Abstract
The matricellular protein CCN5/WISP-2 represents a promising target in triple-negative breast cancer (TNBC) because treatment or induced activation of CCN5 in TNBC cells promotes cell growth arrest at the G0/G1 phase, reduces cell proliferation and delays tumor growth in the xenograft model. Our studies found that the p27Kip1 tumor suppressor protein is upregulated and relocalized to the nucleus from cytoplasm by CCN5 in these cells and that these two events (upregulation and relocalization of p27Kip1) are critical for CCN5-induced growth inhibition of TNBC cells. In the absence of CCN5, p27Kip1 resides mostly in the cytoplasm, which is associated with the aggressive nature of cancer cells. Mechanistically, CCN5 inhibits Skp2 expression, which seems to stabilize the p27Kip1 protein in these cells. On the other hand, CCN5 also recruits FOXO3a to mediate the transcriptional regulation of p27Kip1. The recruitment of FOXO3a is achieved by the induction of its expression and activity through shifting from cytoplasm to the nucleus. Our data indicate that CCN5 blocks PI3K/AKT signaling to dephosphorylate at S318, S253 and Thr32 in FOXO3a for nuclear relocalization and activation of FOXO3a. Moreover, inhibition of α6β1 receptors diminishes CCN5 action on p27Kip1 in TNBC cells. Collectively, these data suggest that CCN5 effectively inhibits TNBC growth through the accumulation and trafficking of p27Kip1 via Skp2 and FOXO3a regulation, and thus, activation of CCN5 may have the therapeutic potential to kill TNBC.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Oliveira LR, Jeffrey SS, Ribeiro-Silva A . Stem cells in human breast cancer. Histol Histopathol 2010; 25: 371–385.
Simpson PT, Gale T, Reis-Filho JS, Jones C, Parry S, Sloane JP et al. Columnar cell lesions of the breast: the missing link in breast cancer progression? A morphological and molecular analysis. Am J Surg Pathol 2005; 29: 734–746.
Banerjee SK, Banerjee S . CCN5/WISP-2: A micromanager of breast cancer progression. J Cell Commun Signal 2012; 6: 63–71.
Fritah A, Saucier C, De WO, Bracke M, Bieche I, Lidereau R et al. Role of WISP-2/CCN5 in the maintenance of a differentiated and noninvasive phenotype in human breast cancer cells. Mol Cell Biol 2008; 28: 1114–1123.
Dhar G, Mehta S, Banerjee S, Gardner A, McCarty BM, Mathur SC et al. Loss of WISP-2/CCN5 signaling in human pancreatic cancer: a potential mechanism for epithelial-mesenchymal-transition. Cancer Lett 2007; 254: 63–70.
Sabbah M, Prunier C, Ferrand N, Megalophonos V, Lambein K, De Wever O et al. CCN5, a novel transcriptional repressor of the transforming growth factor beta signaling pathway. Mol Cell Biol 2011; 31: 1459–1469.
Haque I, Banerjee S, Mehta S, De A, Majumder M, Mayo MS et al. Cysteine-rich 61-connective tissue growth factor-nephroblastoma-overexpressed 5 (CCN5)/Wnt-1-induced signaling protein-2 (WISP-2) regulates microRNA-10b via hypoxia-inducible factor-1alpha-TWIST signaling networks in human breast cancer cells. J Biol Chem 2011; 286: 43475–43485.
Banerjee S, Dhar G, Haque I, Kambhampati S, Mehta S, Sengupta K et al. CCN5/WISP-2 expression in breast adenocarcinoma is associated with less frequent progression of the disease and suppresses the invasive phenotypes of tumor cells. Cancer Res 2008; 68: 7606–7612.
Dhar G, Banerjee S, Dhar K, Tawfik O, Mayo MS, Vanveldhuizen PJ et al. Gain of oncogenic function of p53 mutants induces invasive phenotypes in human breast cancer cells by silencing CCN5/WISP-2. Cancer Res 2008; 68: 4580–4587.
Besson A, Dowdy SF, Roberts JM . CDK inhibitors: cell cycle regulators and beyond. Dev Cell 2008; 14: 159–169.
Alkarain A, Jordan R, Slingerland J . p27 deregulation in breast cancer: prognostic significance and implications for therapy. J Mammary Gland Biol Neoplasia 2004; 9: 67–80.
Besson A, Assoian RK, Roberts JM . Regulation of the cytoskeleton: an oncogenic function for CDK inhibitors? Nat Rev Cancer 2004; 4: 948–955.
Sherr CJ, Roberts JM . CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 1999; 13: 1501–1512.
Coqueret O . New roles for p21 and p27 cell-cycle inhibitors: a function for each cell compartment? Trends Cell Biol 2003; 13: 65–70.
Mineta H, Miura K, Suzuki I, Takebayashi S, Amano H, Araki K et al. Low p27 expression correlates with poor prognosis for patients with oral tongue squamous cell carcinoma. Cancer 1999; 85: 1011–1017.
Chu IM, Hengst L, Slingerland JM . The Cdk inhibitor p27 in human cancer: prognostic potential and relevance to anticancer therapy. Nat Rev Cancer 2008; 8: 253–267.
