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PLCγ contributes to metastasis of in situ-occurring mammary and prostate tumors

Oncogene volume 26, pages 3020–3026 (2007)Cite this article

Abstract

Phospholipase C-γ (PLCγ) has been implicated in tumor cell motility required for invasiveness and metastasis. Diminished tumor dissemination has been demonstrated in xenograft models, but studies in naturally-occurring tumors are lacking, having been limited by the timing of the interventions. Therefore, we generated mice that express a doxycycline (DOX)-inducible dominant-negative fragment of PLCγ, PLCz; this approach avoids the in utero lethality caused by the absence of PLCγ. As we targeted two de novo-occurring carcinomas of the mammary (MMTV-driven polyoma middle T antigen model, PyVmT) and prostate (TRAMP model) glands, we limited expression to these epithelial cells by driving DOX transactivator from the prostatein C3 promoter. This avoids the confounding variable of potentially abrogating motility in stromal and endothelial cells. These mice developed normally in the presence of DOX, except for limited mammary development if treated before 6 weeks and immaturity of the prostate gland if treated before 2 weeks of age. DOX-mediated induction of PLCz from age 8 to 16 weeks in PyVmT mice decreased the number of lung metastases by >10-fold (P<0.06) without a detectable effect on in situ tumor cell proliferation or tumor size. Lung metastases were also significantly decreased in the TRAMP model in which the mice expressed the PLCz fragment (P<0.05). DOX treatment itself had no effect on tumor size or metastasis in control mice, nor did it affect tumor dissemination in nontransgenic littermates. In conclusion, abrogation of the PLCγ signaling pathway can limit the metastatic potential of carcinomas.

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References

  • Bodnar RJ, Yates CC, Wells A . (2006). IP-10 blocks vascular endothelial growth factor-induced endothelial cell motility and tube formation via inhibition of calpain. Circ Res 98: 617–625.

    Article  CAS  Google Scholar 

  • Bornfeldt KE, Raines EW, Nakano T, Graves LM, Krebs EG, Ross R . (1994). Insulin-like growth factor-I and platelet-derived growth factor-BB induce directed migration of human arterial smooth muscle cells via signaling pathways that are distinct from those of proliferation. J Clin Invest 93: 1266–1274.

    Article  CAS  Google Scholar 

  • Carpenter G . (2000). The EGF receptor: a nexus for trafficking and signaling. Bioessays 22: 697–707.

    Article  CAS  Google Scholar 

  • Carpenter G, Ji Q . (1999). Phospholipase C-gamma as a signal-transducing element. Exp Cell Res 253: 15–24.

    Article  CAS  Google Scholar 

  • Chen P, Gupta K, Wells A . (1994a). Cell movement elicited by epidermal growth factor receptor requires kinase and autophosphorylation but is separable from mitogenesis. J Cell Biol 124: 547–555.

    Article  CAS  Google Scholar 

  • Chen P, Xie H, Sekar MC, Gupta K, Wells A . (1994b). Epidermal growth factor receptor-mediated cell motility: phospholipase C activity is required, but mitogen-activated protein kinase activity is not sufficient for induced cell movement. J Cell Biol 127: 847–857.

    Article  CAS  Google Scholar 

  • Chen P, Xie H, Wells A . (1996). Mitogenic signaling from the egf receptor is attenuated by a phospholipase C-gamma/protein kinase C feedback mechanism. Mol Biol Cell 7: 871–881.

    Article  CAS  Google Scholar 

  • Claessens F, Celis L, Peeters B, Heyns W, Verhoeven G, Rombauts W . (1989). Functional characterization of an androgen response element in the first intron of the C3(1) gene of prostatic binding protein. Biochem Biophys Res Commun 164: 833–840.

    Article  CAS  Google Scholar 

  • Dittman T, Husemann A, Schewe Y, Hofer JR, Niggemann B, Zanker KS et al. (2002). Induction of cancer cell migration by epidermal growth factor is initiated by specific phosphorylation of tyrosine 1248 of c-erbB-2 receptor via EGFR. FASEB J 16: 1823–1825.

    Article  Google Scholar 

  • Fidler IJ . (2003). The pathogenesis of cancer metastasis: the ‘seed and soil’ hypothesis revisited. Nat Rev Cancer 3: 453–458.

