Skip to main content

Advertisement

SpringerLink
Log in

R-Ras promotes metastasis of cervical cancer epithelial cells

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Mutations in the small GTPase R-Ras that promote constitutive activation of this signaling molecule have been observed in a variety of invasive cancer cell types. We previously reported that expression of an oncogenic form of R-Ras (R-Ras87L) in a cell line of cervical cancer (C33A cells) augments cell growth in vitro and tumorigenicity in vivo. Because increased tumorigenicity in vivo often precedes metastasis, we now examined whether the expression of R-Ras87L also increased the metastatic potential of C33A cells. Accelerated tumor growth was observed in athymic mice after subcutaneous injection of R-Ras87L-expressing C33A cells. In addition, increased metastasis to the liver, in immunodeficient SCID mice, was observed after intravenous injection of R-Ras87L-expressing C33A cells. Also, R-Ras87L-expressing cells presented decreased membrane expression of MHC class I molecules, and β1 integrins, but increased levels of PI 3-K and Akt activities. C33A cells expressing R-Ras87L also migrated more over collagen I in wound assays. Inhibition of the PI 3-K/Akt/mTOR pathway by pharmacological means blocked R-Ras87L-induced accelerated growth and migration over collagen I. These results suggest oncogenic R-Ras has a central role in cancer progression towards a metastatic phenotype, through the activation of the PI 3-K/Akt/mTOR signaling pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Berrier AL, Mastrangelo AM, Downward J, Ginsberg M, LaFlamme SE (2000) Activated R-Ras, Rac-1, PI 3-kinase and PKCe can each restore cell spreading inhibited by isolated integrin b1 cytoplasmic domains. J Cell Biol 151:1549–1560

    Article  PubMed  CAS  Google Scholar 

  2. Cole AL, Subbanagounder G, Mukhopadhyay S, Berliner JA, Vora DK (2003) Oxidized phospholipid-induced endothelial cell/monocyte interaction is mediated by a cAMP-dependent R-Ras/PI3-kinase pathway. Arterioscler Thromb Vasc Biol 23:1384–1390

    Article  PubMed  CAS  Google Scholar 

  3. Cox AD, Brtva TR, Lowe DG, Der CJ (1994) R-Ras promotes malignant, but not morphologic, transformation of NIH3T3 cells. Oncogene 9:3281–3288

    PubMed  CAS  Google Scholar 

  4. Debnath J, Walker SJ, Brugge JS (2003) Akt activation disrupts mammary acinar architecture and enhances proliferation in an mTOR-dependent manner. J Cell Biol 163:315–326

    Article  PubMed  CAS  Google Scholar 

  5. Felding-Habermann B, O’Toole TE, Smith JW, Fransvea E, Ruggeri ZM, Ginsberg MH, Hughes PE, Pampori N, Shattil SJ, Saven A, Mueller BM (2001) Integrin activation controls metastasis in human breast cancer. Proc Natl Acad Sci USA 98:1853–1888

    Article  PubMed  CAS  Google Scholar 

  6. Gao N, Zhang Z, Jiang B-H, Shi X (2003) Role of PI3K/AKT/mTOR signaling in the cell cycle progression of human prostate cancer. Biochem Biophys Res Commun 310:1124–1132

    Article  PubMed  CAS  Google Scholar 

  7. Garcia-Garcia E, Sanchez-Mejorada G, Rosales C (2001) Phosphatidylinositol 3-kinase and ERK are required for NF-kB activation, but not for phagocytosis. J Leukoc Biol 70:649–658

    PubMed  CAS  Google Scholar 

  8. Ghim SJ, Sundberg J, Delgado G, Jenson AB (2001) The pathogenesis of advanced cervical cancer provides the basis for an empirical therapeutic vaccine. Exp Mol Pathol 71:181–185

    Article  PubMed  CAS  Google Scholar 

  9. Gotoh T, Cai D, Tian X, Feig LA, Lerner A (2000) p130Cas regulates the activity of AND-34, a novel Ral, Rap1, and R-Ras guanine nucleotide exchange factor. J Biol Chem 275:30118–30123

    Article  PubMed  CAS  Google Scholar 

  10. Holly SP, Larson MK, Parise LV (2005) The unique N-terminus of R-Ras is required for Rac activation and precise regulation of cell migration. Mol Biol Cell 16:2458–2469

    Article  PubMed  CAS  Google Scholar 

  11. Hood JD, Cheresh DA (2002) Role of integrins in cell invation and migration. Nat Rev Cancer 2:91–100

    Article  PubMed  Google Scholar 

  12. Huang Y, Rangwala F, Fulkerson PC, Ling B, Reed E, Cox AD, Kamholz J, Ratner N (2004) Role of TC21/R-Ras2 in enhanced migration of neurofibromin-deficient Schwann cells. Oncogene 23:368–378

    Article  PubMed  CAS  Google Scholar 

  13. Huff SY, Quilliam TR, Cox AD, Der CJ (1997) R-Ras is regulated by activators and effectors distinct from those that control Ras function. Oncogene 14:133–143

