Robo3A and Robo3B expression is regulated via alternative promoters and mRNA stability

Cancer Cell International - Tập 16 - Trang 1-10 - 2016
Anke Ruedel1, Mandy Schott1, Thomas Schubert2, Anja Katrin Bosserhoff1
1Institute of Biochemistry, Emil-Fischer-Zentrum, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany
2Institute of Pathology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany

Tóm tắt

The transmembrane receptor family Roundabout (Robo) was described to have an essential role in the developing nervous system. Recent studies demonstrated that Robo3 shows an altered expression in rheumatoid arthritis as well as in melanoma. Until today no detailed studies of the two Robo3 isoforms (Robo3A and Robo3B) and their roles in rheumatoid arthritis synovial fibroblasts, respectively malignant melanoma are available. To get a better understanding regarding the role of Robo3A and Robo3B in the molecular process of rheumatoid arthritis and melanoma the exact characterization of expression and regulation is object of this study. mRNA and protein expression of the transcriptional variants were analyzed by quantitative RT-PCR respectively western blotting and revealed particularly enhanced expression of Robo3B in rheumatoid arthritis and melanoma. Promoter assays and inhibitor studies also disclosed that there is apparently a cell- and isoform-specific regulation of the Robo3 expression. Finally, dissimilar mRNA stabilities of Robo3A and Robo3B are identified as decisive posttranscriptional gene expression control. In summary, this study supported an isotype specific role of Robo3B in disease hinting to different functional roles of each isoform.

Tài liệu tham khảo

Sabatier C, Plump AS, Le M, et al. The divergent Robo family protein rig-1/Robo3 is a negative regulator of slit responsiveness required for midline crossing by commissural axons. Cell. 2004;117(2):157–69. Ballard MS, Hinck L. A roundabout way to cancer. Adv Cancer Res. 2012;114:187–235. Schubert T, Denk AE, Ruedel A, et al. Fragments of SLIT3 inhibit cellular migration. Int J Mol Med. 2012;30(5):1133–7. Dickinson RE, Hryhorskyj L, Tremewan H, et al. Involvement of the SLIT/ROBO pathway in follicle development in the fetal ovary. Reproduction. 2010;139(2):395–407. Ypsilanti AR, Zagar Y, Chedotal A. Moving away from the midline: new developments for slit and Robo. Development. 2010;137(12):1939–52. Zelina P, Blockus H, Zagar Y, et al. Signaling switch of the axon guidance receptor Robo3 during vertebrate evolution. Neuron. 2014;84(6):1258–72. Camurri L, Mambetisaeva E, Davies D, Parnavelas J, Sundaresan V, Andrews W. Evidence for the existence of two Robo3 isoforms with divergent biochemical properties. Mol Cell Neurosci. 2005;30(4):485–93. Challa AK, McWhorter ML, Wang C, Seeger MA, Beattie CE. Robo3 isoforms have distinct roles during zebrafish development. Mech Dev. 2005;122(10):1073–86. Denk AE, Kaufmann S, Stark K, et al. Slit3 inhibits Robo3-induced invasion of synovial fibroblasts in rheumatoid arthritis. Arthr Res Ther. 2010;12(2):R45. Denk AE, Braig S, Schubert T, Bosserhoff AK. Slit3 inhibits activator protein 1-mediated migration of malignant melanoma cells. Int J Mol Med. 2011;28(5):721–6. Ruedel A, Dietrich P, Schubert T, Hofmeister S, Hellerbrand C, Bosserhoff AK. Expression and function of microRNA-188-5p in activated rheumatoid arthritis synovial fibroblasts. Int J Clin Exp Pathol. 2015;8(5):4953–62. Miller LE, Jüsten HP, Schölmerich J, Straub RH. The loss of sympathetic nerve fibers in the synovial tissue of patients with rheumatoid arthritis is accompanied by increased norepinephrine release from synovial macrophages. FASEB J. 2000;14(13):2097–107. Ruedel A, Hofmeister S, Bosserhoff AK. Development of a model system to analyze chondrogenic differentiation of mesenchymal stem cells. Int J Clin Exp Pathol. 2013;6(12):3042–8. Bosserhoff AK, Hofmeister S, Ruedel A, Schubert T. DCC is expressed in a CD166-positive subpopulation of chondrocytes in human osteoarthritic cartilage and modulates CRE activity. Int J Clin Exp Pathol. 2014;7(5):1947–56. Shim J, Karin M. The control of mRNA stability in response to extracellular stimuli. Mol Cells. 2002;14(3):323–31. Gara RK, Kumari S, Ganju A, Yallapu MM, Jaggi M, Chauhan SC. Slit/Robo pathway: a promising therapeutic target for cancer. Drug Discov Today. 2015;20(1):156–64. Han S, Cao C, Tang T, et al. ROBO3 promotes growth and metastasis of pancreatic carcinoma. Cancer Lett. 2015;366(1):61–70. Narayan G, Goparaju C, Arias-Pulido H, et al. Promoter hypermethylation-mediated inactivation of multiple Slit-Robo pathway genes in cervical cancer progression. Mol Cancer. 2006;5:16. Kuphal S, Bosserhoff AK. Influence of the cytoplasmic domain of E-cadherin on endogenous N-cadherin expression in malignant melanoma. Oncogene. 2006;25(2):248–59. Jen JC, Chan WM, Bosley TM, et al. Mutations in a human ROBO gene disrupt hindbrain axon pathway crossing and morphogenesis. Science. 2004;304(5676):1509–13. Singer GA, Wu J, Yan P, Plass C, Huang TH, Davuluri RV. Genome-wide analysis of alternative promoters of human genes using a custom promoter tiling array. BMC Genom. 2008;9:349. Landry JR, Mager DL, Wilhelm BT. Complex controls: the role of alternative promoters in mammalian genomes. Trends Genetics TIG. 2003;19(11):640–8. Nural HF, Todd Farmer W, Mastick GS. The Slit receptor Robo1 is predominantly expressed via the Dutt1 alternative promoter in pioneer neurons in the embryonic mouse brain and spinal cord. Gene Expr Patterns GEP. 2007;7(8):837–45. Thalhamer T, McGrath MA, Harnett MM. MAPKs and their relevance to arthritis and inflammation. Rheumatol (Oxford). 2008;47(4):409–14. Kuphal S, Bosserhoff A. Recent progress in understanding the pathology of malignant melanoma. J Pathol. 2009;219(4):400–9. Shelledy L, Roman D. Vemurafenib: first-in-class BRAF-mutated inhibitor for the treatment of unresectable or metastatic melanoma. J Adv Pract Oncol. 2015;6(4):361–5. Rajagopalan S, Nicolas E, Vivancos V, Berger J, Dickson BJ. Crossing the midline: roles and regulation of Robo receptors. Neuron. 2000;28(3):767–77. Simpson JH, Bland KS, Fetter RD, Goodman CS. Short-range and long-range guidance by Slit and its Robo receptors: a combinatorial code of Robo receptors controls lateral position. Cell. 2000;103(7):1019–32. Chen CY, Shyu AB. AU-rich elements: characterization and importance in mRNA degradation. Trends Biochem Sci. 1995;20(11):465–70. Schiavi SC, Belasco JG, Greenberg ME. Regulation of proto-oncogene mRNA stability. Biochim Biophys Acta. 1992;1114(2–3):95–106. Chen CY, Gherzi R, Andersen JS, et al. Nucleolin and YB-1 are required for JNK-mediated interleukin-2 mRNA stabilization during T-cell activation. Genes Dev. 2000;14(10):1236–48.