    Article  CAS  Google Scholar 

  • Gingrich JR, Barrios RJ, Kattan MW, Nahm HS, Finegold MJ, Greenberg NM . (1997). Androgen-independent prostate cancer progression in the TRAMP model. Cancer Res 57: 4687–4691.

    CAS  PubMed  Google Scholar 

  • Gingrich JR, Barrios RJ, Morton RA, Boyce BF, DeMayo FJ, Finegold MJ et al. (1996). Metastatic prostate cancer in a transgenic mouse. Cancer Res 56: 4096–4102.

    CAS  PubMed  Google Scholar 

  • Glading A, Chang P, Lauffenburger DA, Wells A . (2000). Epidermal growth factor receptor activation of calpain is required for fibroblast motility and occurs via an ERK/MAP kinase signaling pathway. J Biol Chem 275: 2390–2398.

    Article  CAS  Google Scholar 

  • Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H . (1995). Transcriptional activation by tetracyclines in mammalian cells. Science 268: 1766–1769.

    Article  CAS  Google Scholar 

  • Guy CT, Webster MA, Schaller M, Parsons TJ, Cardiff RD, Muller WJ . (1992). Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. Proc Natl Acad Sci USA 89: 10578–10582.

    Article  CAS  Google Scholar 

  • Hofmann A, Nolan GP, Blau HM . (1996). Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette. Proc Natl Acad Sci USA 93: 5185–5190.

    Article  CAS  Google Scholar 

  • Homma Y, Takenawa T . (1992). Inhibitory effect of src homology (SH) 2/SH3 fragments of phospholipase C-gamma on the catalytic activity of phospholipase C isoforms. Identification of a novel phospholipase C inhibitor region. J Biol Chem 267: 21844–21849.

    CAS  PubMed  Google Scholar 

  • Iwabu A, Smith K, Allen FD, Lauffenburger DA, Wells A . (2004). Epidermal growth factor induces fibroblast contractility and motility via a protein kinase C delta-dependent pathway. J Biol Chem 279: 14551–14560.

    Article  CAS  Google Scholar 

  • Jakobsson PA, Eneroth CM, Killander D, Moberger G, Martensson B . (1973). Histologic classification and grading of malignancy in carcinoma of the larynx. Acta Radiol Ther Phys Biol 12: 1–8.

    Article  CAS  Google Scholar 

  • Ji Q-S, Ermini S, Baulida J, Sun F-L, Carpenter G . (1998a). Epidermal growth factor signaling and mitogenesis in Plcg1 null mouse embryonic fibroblasts. Mol Biol Cell 9: 749–757.

    Article  CAS  Google Scholar 

  • Ji QS, Ermini S, Baulida J, Sun FL, Carpenter G . (1998b). Epidermal growth factor signaling and mitogenesis in Plcg1 null mouse embryonic fibroblasts. Mol Biol Cell 9: 749–757.

    Article  CAS  Google Scholar 

  • Jones NP, Peak J, Brader S, Eccles SA, Katan M . (2005). PLCgamma1 is essential for early events in integrin signalling required for cell motility. J Cell Sci 118: 2695–2706.

    Article  CAS  Google Scholar 

  • Kassis J, Lauffenburger DA, Turner T, Wells A . (2001). Tumor invasion as dysregulated cell motility. Semin Cancer Biol 11: 105–117.

    Article  CAS  Google Scholar 

  • Kassis J, Moellinger J, Lo H, Greenberg NM, Kim HG, Wells A . (1999). A role for phospholipase C-gamma-mediated signaling in tumor cell invasion. Clin Cancer Res 5: 2251–2260.

    CAS  PubMed  Google Scholar 

  • Kundra V, Escobedo JA, Kazlauskas A, Kim HK, Rhee SG, Williams LT et al. (1994). Regulation of chemotaxis by the platelet-derived growth factor receptor-beta. Nature 367: 474–476.

    Article  CAS  Google Scholar 

  • Maroulakou IG, Anver M, Garrett L, Green JE . (1994). Prostate and mammary adenocarcinoma in transgenic mice carrying a rat C3(1) simian virus 40 large tumor antigen fusion gene. Proc Natl Acad Sci USA 91: 11236–11240.

    Article  CAS  Google Scholar 

  • Mouneimne G, Soon L, DesMarais V, Sidani M, Song X, Yip SC et al. (2004). Phospholipase C and cofilin are required for carcinoma cell directionality in response to EGF stimulation. J Cell Biol 166: 697–708.