Toyoshima H, Hunter T . p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21. Cell 1994; 78: 67–74.
Wander SA, Zhao D, Slingerland JM . p27: a barometer of signaling deregulation and potential predictor of response to targeted therapies. Clin Cancer Res 2011; 17: 12–18.
Nourse J, Firpo E, Flanagan WM, Coats S, Polyak K, Lee MH et al. Interleukin-2-mediated elimination of the p27Kip1 cyclin-dependent kinase inhibitor prevented by rapamycin. Nature 1994; 372: 570–573.
Polyak K, Kato JY, Solomon MJ, Sherr CJ, Massague J, Roberts JM et al. p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev 1994; 8: 9–22.
Bloom J, Pagano M . Deregulated degradation of the cdk inhibitor p27 and malignant transformation. Semin Cancer Biol 2003; 13: 41–47.
Slingerland J, Pagano M . Regulation of the cdk inhibitor p27 and its deregulation in cancer. J Cell Physiol 2000; 183: 10–17.
Baldassarre G, Belletti B, Bruni P, Boccia A, Trapasso F, Pentimalli F et al. Overexpressed cyclin D3 contributes to retaining the growth inhibitor p27 in the cytoplasm of thyroid tumor cells. J Clin Invest 1999; 104: 865–874.
Sgambato A, Cittadini A, Faraglia B, Weinstein IB . Multiple functions of p27(Kip1) and its alterations in tumor cells: a review. J Cell Physiol 2000; 183: 18–27.
Asada M, Yamada T, Ichijo H, Delia D, Miyazono K, Fukumuro K et al. Apoptosis inhibitory activity of cytoplasmic p21(Cip1/WAF1) in monocytic differentiation. EMBO J 1999; 18: 1223–1234.
Blagosklonny MV . Are p27 and p21 cytoplasmic oncoproteins? Cell Cycle 2002; 1: 391–393.
Motti ML, Califano D, Troncone G, De Marco C, Migliaccio I, Palmieri E et al. Complex regulation of the cyclin-dependent kinase inhibitor p27kip1 in thyroid cancer cells by the PI3K/AKT pathway: regulation of p27kip1 expression and localization. Am J Pathol 2005; 166: 737–749.
Tan P, Cady B, Wanner M, Worland P, Cukor B, Magi-Galluzzi C et al. The cell cycle inhibitor p27 is an independent prognostic marker in small (T1a,b) invasive breast carcinomas. Cancer Res 1997; 57: 1259–1263.
Barbareschi M, van Tinteren H, Mauri FA, Veronese S, Peterse H, Maisonneuve P et al. p27(kip1) expression in breast carcinomas: an immunohistochemical study on 512 patients with long-term follow-up. Int J Cancer 2000; 89: 236–241.
Reed W, Florems VA, Holm R, Hannisdal E, Nesland JM . Elevated levels of p27, p21 and cyclin D1 correlate with positive oestrogen and progesterone receptor status in node-negative breast carcinoma patients. Virchows Arch 1999; 435: 116–124.
De Paola F, Vecci AM, Granato AM, Liverani M, Monti F, Innoceta AM et al. p27/kip1 expression in normal epithelium, benign and neoplastic breast lesions. J Pathol 2002; 196: 26–31.
Troncone G, Migliaccio I, Caleo A, Palmieri EA, Iaccarino A, Sparano L et al. p27(Kip1) expression and grading of breast cancer diagnosed on cytological samples. Diagn Cytopathol 2004; 30: 375–380.
Banerjee S, Saxena N, Sengupta K, Tawfik O, Mayo MS, Banerjee SK . WISP-2 gene in human breast cancer: estrogen and progesterone inducible expression and regulation of tumor cell proliferation. Neoplasia 2003; 5: 63–73.
Moller MB . P27 in cell cycle control and cancer. Leuk Lymphoma 2000; 39: 19–27.
Chen Y, Robles AI, Martinez LA, Liu F, Gimenez-Conti IB, Conti CJ . Expression of G1 cyclins, cyclin-dependent kinases, and cyclin-dependent kinase inhibitors in androgen-induced prostate proliferation in castrated rats. Cell Growth Differ 1996; 7: 1571–1578.
Kossatz U, Dietrich N, Zender L, Buer J, Manns MP, Malek NP . Skp2-dependent degradation of p27kip1 is essential for cell cycle progression. Genes Dev 2004; 18: 2602–2607.
Nakayama K, Nagahama H, Minamishima YA, Miyake S, Ishida N, Hatakeyama S et al. Skp2-mediated degradation of p27 regulates progression into mitosis. Dev Cell 2004; 6: 661–672.
Nakayama KI, Nakayama K . Regulation of the cell cycle by SCF-type ubiquitin ligases. Semin Cell Dev Biol 2005; 16: 323–333.
Spruck C, Strohmaier H, Watson M, Smith AP, Ryan A, Krek TW et al. A CDK-independent function of mammalian Cks1: targeting of SCF(Skp2) to the CDK inhibitor p27Kip1. Mol Cell 2001; 7: 639–650.
