

Volume 26, Issue 3, Pages (March 2007)

Membrane-Tethered Intracellular Domain of Amphiregulin Promotes Keratinocyte Proliferation Stefan W. Stoll, Philip E. Stuart, Sylviane Lambert, Alberto.

Volume 81, Issue 1, Pages (January 2012)

Pericentrosomal Localization of the TIG3 Tumor Suppressor Requires an N-Terminal Hydrophilic Region Motif Tiffany M. Scharadin, Gautam Adhikary, Kristin.

Mechanisms of p53 Restriction in Merkel Cell Carcinoma Cells Are Independent of the Merkel Cell Polyoma Virus T Antigens Roland Houben, Christina Dreher,

MAGE-C2 Promotes Growth and Tumorigenicity of Melanoma Cells, Phosphorylation of KAP1, and DNA Damage Repair Neehar Bhatia, Tony Z. Xiao, Kimberly A.

Dominant Effects of Δ40p53 on p53 Function and Melanoma Cell Fate

Modification of Alternative Splicing of Mcl-1 Pre-mRNA Using Antisense Morpholino Oligonucleotides Induces Apoptosis in Basal Cell Carcinoma Cells Jeng-Jer.

Shitao Li, Lingyan Wang, Michael A. Berman, Ye Zhang, Martin E. Dorf

Mechanisms of p53 Restriction in Merkel Cell Carcinoma Cells Are Independent of the Merkel Cell Polyoma Virus T Antigens Roland Houben, Christina Dreher,

Naoko Kanda, Shinichi Watanabe Journal of Investigative Dermatology

SOX10 Promotes Melanoma Cell Invasion by Regulating Melanoma Inhibitory Activity Saskia A. Graf, Christian Busch, Anja-Katrin Bosserhoff, Robert Besch,

The DNA-Mismatch Repair Enzyme hMSH2 Modulates UV-B-Induced Cell Cycle Arrest and Apoptosis in Melanoma Cells Markus Seifert, Stefan J. Scherer, Wilfried.

Complexity of VEGF Responses in Skin Carcinogenesis Revealed through Ex Vivo Assays Based on a VEGF-A Null Mouse Keratinocyte Cell Line Isabel Mirones,

UCHL1 Regulates Melanogenesis through Controlling MITF Stability in Human Melanocytes Eun Young Seo, Seon-Pil Jin, Kyung-Cheol Sohn, Chi-Hyun Park, Dong.

Low Concentrations of Curcumin Induce Growth Arrest and Apoptosis in Skin Keratinocytes Only in Combination with UVA or Visible Light Jadranka Dujic,

Stefan W. Stoll, Jessica L. Johnson, Yong Li, Laure Rittié, James T

Tumor-Derived Fibronectin Is Involved in Melanoma Cell Invasion and Regulated by V600E B-Raf Signaling Pathway Cédric Gaggioli, Guillaume Robert, Corine.

Multiple PKCδ Tyrosine Residues Are Required for PKCδ-Dependent Activation of Involucrin Expression—a Key Role of PKCδ-Y311 Ling Zhu, Chaya Brodie, Sivaprakasam.

Ganglioside GM3 Promotes Carcinoma Cell Proliferation via Urokinase Plasminogen Activator-Induced Extracellular Signal-Regulated Kinase-Independent p70S6.

High-Efficiency Stable Gene Transduction in Primary Human Melanocytes Using a Lentiviral Expression System Shariff Dunlap, Xiaobing Yu, Linzhao Cheng,

Decreased Growth Inhibitory Responses of Squamous Carcinoma Cells to Interferon-γ Involve Failure to Recruit cki Proteins into cdk2 Complexes Beth L.

Volume 7, Issue 6, Pages (June 2005)

Claudia D. Andl, John R. Stanley Journal of Investigative Dermatology

Volume 21, Issue 1, Pages (January 2006)

Merkel Cell Polyomavirus–Positive Merkel Cell Carcinoma Requires Viral Small T- Antigen for Cell Proliferation Masahiro Shuda, Yuan Chang, Patrick S.

MiTF Regulates Cellular Response to Reactive Oxygen Species through Transcriptional Regulation of APE-1/Ref-1 Feng Liu, Yan Fu, Frank L. Meyskens Journal.

Enhanced Death Ligand-Induced Apoptosis in Cutaneous SCC Cells by Treatment with Diclofenac/Hyaluronic Acid Correlates with Downregulation of c-FLIP

Inhibition of Human Melanoma Cell Growth by the Dietary Flavonoid Fisetin Is Associated with Disruption of Wnt/β-Catenin Signaling and Decreased Mitf.

Selective Induction of Apoptosis in Melanoma Cells by Tyrosinase Promoter-Controlled CD95 Ligand Overexpression Lothar F. Fecker, Christoph C. Geilen,

Induction of Terminal Differentiation in Melanoma Cells on Downregulation of β-Amyloid Precursor Protein Michelle G. Botelho, Xiaolei Wang, Donna J.