    Article  PubMed  CAS  Google Scholar 

  14. Jeong H-W, Nam J-O, Kim I-S (2005) The COOH-terminal end of R-Ras alters the motility and morphology of breast epithelial cells through Rho/Rho-kinase. Cancer Res 65:507–515

    PubMed  CAS  Google Scholar 

  15. Juliano RL, Varner JA (1993) Adhesion molecules in cancer: the role of integrins. Curr Opin Cell Biol 5:812–818

    Article  PubMed  CAS  Google Scholar 

  16. Keely PJ, Rusyn EV, Cox AD, Parise LV (1999) R-Ras signals through specific integrin alpha cytoplasmic domains to promote migration and invasion of breast epithelial cells. J Cell Biol 145:1077–1088

    Article  PubMed  CAS  Google Scholar 

  17. Lowe DG, Capon DJ, Delwart E, Sakaguchi AY, Naylor SL, Goeddel DV (1987) Structure of the human and murine R-ras genes, novel genes closely related to ras proto-oncogenes. Cell 48:137–146

    Article  PubMed  CAS  Google Scholar 

  18. Marte BM, Rodriguez-Viciana P, Wennstrom S, Warne PH, Downward J (1996) R-Ras can activate the phosphoinositide 3-kinase but not the MAP kinase arm of the Ras effector pathways. Curr Biol 7:63–70

    Article  Google Scholar 

  19. Maschler S, Wirl G, Spring H, Bredow DV, Sordat I, Beug H, Reichmann E (2005) Tumor cell invasiveness correlates with changes in integrin expression and localization. Oncogene 24:2032–2041

    Article  PubMed  CAS  Google Scholar 

  20. Motoyama S, Ladines-Llave CA, Luis-Villanueva S, Maruo T (2004) The role of human papilloma virus in the molecular biology of cervical carcinogenesis. Kobe J Med Sci 50:9–19

    PubMed  CAS  Google Scholar 

  21. Münger K, Baldwin A, Edwards KM, Hayakawa H, Nguyen CL, Owens M, Grace M, Huh K (2004) Mechanisms of human papillomavirus-induced oncogenesis. J Virol 78:11451–11460

    Article  PubMed  Google Scholar 

  22. Nakada M, Niska JA, Tran NL, McDonough WS, Berens ME (2005) EphB2/R-Ras signaling regulates glioma cell adhesion, growth, and invasion. Am J Pathol 167(2):565–576

    PubMed  CAS  Google Scholar 

  23. Nishigaki M, Aoyagi K, Danjoh I, Fukaya M, Yanagihara K, Sakamoto H, Yoshida T, Sasaki H (2005) Discovery of aberrant expression of R-RAS by cancer-linked DNA hypomethylation in gastric cancer using microarrays. Cancer Res 65:2115–2124

    Article  PubMed  CAS  Google Scholar 

  24. Oinuma I, Ishikawa Y, Katoh H, Negishi M (2004) The Semaphorin 4D receptor Plexin-B1 is a GTPase activating protein for R-Ras. Science 305:862–965

    Article  PubMed  CAS  Google Scholar 

  25. Osada M, Tolkachera T, Li W, Chan TO, Tsichlis PN, Saez R, Kimmelman AC, Chan AM (1999) Differential roles of Akt, Rac, and Raf in R-Ras-mediated cellular transformation adhesion and survival. Mol Cell Biol 9:6333–6344

    Google Scholar 

  26. Parise LV, Weon J, Juliano RL (2000) New aspects of integrin signaling in cancer. Sem Cancer Biol 10:407–414

    Article  CAS  Google Scholar 

  27. Pene F, Claessens YE, Muller O, Viguie F, Mayeux P, Dreyfus F, Lacombe C, Bouscary D (2002) Role of the phosphatidylinositol 3-kinase/Akt and mTOR/P70S6-kinase pathways in the proliferation and apoptosis in multiple myeloma. Oncogene 21:6587–6597

    Article  PubMed  CAS  Google Scholar 

  28. Ponta H, Sleeman J, Herrlich P (1994) Tumor metastasis formation: cell-surface proteins confer metastasis-promoting or -suppressing properties. Biochim Biophys Acta 1198:1–10

    PubMed  CAS  Google Scholar 

  29. Potter JD (1997) Food, nutrition and the prevention of cancer: A global perspective. The American Institute for Cancer Research, Washington DC, p 670

  30. Reyes-Reyes M, Mora N, Gonzalez G, Rosales C (2002) b1 and b2 integrins activate different signalling pathways in monocytes. Biochem J 363:273–280

    Article  PubMed  CAS  Google Scholar 

  31. Reyes-Reyes M, Mora N, Zentella A, Rosales C (2001) Phosphatidylinositol 3-kinase mediates integrin-dependent NF-kB and MAPK activation through separate signaling pathways. J Cell Sci 114:1579–1589

