Sarah A. Best, Amy N. Nwaobasi, Chrysalyne D. Schmults, Matthew R

Slides:

Advertisements

Similar presentations

Cell Physiol Biochem 2013;32: DOI: /

Advertisements

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector Joanna Jacków, Matthias.

An Acrodermatitis Enteropathica-Associated Zn Transporter, ZIP4, Regulates Human Epidermal Homeostasis Bum-Ho Bin, Jinhyuk Bhin, Nan-Hyung Kim, Su-Hyon.

Cell Surface CD74–MIF Interactions Drive Melanoma Survival in Response to Interferon-γ Keiji Tanese, Yuuri Hashimoto, Zuzana Berkova, Yuling Wang, Felipe.

IFN-γ Primes Keratinocytes for HSV-1–Induced Inflammasome Activation

Maik Dahlhoff, Sukalp Muzumdar, Matthias Schäfer, Marlon R. Schneider

PFKFB3, a Direct Target of p63, Is Required for Proliferation and Inhibits Differentiation in Epidermal Keratinocytes Robert B. Hamanaka, Gökhan M. Mutlu

Phenformin Inhibits Myeloid-Derived Suppressor Cells and Enhances the Anti-Tumor Activity of PD-1 Blockade in Melanoma Sun Hye Kim, Man Li, Sebastian.

Jasmine George, Minakshi Nihal, Chandra K

MicroRNA-31 Promotes Skin Wound Healing by Enhancing Keratinocyte Proliferation and Migration Dongqing Li, X.I. Li, Aoxue Wang, Florian Meisgen, Andor.

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

KIND1 Loss Sensitizes Keratinocytes to UV-Induced Inflammatory Response and DNA Damage Xiaoling Zhang, Suju Luo, Joseph Wu, Long Zhang, Wen-hui Wang,

Research Techniques Made Simple: Identification and Characterization of Long Noncoding RNA in Dermatological Research Dario Antonini, Maria Rosaria Mollo,

NADPH Oxidase-1 Plays a Key Role in Keratinocyte Responses to UV Radiation and UVB-Induced Skin Carcinogenesis Houssam Raad, Martin Serrano-Sanchez,

Sarah A. Best, Amy N. Nwaobasi, Chrysalyne D. Schmults, Matthew R

Development of Cell-Penetrating Asymmetric Interfering RNA Targeting Connective Tissue Growth Factor Jihye Hwang, Chanil Chang, Ji Hyun Kim, Chang Taek.

Dual Role of Apoptosis-Associated Speck-Like Protein Containing a CARD (ASC) in Tumorigenesis of Human Melanoma Weimin Liu, Yuchun Luo, Jeffrey H. Dunn,

Ann E. Collier, Ronald C. Wek, Dan F. Spandau

Regulation and Function of the Caspase-1 in an Inflammatory Microenvironment Dai-Jen Lee, Fei Du, Shih-Wei Chen, Manando Nakasaki, Isha Rana, Vincent.

CtBP1 Overexpression in Keratinocytes Perturbs Skin Homeostasis

Trim32 Deficiency Enhances Th2 Immunity and Predisposes to Features of Atopic Dermatitis Yuangang Liu, Zhiping Wang, Rachel De La Torre, Ashley Barling,

Molecular Therapy - Nucleic Acids

Maik Dahlhoff, Sukalp Muzumdar, Matthias Schäfer, Marlon R. Schneider

Yes-Associated Protein Contributes to the Development of Human Cutaneous Squamous Cell Carcinoma via Activation of RAS Jinjing Jia, Changji Li, Suju.

Overexpression of PRAS40T246A in the Proliferative Compartment Suppresses mTORC1 Signaling, Keratinocyte Migration, and Skin Tumor Development Okkyung.

Volume 6, Issue 1, Pages (January 2014)

HPV8 Field Cancerization in a Transgenic Mouse Model Is due to Lrig1+ Keratinocyte Stem Cell Expansion Simone Lanfredini, Carlotta Olivero, Cinzia Borgogna,

Nicastrin/miR-30a-3p/RAB31 Axis Regulates Keratinocyte Differentiation by Impairing EGFR Signaling in Familial Acne Inversa Yanyan He, Haoxiang Xu, Chengrang.

