Advertisement

Breast Cancer Research and Treatment

, Volume 93, Issue 2, pp 101–110 | Cite as

RhoC-GTPase is a Novel Tissue Biomarker Associated with Biologically Aggressive Carcinomas of the Breast

  • Celina G. KleerEmail author
  • Kent A. Griffith
  • Michael S. Sabel
  • Gary Gallagher
  • Kenneth L. van  Golen
  • Zhi-Fen Wu
  • Sofia D. Merajver
Report

Summary

Background. There is a need for reliable predictors of breast cancer aggressiveness that will further refine the staging classification and help guide the implementation of novel therapies. We have identified RhoC as being nearly always overexpressed in the most aggressive form of breast cancer, inflammatory breast cancer (IBC); in subsequent work we identified RhoC to be a promising marker of aggressive behavior in breast cancers less than 1 cm in diameter. We hypothesized that RhoC expression would identify aggressive, non-IBC tumors breast cancer patients at any stage with worse outcomes defined as recurrence and/or metastasis.

Methods. We constructed four high-density tissue microarrays (TMAs) using 801 tissue cores from 280 patients. These tissues represent a wide range of normal breast and breast disease, including intraductal hyperplasia, ductal carcinoma in situ (DCIS), invasive carcinomas, and distant metastases. The TMAs were immunostained using a polyclonal anti-RhoC antibody developed in our laboratory. Cytoplasmic RhoC expression was scored as negative, weak, moderate, or strong by a previously validated scoring schema.

Results. RhoC expression increases with breast cancer progression. All samples of normal breast epithelium had negative to weak staining, whereas staining intensity increased in hyperplasia, DCIS, invasive carcinoma, and metastases (Kruskal–Wallis p < 0.001). In patients with invasive carcinoma, high RhoC expression was associated with features of aggressive behavior including high histologic grade, positive lymph nodes, and negative hormonal receptor status. High RhoC expression was a predictor of overall survival in patients with breast cancer (log rank test, p = 0.002) and was associated with 100% increase in the risk of death as compared to patients with low RhoC expression. Importantly, high RhoC was an independent predictor of poor response to doxorubicin-based chemotherapy with a hazard ratio of 3.1 and a 95% CI of 1.2–7.7 (p = 0.02).

Conclusion. RhoC expression increases with breast cancer progression and RhoC protein level in tumor tissue is strongly associated with biologically aggressive invasive carcinomas of the breast. RhoC expression, if validated, may identify patients who are less likely benefit from doxorubicin therapy and suggests RhoC overexpression as a new target for intervention.

