Skip to main content
Log in

Expression and roles of Slit/Robo in human ovarian cancer

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

The Slit glycoproteins and their Roundabout (Robo) receptors regulate migration and growth of many types of cells including human cancer cells. However, little is known about the expression and roles of Slit/Robo in human ovarian cancer. Herein, we examined the expression of Slit/Robo in human normal and malignant ovarian tissues and its potential participation in regulating migration and proliferation of human ovarian cancer cells using two ovarian cancer cell lines, OVCAR-3 and SKOV-3. We demonstrated that Slit2/3 and Robo1 were immunolocalized primarily in stromal cells in human normal ovaries and in cancer cells in many histotypes of ovarian cancer tissues. Protein expression of Slit2/3 and Robo1/4 was also identified in OVCAR-3 and SKOV-3 cells. However, recombinant human Slit2 did not significantly affect SKOV-3 cell migration, and OVCAR-3 and SKOV-3 cell proliferation. Slit2 also did not induce ERK1/2 and AKT1 phosphorylation in OVCAR-3 and SKOV-3 cells. The current findings indicate that three major members (Slit2/3 and Robo1) of Slit/Robo family are widely expressed in the human normal and malignant ovarian tissues and in OVCAR-3 and SKOV-3 cells. However, Slit/Robo signaling may not play an important role in regulating human ovarian cancer cell proliferation and migration.

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

Access this article

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

Instant access to the full article PDF.

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

Similar content being viewed by others

References

  • American Cancer Society at http://www.cancer.org: Cancer Facts & Figures 2010

  • Andrews WD, Barber M, Parnavelas JG (2007) Slit–Robo interactions during cortical development. J Anat 211:188–198

    Article  PubMed  Google Scholar 

  • Ayhan A, Kurman RJ, Yemelyanova A, Vang R, Logani S, Seidman JD, ShihIe M (2009) Defining the cut point between low-grade and high-grade ovarian serous carcinomas: a clinicopathologic and molecular genetic analysis. Am J Surg Pathol 33:1220–1224

    Article  PubMed  Google Scholar 

  • Ba-Charvet KTN, Brose K, Ma L, Wang KH, Marillat V, Sotelo C, Tessier-Lavigne M, Chédotal A (2001) Diversity and specificity of actions of Slit2 proteolytic fragments in axon guidance. J Neurosci 21:4281–4289

    CAS  Google Scholar 

  • Bast RC, Hennessy B, Mills GB (2009) The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer 9:415–428

    Article  PubMed  CAS  Google Scholar 

  • Bell DA (2005) Origins and molecular pathology of ovarian cancer. Mod Pathol 18(Suppl 2):S19–S32

    Article  PubMed  CAS  Google Scholar 

  • Brose K, Bland KS, Wang KH, Arnott D, Henzel W, Goodman CS, Tessier-Lavigne M, Kidd T (1999) Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell 96:795–806

    Article  PubMed  CAS  Google Scholar 

  • Chédotal A, Kerjan G, Moreau-Fauvarque C (2005) The brain within the tumor: new roles for axon guidance molecules in cancers. Cell Death Differ 12:1044–1056

    Article  PubMed  Google Scholar 

  • Dallol A, Da Silva NF, Viacava P, Minna JD, Bieche I, Maher ER, Latif F (2002a) SLIT2, a human homologue of the Drosophila Slit2 gene, has tumor suppressor activity and is frequently inactivated in lung and breast cancers. Cancer Res 62:5874–5880

    PubMed  CAS  Google Scholar 

  • Dallol A, Forgacs E, Martinez A, Sekido Y, Walker R, Kishida T, Rabbitts P, Masher ER, Minna JD, Latif F (2002b) Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. Oncogene 21:3020–3028

    Article  PubMed  CAS  Google Scholar 

  • Dallol A, Morton D, Masher ER, Latif F (2003) SLIT2 axon guidance molecule is frequently inactivated in colorectal cancer and suppresses growth of colorectal carcinoma cells. Cancer Res 63:1054–1058