Smad4 Loss in Mouse Keratinocytes Leads to Increased Susceptibility to UV Carcinogenesis with Reduced Ercc1-Mediated DNA Repair Doyel Mitra, Pamela Fernandez,

Transdifferentiation of Melanoma Cells by the Reprogramming Factors Attenuates Malignant Nature In Vitro and In Vivo Mikiro Takaishi, Shigetoshi Sano

Maria Rosaria Mollo, Dario Antonini, Luisa Cirillo, Caterina Missero

Transcription Factor MafB Coordinates Epidermal Keratinocyte Differentiation Masashi Miyai, Michito Hamada, Takashi Moriguchi, Junichiro Hiruma, Akiyo.

Spleen Tyrosine Kinase Mediates EGFR Signaling to Regulate Keratinocyte Terminal Differentiation Nan-Lin Wu, Duen-Yi Huang, Li-Fang Wang, Reiji Kannagi,

Christina A. Young, Richard L

Epidermal Desmoglein 1 Expression Is Reduced in Kidney Transplant Recipients Compared with Immunocompetent Patients Jodi L. Johnson, Paul Hoover, Borko.

P63 Transcription Factor Regulates Nuclear Shape and Expression of Nuclear Envelope-Associated Genes in Epidermal Keratinocytes Valentina Rapisarda,

Skin-Specific Deletion of Mis18α Impedes Proliferation and Stratification of Epidermal Keratinocytes Koog Chan Park, Minkyoung Lee, Yoon Jeon, Raok Jeon,

IQGAP1 and IQGAP3 Serve Individually Essential Roles in Normal Epidermal Homeostasis and Tumor Progression Christine L. Monteleon, Andrew McNeal, Elizabeth.

c-Jun Promotes whereas JunB Inhibits Epidermal Neoplasia

SOX4 Promotes Proliferative Signals by Regulating Glycolysis through AKT Activation in Melanoma Cells Wei Dai, Xinyuan Xu, Shuli Li, Jingjing Ma, Qiong.

CCN1, a Pro-Inflammatory Factor, Aggravates Psoriasis Skin Lesions by Promoting Keratinocyte Activation Yue Sun, Jie Zhang, Zhou Zhou, Pinru Wu, Rongfen.

Inhibition of KLF4 by Statins Reverses Adriamycin-Induced Metastasis and Cancer Stemness in Osteosarcoma Cells Yangling Li, Miao Xian, Bo Yang, Meidan.

Wei Xu, Shengxian Jia, Ping Xie, Aimei Zhong, Robert D

Brian Poligone, Elaine S. Gilmore, Carolina V

Regulation of Fibulin-2 Gene Expression by Integrin α3β1 Contributes to the Invasive Phenotype of Transformed Keratinocytes Dara S. Missan, Sridar V.

SIRT1 Activation Ameliorates Aldara-Induced Psoriasiform Phenotype and Histology in Mice Sijing Xie, Zhonglan Su, Bin Zhang, Jiuyu Ge, Shiyu Song, Guibo.

Human Mitochondrial NAD(P)+–Dependent Malic Enzyme Participates in Cutaneous Melanoma Progression and Invasion Yung-Lung Chang, Hong-Wei Gao, Chien-Ping.

GLI2 Is a Regulator of β-Catenin and Is Associated with Loss of E-Cadherin, Cell Invasiveness, and Long-Term Epidermal Regeneration Eleni Pantazi, Emilios.

Lack of Collagen VI Promotes Wound-Induced Hair Growth

Overexpression of CD109 in the Epidermis Differentially Regulates ALK1 Versus ALK5 Signaling and Modulates Extracellular Matrix Synthesis in the Skin

Collagen VII Half-Life at the Dermal-Epidermal Junction Zone: Implications for Mechanisms and Therapy of Genodermatoses Tobias Kühl, Markus Mezger, Ingrid.