    PubMed  CAS  Google Scholar 

  32. Rincón-Arano H, Rosales R, Mora N, Rodríguez-Castañeda A, Rosales C (2003) R-Ras promotes tumor growth of cervical epithelial cells. Cancer 97:575–585

    Article  PubMed  Google Scholar 

  33. Rodríguez Viciana P, Warne PH, Vanhaesebroeck B, Waterfield MD, Downward J (1996) Activation of phosphoinositide 3-kinase by interaction with Ras and by point mutation. EMBO J 15:2442–2451

    PubMed  Google Scholar 

  34. Saez R, Chan AM-L, Miki T, Aaronson SA (1994) Oncogenic activation of human R-Ras by point mutations analogous to those of prototype H-Ras. Oncogene 9:2977–2982

    PubMed  CAS  Google Scholar 

  35. Schorge JO, Knowles LM, Lea JS (2004) Adenocarcinoma of the cervix. Curr Treat Options Oncol 5:119–127

    PubMed  Google Scholar 

  36. Self AJ, Caron E, Peterson HF, Hall A (2001) Analysis of R-Ras signalling pathways. J Cell Sci 114:1357–1366

    PubMed  CAS  Google Scholar 

  37. Sethi T, Ginsberg MH, Downward J, Hughes PE (1999) The small GTP-binding protein R-Ras can influence integrin activation by antagonizing a Ras/Raf-initiated intetgrin supression pathway. Mol Biol Cell 10:1799–1809

    PubMed  CAS  Google Scholar 

  38. Sirianni MC, Libi F, Campagna M, Rossi D, Capello D, Sciaranghella G, Carbone A, Simonelli C, Monini P, Gaidano G, Ensoli B (2005) Downregulation of the major histocompatibility complex class I molecules by human herpesvirus type 8 and impaired natural killer cell activity in primary effusion lymphoma development. Br J Haematol 130:92–95

    Article  PubMed  CAS  Google Scholar 

  39. Sugatani T, Hruska KA (2005) Akt1/Akt2 and mammalian target of rapamycin/Bim play critical roles in osteoclast differentiation and survival, respectively, whereas Akt is dispensable for cell survival in isolated osteoclast precursors. J Biol Chem 280:3583–3589

    Article  PubMed  CAS  Google Scholar 

  40. Torres Lobaton A, Rojo Herrera G, Torres Rojo A, Hurtado Estrada G, Roman Bassaure E (2004) Cervical cancer. Current view of its epidemiology and risk factors. Ginecol Obstet Mex 72:466–474

    PubMed  Google Scholar 

  41. Woodman CBJ, Collins S, Winter H, Bailey A, Ellis J, Prior P, Yates M, Rollason TP, Young LS (2001) Natural history of cervical human papillomavirus infection in young women: a longitudinal cohort study. Lancet 357:1831–1836

    Article  PubMed  CAS  Google Scholar 

  42. Wozniak MA, Kwong L, Chodniewicz D, Klemke RL, Keely PJ (2005) R-Ras controls membrane protrusion and cell migration through the spatial regulation of Rac and Rho. Mol Biol Cell 16:84–96

    Article  PubMed  CAS  Google Scholar 

  43. Yu Y, Feig LA (2002) Involvement of R-Ras and Ral GTPases in estrogen-independent proliferation of breast cancer cells. Oncogene 21:7557–7568

    Article  PubMed  CAS  Google Scholar 

  44. Zhang Z, Vouri K, Wang HG, Reed JC, Rouslahti E (1996) Integrin activation by R-ras. Cell 85:61–69

    Article  PubMed  CAS  Google Scholar 

  45. Zhu Y, Zhong X, Zheng S, Ge Z, Du Q, Zhang S (2005) Transformation of immortalized colorectal crypt cells by microcystin involving constitutive activation of Akt and MAPK cascade. Carcinogenesis 26:1207–1214

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Adrienne D. Cox for R-Ras constructs, Dr. Julian Downward for the active PI 3-K construct, and Dr. Martin Hemler, and Dr. Eric J. Brown for anti-integrin antibodies. We also thank Dr. Jesus Chimal for helping taking pictures of mouse livers, and Jose Alejandro Marmolejo Valencia for technical assistance. This work was supported by grant 36407-M from Consejo Nacional de Ciencia y Tecnología, Mexico, and by grant IN220703 from DGAPA, Universidad Nacional Autónoma de México, Mexico.

Author information

Authors and Affiliations

  1. Department of Immunology, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Apdo. Postal 70228, Cd. Universitaria, México City, 04510, Mexico

    Nancy Mora & Carlos Rosales

  2. Molecular Biology and Biotechnology, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico City, Mexico

    Ricardo Rosales

Authors
  1. Nancy Mora
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ricardo Rosales
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carlos Rosales
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carlos Rosales.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mora, N., Rosales, R. & Rosales, C. R-Ras promotes metastasis of cervical cancer epithelial cells. Cancer Immunol Immunother 56, 535–544 (2007). https://doi.org/10.1007/s00262-006-0205-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-006-0205-z

Keywords

Search

Navigation