    PubMed  CAS  Google Scholar 

  • Dickinson RE, Duncan WC (2010) The SLIT–ROBO pathway: a regulator of cell functions with implications for the reproductive system. Reproduction 139:697–704

    Article  PubMed  CAS  Google Scholar 

  • Dickinson RE, Dallol A, Bieche I, Krex D, Morton D, Maher ER, Latif F (2004) Epigenetic inactivation of SLIT3 and SLIT1 genes in human cancers. Br J Cancer 91:2071–2078

    Article  PubMed  CAS  Google Scholar 

  • Dickinson RE, Myers M, Duncan WC (2008) Novel regulated expression of the SLIT/ROBO pathway in the ovary: possible role during luteolysis in women. Endocrinology 149:5024–5034

    Article  PubMed  CAS  Google Scholar 

  • Dickinson RE, Hryhorskyj L, Tremewan H, Hogg K, Thomson AA, McNeilly AS, Duncan WC (2010) Involvement of the SLIT/ROBO pathway in follicle development in the fetal ovary. Reproduction 139:395–407

    Article  PubMed  CAS  Google Scholar 

  • Duncan WC, McDonald SE, Dickinson RE, Shaw JLV, Lourenco PC, Wheelhouse N, Lee KF, Critchley HO, Horne AW (2010) Expression of the repulsive SLIT/ROBO pathway in the human endometrium and fallopian tube. Mol Hum Reprod 16:950–959

    Article  PubMed  CAS  Google Scholar 

  • Grieshammer U, Le M, Plump AS, Wang F, Tessier-Lavigne M, Martin GR (2004) SLIT2-mediated ROBO2 signaling restricts kidney induction to a single site. Dev cell 6:709–717

    Article  PubMed  CAS  Google Scholar 

  • Grone J, Doebler O, Loddenkemper C, Hotz B, Buhr HJ, Bhargava S (2006) Robo1/Robo4: differential expression of angiogenic markers in colorectal cancer. Oncol Rep 15:1437–1443

    PubMed  Google Scholar 

  • Hinck L (2004) The versatile roles of ‘axon guidance’ cues in tissue morphogenesis. Dev Cell 7:783–793

    Article  PubMed  CAS  Google Scholar 

  • Hohenester E (2008) Structural insight into Slit–Robo signalling. Biochem Soc Trans 36:251–256

    Article  PubMed  CAS  Google Scholar 

  • Ito H, Funahashi S, Yamauchi N, Shibahara J, Midorikawa Y, Kawai S, Kinoshita Y, Watanabe A, Hippo Y, Ohtomo T, Iwanari H, Nakajima A, Makuuchi M, Fukayama M, Hirata Y, Hamakubo T, Kodama T, Tsuchiya M, Aburatani H (2006) Identification of ROBO1 as a novel hepatocellular carcinoma antigen and a potential therapeutic and diagnostic target. Clin Cancer Res 12:3257–3264

    Article  PubMed  CAS  Google Scholar 

  • Jiang YZ, Wang K, Fang R, Zheng J (2010) Expression of aryl hydrocarbon receptor in human placentas and fetal tissues. J Histochem Cytochem 58:679–685

    Article  PubMed  CAS  Google Scholar 

  • Jones CA, London NR, Chen H, Park KW, Sauvaget D, Stockton RA, Wythe JD, Suh W, Larrieu-Lahargue F, Mukouyama YS, Lindblom P, Seth P, Frias A, Nishiya N, Ginsberg MH, Gerhardt H, Zhang K, Li DY (2008) Robo4 stabilizes the vascular network by inhibiting pathologic angiogenesis and endothelial hyperpermeability. Nat Med 14:448–453

    Article  PubMed  CAS  Google Scholar 

  • Kolch W (2000) Meaningful relationships: the regulationg of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J 351:289–305

    Article  PubMed  CAS  Google Scholar 

  • Latil A, Chene L, Cochant-Priollet B, Mangin P, Fournier G, Berthon P, Cussenot O (2003) Quantification of expression of netrins, slits and their receptors in human prostate tumors. Int J Cancer 103:306–315

    Article  PubMed  CAS  Google Scholar 

  • Legg JA, Herbert JM, Clissold P, Bicknell R (2008) Slits and roundabouts in cancer, tumour angiogenesis and endothelial cell migration. Angiogenesis 11:13–21

    Article  PubMed  Google Scholar 

  • Liao WX, Feng L, Zheng J, Chen DB (2010a) Deciphering mechanisms controlling placental artery endothelial cell migration stimulated by vascular endothelial growth factor. Endocrinology 151:3432–3444

    Article  PubMed  CAS  Google Scholar 

  • Liao WX, Wing DA, Geng JG, Chen DB (2010b) Perspectives of SLIT/ROBO signaling in placental angiogenesis. Histol Histopathol 25:1181–1190

    PubMed  CAS  Google Scholar 

  • Little MH, Wilkinson L, Brown DL, Piper M, Yamada T, Stow JL (2001) Dual trafficking of Slit3 to mitochondria and cell surface demonstrates novel localization for Slit protein. Am J Physiol Cell Physiol 281:C486–C495

    PubMed  CAS  Google Scholar 

  • Liu J, Zhang L, Wang D, Shen H, Jiang M, Mei P, Hayden PS, Sedor JR, Hu H (2003) Congenital diaphragmatic hernia, kidney agenesis and cardiac defects associated with Slit3-deficiency in mice. Mech Dev 120:1059–1070

    Article  PubMed  CAS  Google Scholar 

  • Ma S, Liu X, Geng JG, Guo SW (2010) Increased SLIT immunoreactivity as a biomarker for recurrence in endometrial carcinoma. Am J Obstet Gynecol 202:68.e1–68.e11

    Google Scholar 

  • Malpica A, Deavers MT, Lu K, Bodurka DC, Atkinson EN, Gershenson DM, Silva EG (2004) Grading ovarian serous carcinoma using a two-tier system. Am J Surg Pathol 28:496–504

    Article  PubMed  Google Scholar 

  • Mertsch S, Schmitz N, Jeibmann A, Geng JG, Paulus W, Senner V (2008) Slit2 involvement in glioma cell migration is mediated by Robo1 receptor. J Neurooncol 87:1–7

    Article  PubMed  CAS  Google Scholar 

  • Morlot C, Thielens NM, Ravelli RB, Hemrika W, Romijn RA, Gros P, Cusack S, McCarthy AA (2007) Structural insights into the Slit–Robo complex. Proc Natl Acad Sci USA 104:14923–14928

    Google Scholar 

  • Morris MR, Hesson LB, Wagner KJ, Morgan NV, Astuti D, Lees RD, Cooper WN, Lee J, Gentle D, Macdonald F, Kishida T, Grundy R, Yao M, Latif F, Maher ER (2003) Multigene methylation analysis of Wilms’ tumour and adult renal cell carcinoma. Oncogene 22:6794–6801

    Article  PubMed  CAS  Google Scholar 

  • Narayan G, Goparaju C, Arias-Pulido H, Kaufmann AM, Schneider A, Dürst M, Mansukhani M, Pothuri B, Murty VV (2006) Promoter hypermethylation-mediated inactivation of multiple Slit–Robo pathway genes in cervical cancer progression. Mol Cancer 5:16

    Article  PubMed  Google Scholar 

  • Neri D, Bicknell R (2005) Tumour vascular targeting. Nat Rev Cancer 5:436–446

    Article  PubMed  CAS  Google Scholar 

  • Patel K, Nash JA, Itoh A, Liu Z, Sundaresan V, Pini A (2001) Slit proteins are not dominant chemorepellents for olfactory tract and spinal motor axons. Development 128:5031–5037

    PubMed  CAS  Google Scholar 

  • Piper M, Anderson R, Dwivedy A, Weinl C, van Horck F, Leung KM, Cogill E, Holt C (2006) Signaling mechanisms underlying Slit2-induced collapse of Xenopus retinal growth cones. Neuron 49:215–228

    Article  PubMed  CAS  Google Scholar 

  • Plump AS, Erskine L, Sabatier C, Brose K, Epstein CJ, Goodman CS, Mason CA, Tessier-Lavigne M (2002) Slit1 and Slit2 cooperate to prevent premature midline crossing of retinal axons in the mouse visual system. Neuron 33:219–232

    Article  PubMed  CAS  Google Scholar 

  • Seki M, Watanabe A, Enomoto S, Kawamura T, Ito H, Kodama T, Hamakubo T, Aburatani H (2010) Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells. FEBS Lett 584:2909–2915

    Article  PubMed  CAS  Google Scholar 

  • Seth P, Lin Y, Hanai J, Shivalingappa V, Duyao MP, Sukhatme VP (2005) Magic roundabout, a tumor endothelial marker: expression and signaling. Biochem Biophys Res Commun 332:533–541

    Article  PubMed  CAS  Google Scholar 

  • Sundaresan V, Chung G, Heppell-Parton A, Xiong J, Grundy C, Roberts I, James L, Cahn A, Bench A, Douglas J, Minna J, Sekido Y, Lerman M, Latif F, Bergh J, Li H, Lowe N, Ogilvie D, Rabbitts P (1998) Homozygous deletions at 3p12 in breast and lung cancer. Oncogene 17:1723–1729

    Article  PubMed  CAS  Google Scholar 

  • Tanno T, Tanaka Y, Sugiura T, Akiyoshi H, Takenaka S, Kuwamura M, Yamate J, Ohashi F, Kubo K, Tsuyama S (2006) Expression patterns of the slit subfamily mRNA in canine malignant mammary tumors. J Vet Med Sci 68:1173–1177

    Article  PubMed  CAS  Google Scholar 

  • Wang B, Xiao Y, Ding BB, Zhang N, Yuan X, Gui L, Qian KX, Duan S, Chen Z, Rao Y, Geng JG (2003) Induction of tumor angiogenesis by Slit–Robo signaling and inhibition of cancer growth by blocking Robo activity. Cancer Cell 4:19–29

    Article  PubMed  Google Scholar 

  • Wang K, Song Y, Chen DB, Zheng J (2008) Protein phosphatase 3 differentially modulates vascular endothelial growth factor and fibroblast growth factor 2-stimulated cell proliferation and signaling in ovine fetoplacental artery endothelial cells. Biol Reprod 79:704–710

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Chan DW, Liu VW, Chiu P, Ngan HY (2010) Differential functions of growth factor receptor-bound protein 7 (GRB7) and its variant GRB7v in ovarian carcinogenesis. Clin Cancer Res 16:2529–2539

    Article  PubMed  CAS  Google Scholar 

  • Xian J, Clark KJ, Fordham R, Pannell R, Rabbitts TH, Rabbitts PH (2001) Inadequate lung development and bronchial hyperplasia in mice with a targeted deletion in the Dutt1/Robo1 gene. Proc Natl Acad Sci USA 98:15062–15066

    Article  PubMed  CAS  Google Scholar 

  • Ypsilanti AR, Zagar Y, Chedotal A (2010) Moving away from the midline: new developments for Slit and Robo. Development 137:1939–1952

    Article  PubMed  CAS  Google Scholar 

  • Yuan W, Rao Y, Babiuk RP, Greer JJ, Wu JY, Ornitz DM (2003) A genetic model for a central (septum transversum) congenital diaphragmatic hernia in mice lacking Slit3. Proc Natl Acad Sci USA 100:5217–5222

    Article  PubMed  CAS  Google Scholar 

  • Zheng J, Bird IM, Melsaether AN, Magness RR (1999) Activation of the mitogen-activated protein kinase cascade is necessary but not sufficient for basic fibroblast growth factor- and epidermal growth factor-stimulated expression of endothelial nitric oxide synthase in ovine fetoplacental artery endothelial cells. Endocrinology 140:1399–1407

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported in part by the National Institutes of Health grants HL64703 and HD38843 (JZ).

Conflict of interest

The authors have declared that no competing interests exist.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jing Zheng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dai, C.F., Jiang, Y.Z., Li, Y. et al. Expression and roles of Slit/Robo in human ovarian cancer. Histochem Cell Biol 135, 475–485 (2011). https://doi.org/10.1007/s00418-011-0806-2

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-011-0806-2

Keywords

Navigation