14-3-3σ Regulates Keratinocyte Proliferation and Differentiation by Modulating Yap1 Cellular Localization Sumitha A.T. Sambandam, Ramesh B. Kasetti,

Volume 6, Issue 1, Pages (January 2014)

Dual Role of the Anaphase Promoting Complex/Cyclosome in Regulating Stemness and Differentiation in Human Primary Keratinocytes Ling Shih Quek, Nicolas.

Nrf2 Promotes Keratinocyte Proliferation in Psoriasis through Up-Regulation of Keratin 6, Keratin 16, and Keratin 17 Luting Yang, Xueli Fan, Tingting.

Tej Pratap Singh, Gerlinde Mayer, Peter Wolf

dsRNA Sensing Induces Loss of Cell Identity

TSLP Is a Potential Initiator of Collagen Synthesis and an Activator of CXCR4/SDF-1 Axis in Keloid Pathogenesis Jung U Shin, Seo Hyeong Kim, Hyeran Kim,

Jasmine George, Minakshi Nihal, Chandra K

Integrin-β4–TNS4–Focal Adhesion Kinase Signaling Mediates Keratinocyte Proliferation in Human Skin Eun Young Seo, Seon-Pil Jin, Yeon Kyung Kim, Hanon.

Macrophage Inhibitory Cytokine-1 Is Overexpressed in Malignant Melanoma and Is Associated with Tumorigenicity Glen M. Boyle, Julie Pedley, Adam C. Martyn,

UVA Induces Lesions Resembling Seborrheic Keratoses in Mice with Keratinocyte- Specific PTEN Downregulation Mei Ming, Christopher R. Shea, Li Feng, Keyoumars.

Anna Flammiger, Robert Besch, Anthony L. Cook, Tanja Maier, Richard A

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector Joanna Jacków, Matthias.

MELK Promotes Melanoma Growth by Stimulating the NF-κB Pathway

Javed Mohammed, Andrew Ryscavage, Rolando Perez-Lorenzo, Andrew J

IL-17A Upregulates Keratin 17 Expression in Keratinocytes through STAT1- and STAT3- Dependent Mechanisms Xiaowei Shi, Liang Jin, Erle Dang, Ting Chang,

Volume 5, Issue 6, Pages (December 2013)

Shipra Das, Olga Anczuków, Martin Akerman, Adrian R. Krainer

Anticancer Activity of the Cholesterol Exporter ABCA1 Gene

Presentation transcript:

CCAR2 Is Required for Proliferation and Tumor Maintenance in Human Squamous Cell Carcinoma Sarah A. Best, Amy N. Nwaobasi, Chrysalyne D. Schmults, Matthew R. Ramsey Journal of Investigative Dermatology Volume 137, Issue 2, Pages 506-512 (February 2017) DOI: 10.1016/j.jid.2016.09.027 Copyright © 2016 The Authors Terms and Conditions

Figure 1 CCAR2 and SIRT1 proteins are stabilized in squamous cell carcinoma. (a) Histologic (H&E) and immunofluorescence analysis of CCAR2 (green) and keratin 14 (K14, red) protein expression in normal human skin and SCC frozen sections. DAPI (blue) stains the nucleus. Scale bar = 50 μm. (b) Western blot analysis of CCAR2 and SIRT1 protein in primary keratinocytes (OKF6, P1-Ep, and N) and SCC (SCC-13, SCC-25, JHU-029, and HO1N1) cell lines. GAPDH provides the loading control. (c) Quantitative real-time PCR of CCAR2 (left) and SIRT1 (right) mRNA expression in keratinocytes and SCC cell lines. P > 0.05, not significant. (d) Western blot of CCAR2 and SIRT1 protein expression in primary keratinocytes (N, P1-Ep) and SCC (HO1N1 and JHU-029) cell lines treated with cyclohexamide (100 μg/ml) for 0, 2, 4, 6, or 8 hours. GAPDH provides the loading control. CHX, cyclohexamide; H&E, hematoxylin and eosin; K14, keratin 14; N.S., not significant; SCC, squamous cell carcinoma. Journal of Investigative Dermatology 2017 137, 506-512DOI: (10.1016/j.jid.2016.09.027) Copyright © 2016 The Authors Terms and Conditions