The Effects of Dickkopf 1 on Gene Expression and Wnt Signaling by Melanocytes: Mechanisms Underlying Its Suppression of Melanocyte Function and Proliferation

Epigenetic Control of MAGE Gene Expression by the KIT Tyrosine Kinase

Lubrol-RAFTs in Melanoma Cells: A Molecular Platform for Tumor-Promoting Ephrin-B2– Integrin-β1 Interaction Stefanie Meyer, Evelyn Orsó, Gerd Schmitz,

Noah C. Jenkins, Jae Jung, Tong Liu, Megan Wilde, Sheri L

Volume 8, Issue 1, Pages (January 1998)

Michael P. O'Connell, Ashani T. Weeraratna

Volume 13, Issue 6, Pages (June 2013)

Oncogenic B-RAFV600E Signaling Induces the T-Box3 Transcriptional Repressor to Repress E-Cadherin and Enhance Melanoma Cell Invasion Suzanah C. Boyd,

Pituitary Tumor-Transforming Gene 1 Enhances Proliferation and Suppresses Early Differentiation of Keratinocytes Yosuke Ishitsuka, Yasuhiro Kawachi,

ERK1/2 Is Highly Phosphorylated in Melanoma Metastases and Protects Melanoma Cells from Cisplatin-Mediated Apoptosis Alireza Mirmohammadsadegh, Rodrigo.

Volume 56, Issue 5, Pages (December 2014)

Volume 13, Issue 4, Pages (April 2013)

Interleukin-6-Resistant Melanoma Cells Exhibit Reduced Activation of STAT3 and Lack of Inhibition of Cyclin E-Associated Kinase Activity Markus Böhm,

Protein Kinase C-βII Represses Hepatocyte Growth Factor-Induced Invasion by Preventing the Association of Adapter Protein Gab1 and Phosphatidylinositol.

Romain Debret, Richard R

Arsenic Induces Tumor Necrosis Factor α Release and Tumor Necrosis Factor Receptor 1 Signaling in T Helper Cell Apoptosis Hsin-Su Yu, Gwo-Shing Chen

Wnt Signaling through the β-Catenin Pathway Is Sufficient to Maintain, but Not Restore, Anagen-Phase Characteristics of Dermal Papilla Cells Hidenao.

The p73 Gene Is an Anti-Tumoral Target of the RARβ/γ-Selective Retinoid Tazarotene Marina Papoutsaki, Mauro Lanza, Barbara Marinari, Steven Nisticò, Francesca.

Insulin-Like Growth Factor-Binding Protein 7 Regulates Keratinocyte Proliferation, Differentiation and Apoptosis Janna Nousbeck, Ofer Sarig, Nili Avidan,

Hydroxychloroquine Modulates Metabolic Activity and Proliferation and Induces Autophagic Cell Death of Human Dermal Fibroblasts Bettina Ramser, Agatha.

Activation of Dual Apoptotic Pathways in Human Melanocytes and Protection by Survivin Tong Liu, Diana Biddle, Adrianne N. Hanks, Brook Brouha, Hui Yan,

Ruth Halaban, Elaine Cheng Journal of Investigative Dermatology

Discrimination of Melanocytic Tumors by cDNA Array Hybridization of Tissues Prepared by Laser Pressure Catapulting Bernd Becker, Alexander Roesch, Christian.

Meghan E. Hermance, Saravanan Thangamani

Interferon-γ-Mediated Growth Regulation of Melanoma Cells: Involvement of STAT1- Dependent and STAT1-Independent Signals Anja Bosserhoff Journal of Investigative.

Transcriptional Repression of miR-34 Family Contributes to p63-Mediated Cell Cycle Progression in Epidermal Cells Dario Antonini, Monia T. Russo, Laura.

MELK Promotes Melanoma Growth by Stimulating the NF-κB Pathway

Mechanical Stretching In Vitro Regulates Signal Transduction Pathways and Cellular Proliferation in Human Epidermal Keratinocytes Shoichiro Yano, Mayumi.

ΔNp63α Promotes Apoptosis of Human Epidermal Keratinocytes

Functional Modulation of IGF-Binding Protein-3 Expression in Melanoma

Merkel Cell Polyomavirus–Positive Merkel Cell Carcinoma Cells Do Not Require Expression of the Viral Small T Antigen Sabrina Angermeyer, Sonja Hesbacher,

John W. Haycock, Mark Wagner, Sheila Mac Neil

The Tumor Suppressor PP2A Aβ Regulates the RalA GTPase

Mahesh Ramamoorthy, Susan Smith Cancer Cell

Sphingosine 1-Phosphate Restrains Insulin-Mediated Keratinocyte Proliferation via Inhibition of Akt through the S1P2 Receptor Subtype Melanie Schüppel,

Epidermal Growth Factor Receptor Pathway Mitigates UVA-Induced G2/M Arrest in Keratinocyte Cells Christine Jean, Hélène Hernandez-Pigeon, Amandine Blanc,

Ultraviolet-B-Induced G1 Arrest is Mediated by Downregulation of Cyclin-Dependent Kinase 4 in Transformed Keratinocytes Lacking Functional p53 Arianna.