Wei S, Chu PC, Chuang HC, Hung WC, Kulp SK, Chen CS . Targeting the oncogenic E3 ligase Skp2 in prostate and breast cancer cells with a novel energy restriction-mimetic agent. PLoS ONE 2012; 7: e47298.
Carrano AC, Eytan E, Hershko A, Pagano M . SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nat Cell Biol 1999; 1: 193–199.
Nakao T, Geddis AE, Fox NE, Kaushansky K . PI3K/Akt/FOXO3a pathway contributes to thrombopoietin-induced proliferation of primary megakaryocytes in vitro and in vivo via modulation of p27(Kip1). Cell Cycle 2008; 7: 257–266.
Zhang S, Huan W, Wei H, Shi J, Fan J, Zhao J et al. FOXO3a/p27kip1 expression and essential role after acute spinal cord injury in adult rat. J Cell Biochem 2013; 114: 354–365.
Greer EL, Brunet A . FOXO transcription factors at the interface between longevity and tumor suppression. Oncogene 2005; 24: 7410–7425.
Lin K, Dorman JB, Rodan A, Kenyon C . daf-16: An HNF-3/forkhead family member that can function to double the life-span of Caenorhabditis elegans. Science 1997; 278: 1319–1322.
Jun JI, Lau LF . Taking aim at the extracellular matrix: CCN proteins as emerging therapeutic targets. Nat Rev Drug Discov 2011; 10: 945–963.
Russo JW, Castellot JJ . CCN5: biology and pathophysiology. J Cell Commun Signal 2010; 4: 119–130.
Viglietto G, Motti ML, Bruni P, Melillo RM, D'Alessio A, Califano D et al. Cytoplasmic relocalization and inhibition of the cyclin-dependent kinase inhibitor p27(Kip1) by PKB/Akt-mediated phosphorylation in breast cancer. Nat Med 2002; 8: 1136–1144.
Jiang Y, Zhao RC, Verfaillie CM . Abnormal integrin-mediated regulation of chronic myelogenous leukemia CD34+ cell proliferation: BCR/ABL up-regulates the cyclin-dependent kinase inhibitor, p27Kip, which is relocated to the cell cytoplasm and incapable of regulating cdk2 activity. Proc Natl Acad Sci USA 2000; 97: 10538–10543.
Levenson AS, Jordan VC . Transfection of human estrogen receptor (ER) cDNA into ER-negative mammalian cell lines. J Steroid Biochem Mol Biol 1994; 51: 229–239.
Nakayama K, Nagahama H, Minamishima YA, Matsumoto M, Nakamichi I, Kitagawa K et al. Targeted disruption of Skp2 results in accumulation of cyclin E and p27(Kip1), polyploidy and centrosome overduplication. EMBO J 2000; 19: 2069–2081.
Haque I, De A, Majumder M, Mehta S, McGregor D, Banerjee SK et al. The matricellular protein CCN1/Cyr61 is a critical regulator of Sonic Hedgehog in pancreatic carcinogenesis. J Biol Chem 2012; 287: 38569–38579.
Banerjee S, Sengupta K, Saxena NK, Dhar K, Banerjee SK . Epidermal growth factor induces WISP-2/CCN5 expression in estrogen receptor-alpha-positive breast tumor cells through multiple molecular cross-talks. Mol Cancer Res 2005; 3: 151–162.
Banerjee S, Sengupta K, Dhar K, Mehta S, D’Amore PA, Dhar G et al. Breast cancer cells secreted platelet-derived growth factor-induced motility of vascular smooth muscle cells is mediated through neuropilin-1. Mol Carcinog 2006; 45: 871–880.
Dhar K, Banerjee S, Dhar G, Sengupta K, Banerjee SK . Insulin-like growth factor-1 (IGF-1) induces WISP-2/CCN5 via multiple molecular cross-talks and is essential for mitogenic switch by IGF-1 axis in estrogen receptor-positive breast tumor cells. Cancer Res 2007; 67: 1520–1526.
Acknowledgements
These studies were supported by VA Merit Award grants (SKB and SB).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Haque, I., Banerjee, S., De, A. et al. CCN5/WISP-2 promotes growth arrest of triple-negative breast cancer cells through accumulation and trafficking of p27Kip1 via Skp2 and FOXO3a regulation. Oncogene 34, 3152–3163 (2015). https://doi.org/10.1038/onc.2014.250
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2014.250
This article is cited by
-
The role of CCNs in controlling cellular communication in the tumor microenvironment
Journal of Cell Communication and Signaling (2023)
-
WISP2 promotes cell proliferation via targeting ERK and YAP in ovarian cancer cells
Journal of Ovarian Research (2020)
-
The emerging role of WISP proteins in tumorigenesis and cancer therapy
Journal of Translational Medicine (2019)
-
WISP2 exhibits its potential antitumor activity via targeting ERK and E-cadherin pathways in esophageal cancer cells
Journal of Experimental & Clinical Cancer Research (2019)
-
MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway
Cell Death & Disease (2019)