Figure 2 CCAR2 is essential for proliferation and tumor maintenance. (a) Flow cytometry analysis of the cell cycle (G1, S, or G2/M) in control (Empty) or CCAR2-knockdown (shCCAR2) SCC cell lines. N = 3 replicates. Error bars indicate ± standard deviation. Representative from two independent repeats. (b) Colony assay in control or CCAR2 knockdown SCC cell lines. Colony number is relative to the control of each cell line. Error bars indicate ± standard deviation. N = 4 replicates per cell line. (c) Growth of xenograft tumors derived from JHU-029 cells with doxycycline-inducible shRNA, injected subcutaneously in nude mice. Doxycycline was added to drinking water on day 4. Tumor volume is normalized to average day 6 tumor volume for each group. Error bars ± standard error of the mean. (d) Western blot analysis of CCAR2 protein in JHU-029 xenograft tumors with indicated treatment as in c. (e) Analysis of proliferative index of tumors from c at day 18 as assessed by Ki67 staining. *P < 0.05, **P < 0.01, ***P < 0.001 for all experiments. DOX, doxycycline; SCC, squamous cell carcinoma. Journal of Investigative Dermatology 2017 137, 506-512DOI: (10.1016/j.jid.2016.09.027) Copyright © 2016 The Authors Terms and Conditions

Figure 3 CCAR2 regulates genes involved in cell division. (a) Heat map depicting gene expression changes from microarray in CCAR2 knockdown (shCCAR2) compared with control cells (Empty) in SCC cell lines (SCC-13, HO1N1, SCC-15). (b) Quantitative real-time PCR validation of genes identified by the microarray in SCC-13 (top panel) and HO1N1 cells (bottom panel) with either control (Empty) or CCAR2 knockdown (shCCAR2). ∗∗P < 0.01, ∗∗∗P < 0.001. Journal of Investigative Dermatology 2017 137, 506-512DOI: (10.1016/j.jid.2016.09.027) Copyright © 2016 The Authors Terms and Conditions

Figure 4 CCAR2 regulates stability of acetylated transcription factors in SCC. (a) Western blot analysis of CCAR2, RFX1, and CREB protein in SCC cell lines with CCAR2 knockdown (shCCAR2) or control (Empty). GAPDH provides the loading control. (b) Western blot analysis of RFX1 and CREB protein in keratinocytes and SCC cells. GAPDH provides the loading control. (c) Immunoprecipitation of acetylated RFX1 protein from whole-cell extracts of SCC-13 cells. IgG serves as a loading control for IP. (d) Co-immunoprecipitation of RFX1 and SIRT1 proteins from whole-cell extracts of SCC-13. IgG serves as a loading control for IP. (e) Co-immunoprecipitation of RFX1 and CCAR2 proteins from whole-cell extracts of SCC-13. IgG serves as a loading control for IP. IP, immunoprecipication; SCC, squamous cell carcinoma. Journal of Investigative Dermatology 2017 137, 506-512DOI: (10.1016/j.jid.2016.09.027) Copyright © 2016 The Authors Terms and Conditions

Figure 5 RFX1 and CREB are required for proliferation in SCC. (a) Flow cytometry analysis of the cell cycle (G1, S or G2/M) in control (Empty), RFX1 knockdown (shRFX1), in JHU-029 cells (n = 3). (b) Flow cytometry analysis of the cell cycle in control (Empty) or CREB knockdown (shCREB) in JHU-029 cells (n = 3). Error bars indicate ± standard deviation. (c) Colony assay in control (Empty), RFX1 knockdown (shRFX1) in JHU-029 cells. Colony number is relative to the control of each cell line. Error bars indicate ± standard deviation. N = 4 replicates per cell line. Representative from two independent repeats. (d) Colony assay in control (Empty) CREB knockdown (shCREB) in JHU-029 cells. SCC, squamous cell carcinoma. *P < 0.05, **P < 0.01, ***P < 0.001. Journal of Investigative Dermatology 2017 137, 506-512DOI: (10.1016/j.jid.2016.09.027) Copyright © 2016 The Authors Terms and Conditions