Keywords

Breast Cancer Invasive Carcinoma Inflammatory Breast Cancer Breast Disease Breast Cancer Progression 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Jemal, A, Murray, T, Samuels, A, Ghafoor, A, Ward, E, Thun, MJ 2003Cancer statistics CACancer J Clin53526PubMedGoogle Scholar
  2. 2.
    Ellis, M, Hayes, D, Lippman, M 2000Treatment of metastatic diseaseHarris, JLippman, MEMorrow, M eds. Diseases of the BreastLippincott-RavenPhiladelpha749798Google Scholar
  3. 3.
    Hayes, DF, Isaacs, C, Stearns, V 2001Prognostic factors in breast cancer: current and new predictors of metastasisJ Mammary Gland Biol Neoplasia6375392CrossRefPubMedGoogle Scholar
  4. 4.
    Hayes, DF, Trock, B, Harris, AL 1998Assessing the clinical impact of prognostic factors: when is “statistically significant” clinically useful?Breast Cancer Res Treat52305319CrossRefPubMedGoogle Scholar
  5. 5.
    Clark, GM 1996Prognostic and predictive factorsHarris, LM. JMorrow, MHellman, S eds. , Diseases of the BreastLipponcott-Raven PublishersPhiladephia461485Google Scholar
  6. 6.
    Hayes,  2000Do we need prognostic factors in nodal-negative breast cancer?Arbiter Eur J Cancer36302306CrossRefGoogle Scholar
  7. 7.
    Golen, KL, Davies, S, Wu, ZF, Wang, Y, Bucana, CD, Root, H, Chandrasekharappa, S, Strawderman, M, Ethier, SP, Merajver, SD 1999A novel putative low-affinity insulin-like growth factor-binding protein, LIBC (lost in inflammatory breast cancer), and RhoC GTPase correlate with the inflammatory breast cancer phenotypeClin Cancer Res525112519PubMedGoogle Scholar
  8. 8.
    Golen, KL, Wu, ZF, Qiao, XT, Bao, L, Merajver, SD 2000RhoC GTPase overexpression modulates induction of angiogenic factors in breast cellsNeoplasia2418425CrossRefPubMedGoogle Scholar
  9. 9.
    Golen, KL, Wu, ZF, Qiao, XT, Bao, LW, Merajver, SD 2000RhoC GTPase, a novel transforming oncogene for human mammary epithelial cells that partially recapitulates the inflammatory breast cancer phenotypeCancer Res6058325838PubMedGoogle Scholar
  10. 10.
    Kleer, CG, Golen, KL, Zhang, Y, Wu, ZF, Rubin, MA, Merajver, SD 2002Characterization of RhoC expression in benign and malignant breast disease : a potential new marker for small breast carcinomas with metastatic abilityAm J Pathol160579584PubMedGoogle Scholar
  11. 11.
    Kleer, CG, Cao, Q, Varambally, S, Shen, R, Ota, I, Tomlins, SA, Ghosh, D, Sewalt, RG, Otte, AP, Hayes, DF, Sabel, MS, Livant, D, Weiss, SJ, Rubin, MA, Chinnaiyan, AM 2003EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cellsProc Natl Acad Sci USA1001160611611CrossRefPubMedGoogle Scholar
  12. 12.
    Perrone, EE, Theoharis, C, Mucci, NR, Hayasaka, S, Taylor, JM, Cooney, KA, Rubin, MA 2000Tissue microarray assessment of prostate cancer tumor proliferation in African-American and white menJ Natl Cancer Inst92937939CrossRefPubMedGoogle Scholar
  13. 13.
    Elston, EW, Ellis, IO 1993Method for grading breast cancerJ Clin Pathol46189190Google Scholar
  14. 14.
    Manley, S, Mucci, NR, Marzo, AM, Rubin, MA 2001Relational database structure to manage high-density tissue microarray data and images for pathology studies focusing on clinical outcome: the prostate specialized program of research excellence modelAm J Pathol159837843PubMedGoogle Scholar
  15. 15.
    Rhodes, DR, Sanda, MG, Otte, AP, Chinnaiyan, AM, Rubin, MA 2003Multiplex biomarker approach for determining risk of prostate-specific antigen-defined recurrence of prostate cancerJ Natl Cancer Inst95661668PubMedGoogle Scholar
  16. 16.
    Nobes, CD, Hall, A 1995Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodiaCell815362CrossRefPubMedGoogle Scholar
  17. 17.
    Hall, A, Nobes, CD 2000Rho GT Pases molecular switches that control the organization and dynamics of the actin cytoskeletonPhilos Trans R Soc Lond B Biol Sci355965970CrossRefPubMedGoogle Scholar
  18. 18.
    Leung, T, Chen, XQ, Manser, E, Lim, L 1996The p160 RhoA-binding kinase ROK alpha is a member of a kinase family and is involved in the reorganization of the cytoskeletonMol Cell Biol1653135327PubMedGoogle Scholar
  19. 19.
    Kleer, CG, Zhang, Y, Pan, Q, Gallagher, G, Wu, M, Wu, ZF, Merajver, SD 2004WISP3 and RhoC guanosine triphosphatase cooperate in the development of inflammatory breast cancerBreast Cancer Res6R110R115CrossRefGoogle Scholar
  20. 20.
    Wozniak, MA, Desai, R, Solski, PA, Der, CJ, Keely, PJ 2003ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrixJ Cell Biol163583595CrossRefPubMedGoogle Scholar
  21. 21.
    Suwa, H, Ohshio, G, Imamura, T, Watanabe, G, Arii, S, Imamura, M, Narumiya, S, Hiai, H, Fukumoto, M 1998Overexpression of the RhoC gene correlates with progression of ductal adenocarcinoma of the pancreasBr J Cancer77147152PubMedGoogle Scholar
  22. 22.