Volume 11, Issue 7, Pages (July 2018)

Presentation transcript:

Re-Expression of the Retinoblastoma-Binding Protein 2-Homolog 1 Reveals Tumor- Suppressive Functions in Highly Metastatic Melanoma Cells Alexander Roesch, Bernd Becker, Wulf Schneider-Brachert, Ilja Hagen, Michael Landthaler, Thomas Vogt Journal of Investigative Dermatology Volume 126, Issue 8, Pages 1850-1859 (August 2006) DOI: 10.1038/sj.jid.5700324 Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Lentiviral construct. The C-term of RBP2-H1 containing the NTE1A of RBP2-H1 (aa 1391–1490) and a LMO2-binding domain (aa 1536–1574) was subcloned into the lentiviral expression vector pQCXIP finally coding for a FLAG-tagged cRBP2-H1 fusion protein. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Lentiviral re-expression of cRBP2-H1. In contrast to uninfected A375-SM melanoma cells and cells infected with a lentiviral construct coding for GFP, exclusively in cRBP2-H1-transduced cells the specific anti-FLAG antibody could detect the presumed band of 22.58kDa. 20ng of FLAG-BAP™ fusion protein served as antibody control. PDI was used as loading control. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Co-immunoprecipitation of pRb with cRBP2-H1. In contrast to uninfected A375-SM melanoma cells and cells infected with the GFP control, exclusively in cRBP2-H1-transduced cells a co-immunoprecipitation of pRb with expressed cRBP2-H1 was seen. Co-immunoprecipitation succeeded in both directions, that is precipitation with anti-FLAG and detection with anti-pRb antibody (upper row) or precipitation with anti-pRb and detection with anti-FLAG antibody (lower row). Unspecific signals larger in size than the fusion peptide are marked with an asterisk. They fail to show any co-immunoprecipitation. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Phospho-pRb immunoblot. (a) In lentivirus-transduced A375-SM cells selectively re-expressing cRBP2-H1, a stabilizing potential on hypophosphorylated Ser795 was detected 3 and 9hours after release from colchicine synchronization. Phosphorylation was compared to untreated A375-SM cells and cells infected with a control virus coding for a non-cell-cycle-related protein (GFP). At Ser780 and Ser807/811, no change in pRb phosphorylation was observed. Total pRb and PDI expression remained constant. Double bands result from differently phosphorylated pRb molecules. (b) A375-SM cells transfected with the N-term of RBP2-H1 lacking the NTE1A fails to stabilize hypophosphorylated pRb at each of the analyzed phospho-sites. No differences could be seen compared to mock-transfected A375-SM cells. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Control immunoblot. CyclinD1 and cdk4 protein expression levels remained unchanged between cRBP2-H1-transduced cells, untreated A375-SM cells, and cells infected with the control virus. PDI expression also remained constant. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Cell cycle analysis. (a) Cell cycle profiling with propidium iodide (FACS).In contrast to native A375-SM cells (dark blue) and control virus-infected cells (black), cRBP2-H1-transduced A375-SM cells (light blue) show a highly significant shift from S- and G2/M phase towards G1 phase (G1 arrest). The cell cycle profiles of non-infected A375-SM and control virus cells were almost congruent. Depicted is the result of one representative experiment out of six independently performed experiments. (b) BrdU incorporation into replicated DNA (Cell Proliferation ELISA™). As a consequence of the observed G1 arrest, cRBP2-H1-transduced cells exhibit significantly reduced BrdU incorporation during the S phase compared to both controls. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 AlamarBlue™ fluorescence assay. Compared to native A375-SM cells and control virus-infected cells, cRBP2-H1-re-expressing A375-SM cells show a highly significant reduction in cellular proliferation, probably due to G1 arrest. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 cRBP2-H1-induced gene regulation. Combining Ingenuity™ network analysis with conventional literature research, two networks including melanoma-relevant genes, as for example MITF, dopachrome tautomerase, tyrosinase-related protein 1, TCF4, or HGF (hepatocyte growth factor) could be linked using SPI1 (spleen focus forming virus proviral integration oncogene spi1, PU.1, Tcfpu.1) as a branching point. Moreover, screening the list of filtered genes manually, also typically E2F-regulated genes, as for example follistatin (follistatin), bone morphogenetic protein 2 (BHP-2), and transforming growth factor alpha were found to be regulated. White symbols: no regulation found, green: downregulation, red: upregulation. The depicted value represents the regulation cRBP2-H1/NTE1A versus GFP control. A: activation, B: binding, E: expression. Journal of Investigative Dermatology 2006 126, 1850-1859DOI: (10.1038/sj.jid.5700324) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions