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.
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
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
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
Bast RC, Hennessy B, Mills GB (2009) The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer 9:415–428
Bell DA (2005) Origins and molecular pathology of ovarian cancer. Mod Pathol 18(Suppl 2):S19–S32
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
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
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
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
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
Dickinson RE, Duncan WC (2010) The SLIT–ROBO pathway: a regulator of cell functions with implications for the reproductive system. Reproduction 139:697–704
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
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
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
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
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
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
Hinck L (2004) The versatile roles of ‘axon guidance’ cues in tissue morphogenesis. Dev Cell 7:783–793
Hohenester E (2008) Structural insight into Slit–Robo signalling. Biochem Soc Trans 36:251–256
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
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
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
Kolch W (2000) Meaningful relationships: the regulationg of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J 351:289–305
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
Legg JA, Herbert JM, Clissold P, Bicknell R (2008) Slits and roundabouts in cancer, tumour angiogenesis and endothelial cell migration. Angiogenesis 11:13–21
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
Liao WX, Wing DA, Geng JG, Chen DB (2010b) Perspectives of SLIT/ROBO signaling in placental angiogenesis. Histol Histopathol 25:1181–1190
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
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
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
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
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
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
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
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
Neri D, Bicknell R (2005) Tumour vascular targeting. Nat Rev Cancer 5:436–446
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
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
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
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
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
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
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
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
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
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
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
Ypsilanti AR, Zagar Y, Chedotal A (2010) Moving away from the midline: new developments for Slit and Robo. Development 137:1939–1952
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
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
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
Corresponding author
Rights 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
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-011-0806-2