    Horiuchi, A, Imai, T, Wang, C, Ohira, S, Feng, Y, Nikaido, T, Konishi, I 2003Up-regulation of small GTPases, RhoA and RhoC, is associated with tumor progression in ovarian carcinomaLab Invest83861870PubMedGoogle Scholar
  23. 23.
    Kamai, T, Tsujii, T, Arai, K, Takagi, K, Asami, H, Ito, Y, Oshima, H 2003Significant association of Rho/ROCK pathway with invasion and metastasis of bladder cancerClin Cancer Res926322641PubMedGoogle Scholar
  24. 24.
    Kondo, T, Sentani, K, Oue, N, Yoshida, K, Nakayama, H, Yasui, W 2004Expression of RHOC is associated with metastasis of gastric carcinomasPathobiology711925CrossRefPubMedGoogle Scholar
  25. 25.
    Shikada, Y, Yoshino, I, Okamoto, T, Fukuyama, S, Kameyama, T, Maehara, Y 2003Higher expression of RhoC is related to invasiveness in non-small cell lung carcinomaClin Cancer Res952825286PubMedGoogle Scholar
  26. 26.
    Wang, W, Yang, LY, Yang, ZL, Huang, GW, Lu, WQ 2003Expression and significance of RhoC gene in hepatocellular carcinomaWorld J Gastroenterol919501953PubMedGoogle Scholar
  27. 27.
    Ikoma, T, Takahashi, T, Nagano, S, Li, YM, Ohno, Y, Ando, K, Fujiwara, T, Fujiwara, H, Kosai, K 2004A definitive role of RhoC in metastasis of orthotopic lung cancer in miceClin Cancer Res1011921200PubMedGoogle Scholar
  28. 28.
    Clark, EA, Golub, TR, Lander, ES, Hynes, RO 2000Genomic analysis of metastasis reveals an essential role for RhoCNature406532535CrossRefPubMedGoogle Scholar
  29. 29.
    Carr, KM, Bittner, M, Trent, JM 2003Gene-expression profiling in human cutaneous melanomaOncogene2230763080CrossRefPubMedGoogle Scholar
  30. 30.
    Seabra, MC 1998Membrane association and targeting of prenylated Ras-like GTPasesCell Signal10167172CrossRefPubMedGoogle Scholar
  31. 31.
    Kirschmeier, PT, Whyte, D, Wilson, O, Bishop, WR, Pai, JK 2001In vivo prenylation analysis of Ras and Rho proteinsMethods Enzymol332115127PubMedGoogle Scholar
  32. 32.
    Adamson, P, Marshall, CJ, Hall, A, Tilbrook, PA 1992Post-translational modifications of p21rho proteinsJ Biol Chem2672003320038PubMedGoogle Scholar
  33. 33.
    Sinensky, M 2000Recent advances in the study of prenylated proteinsBiochim Biophys Acta148493106PubMedGoogle Scholar
  34. 34.
    Bishop, AL, Hall, A 2000Rho GTPases and their effector proteinsBiochem J348 241255CrossRefPubMedGoogle Scholar
  35. 35.
    Omer, CA, Chen, Z, Diehl, RE, Conner, MW, Chen, HY, Trumbauer, ME, Gopal-Truter, S, Seeburger, G, Bhimnathwala, H, Abrams, MT, Davide, JP, Ellis, MS, Gibbs, JB, Greenberg, I, Koblan, KS, Kral, AM, Liu, D, Lobell, RB, Miller, PJ, Mosser, SD, O’Neill, TJ, Rands, E, Schaber, MD, Senderak, ET, Oliff, A, Kohl, NE 2000Mouse mammary tumor virus-Ki-rasB transgenic mice develop mammary carcinomas that can be growth-inhibited by a farnesyl:protein transferase inhibitorCancer Res6026802688PubMedGoogle Scholar
  36. 36.
    Prendergast, GC, Khosravi-Far, R, Solski, PA, Kurzawa, H, Lebowitz, PF, CJ,  1995Critical role of Rho in cell transformation by oncogenic RasOncogene1022892296PubMedGoogle Scholar
  37. 37.
    Prendergast, GC, Davide, JP, deSolms, SJ, Giuliani, EA, Graham, SL, Gibbs, JB, Oliff, A, Kohl, NE 1994Farnesyltransferase inhibition causes morphological reversion of ras-transformed cells by a complex mechanism that involves regulation of the actin cytoskeletonMol Cell Biol1441934202PubMedGoogle Scholar
  38. 38.
    Lebowitz, PF, Casey, PJ, Prendergast, GC, Thissen, JA 1997Farnesyltransferase inhibitors alter the prenylation and growth-stimulating function of RhoBJ Biol Chem2721559115594CrossRefPubMedGoogle Scholar
  39. 39.
    Golen, KL, Bao, L, DiVito, MM, Wu, Z, Prendergast, GC, Merajver, SD 2002Reversion of RhoC GTPase-induced inflammatory breast cancer phenotype by treatment with a farnesyl transferase inhibitorMol Cancer Ther1575583PubMedGoogle Scholar
  40. 40.
    Collisson, EA, Kleer, C, Wu, M, De, A, Gambhir, SS, Merajver, SD, Kolodney, MS 2003Atorvastatin prevents RhoC isoprenylation, invasion, and metastasis in human melanoma cellsMol Cancer Ther2941948PubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Celina G. Kleer
    • 1
    • 5
    Email author
  • Kent A. Griffith
    • 2
    • 5
  • Michael S. Sabel
    • 3
    • 5
  • Gary Gallagher
    • 1
  • Kenneth L. van  Golen
    • 4
    • 5
  • Zhi-Fen Wu
    • 4
    • 5
  • Sofia D. Merajver
    • 4
    • 5
  1. 1.Department of Pathology and Internal MedicineUniversity of MichiganAnn ArbourUSA
  2. 2.Department of Biostatistics and Internal MedicineUniversity of MichiganAnn ArbourUSA
  3. 3.Department of Surgery and Internal MedicineUniversity of MichiganAnn ArbourUSA
  4. 4.Division of Hematology/OncologyUniversity of MichiganAnn ArbourUSA
  5. 5.Comprehensive Cancer CenterUniversity of MichiganAnn ArbourUSA

Personalised recommendations