    Article  CAS  Google Scholar 

  • Muraoka RS, Dumont N, Ritter CA, Dugger TC, Brantley DM, Chen J et al. (2002). Blockade of TGF-beta inhibits mammary tumor cell viability, migration, and metastases. J Clin Invest 109: 1551–1559.

    Article  CAS  Google Scholar 

  • Price JT, Tiganis T, Agarwal A, Djakiew D, Thompson EW . (1999). Epidermal growth factor promotes MDA-MB-231 breast cancer cell migration through a phosphatidylinositol 3′-kinase and phospholipase C-dependent mechanism. Cancer Res 59: 5475–5478.

    CAS  PubMed  Google Scholar 

  • Reddy CC, Wells A, Lauffenburger DA . (1996). Receptor-mediated effects on ligand availability influence relative mitogenic potencies of epidermal growth factor and transforming growth factor alpha. J Cell Physiol 166: 512–522.

    Article  CAS  Google Scholar 

  • Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G et al. (2003). Cell migration: integrating signals from front to back. Science 302: 1704–1709.

    Article  CAS  Google Scholar 

  • Tehranian A, Morris DW, Min BH, Bird DJ, Cardiff RD, Barry PA . (1996). Neoplastic transformation of prostatic and urogenital epithelium by the polyoma virus middle T gene. Am J Pathol 149: 1177–1191.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Thomas SM, Coppelli FM, Song JI, Kassis J, Gooding W, Wells A et al. (2002). Epidermal growth factor receptor-stimulated activation of phospholipase Cy-1 and its role in invasion of head and neck squamous cell carcinoma in vitro. Proc Am Assoc Cancer Res 43: 542–543.

    Google Scholar 

  • Turner T, Epps-Fung MV, Kassis J, Wells A . (1997). Molecular inhibition of phospholipase cgamma signaling abrogates DU-145 prostate tumor cell invasion. Clin Cancer Res 3: 2275–2282.

    CAS  PubMed  Google Scholar 

  • Wang W, Goswami S, Sahai E, Wyckoff JB, Segall JE, Condeelis JS . (2005). Tumor cells caught in the act of invading: their strategy for enhanced cell motility. Trends Cell Biol 15: 138–145.

    Article  CAS  Google Scholar 

  • Webster MA, Martin-Soudant N, Nepveu A, Cardiff RD, Muller WJ . (1998). The induction of uterine leiomyomas and mammary tumors in transgenic mice expressing polyomavirus (PyV) large T (LT) antigen is associated with the ability of PyV LT antigen to form specific complexes with retinoblastoma and CUTL1 family members. Oncogene 16: 1963–1972.

    Article  CAS  Google Scholar 

  • Wells A . (2000). Tumor invasion: role of growth factor-induced cell motility. Adv Cancer Res 78: 31–101.

    Article  CAS  Google Scholar 

  • Wells A, Kassis J, Solava J, Turner T, Lauffenburger DA . (2002). Growth factor-induced cell motility in tumor invasion. Acta Oncologica 41: 124–130.

    Article  CAS  Google Scholar 

  • Xie H, Turner T, Wang MH, Singh RK, Siegal GP, Wells A . (1995). In vitro invasiveness of DU-145 human prostate carcinoma cells is modulated by EGF receptor-mediated signals. Clin Exp Metast 13: 407–419.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Norman Greenberg for the gift of the TRAMP mice and Dr William Muller for the gift of the MMTV-PyV mice. Dr Peter Barry kindly provided the prostatein promoter construct. This work was supported in part by grants from the Department of Defense Congressionally Mandated Medical Research Program on Prostate and Breast Cancer and a VA Merit Award.

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Author notes

  1. J Kassis

    Present address: Laboratory for Pathology, NCI/NIH, Bethesda, MD, 20892, USA

  2. H G Kim

    Present address: Comprehensive Cancer Center, University of Alabama Birmingham, Birmingham, AL, 35294, USA

Authors and Affiliations

  1. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    C R Shepard, J Kassis & A Wells

  2. Department of Pathology, Veterans Affairs Medical Center Pittsburgh, Pittsburgh, PA, USA

    D L Whaley

  3. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    H G Kim

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  1. C R Shepard
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Correspondence to A Wells.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Shepard, C., Kassis, J., Whaley, D. et al. PLCγ contributes to metastasis of in situ-occurring mammary and prostate tumors. Oncogene 26, 3020–3026 (2007). https://doi.org/10.1038/sj.onc.1210115

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