Journal Home > Volume 5 , Issue 1
Journal of Neurorestoratology 2017, 5(1): 143-153 https://doi.org/10.2147/JN.S134363
Original Research | Open Access | Issue | Published: 18 July 2017

Effects of IGFBP-2 on proliferation and differentiation in neural stem cell line C17.2

Show Author's Information Yujia Deng1,  Lei Wang1,2,  Lite Ge1,3,  Da Duan1,  Yi Zhuo1,  Ting Yuan1,  Weiping Yan1,  Peiqi Huang1,  Xiaohua Teng1( ),  Ming Lu1,3( )
Department of Neurosurgery, The Second Affiliated Hospital of Hunan Normal University (163 Hospital of the People’s Liberation Army), Changsha,
Department of Neurosurgery, Affiliated Haikou Hospital, Xiangya School of Central South University, Haikou,
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha, People’s Republic of China
Keywords:
proliferation, differentiation, IGFBP-2, C17.2 cells
Cite this article:
Deng Y, Wang L, Ge L, et al. Effects of IGFBP-2 on proliferation and differentiation in neural stem cell line C17.2. Journal of Neurorestoratology, 2017, 5(1): 143-153. https://doi.org/10.2147/JN.S134363
Download citation
EndNote(RIS)
BibTeX

455

Views

13

Downloads

Citations

2

Crossref

3

WoS

0

Scopus

N/A

CSCD

Abstract Full text About this article
Objective:

Insulin-like growth factor binding protein-2 (IGFBP-2), a member of a highly conserved family of six insulin-like growth factor binding proteins (IGFBPs), can regulate several cellular processes through IGF-dependent or IGF-independent pathway. Recent studies have provided solid evidence for the importance to delineate that olfactory ensheathing cells (OEC)-conditioned medium (OCM) can not only facilitate the differentiation of neural stem cell line (C17.2) into neurons, but also promote the survival and proliferation. We have previously reported that IGFBP-2 was detected in OCM. This study is designed to investigate the roles of IGFBP-2 for the regulation of C17.2 differentiation and proliferation.

Methods and results:

IGFBP-2 was identified and upregulated in OCM to compare with astrocytes-conditioned medium by shotgun proteomics and semiquantitative proteomic analysis. In order to investigate whether exogenous IGFBP-2 could stimulate proliferation in C17.2 cells and differentiate it into glia or neuron, we used various concentrations of IGFBP-2 to induce C17.2 cells which were cultured in DMEM/F12. The results showed that exogenous IGFBP-2 can promote proliferation in C17.2 cells, but had little effect on differentiation. Interestingly, we also found that IGFBP-2 could induce C17.2 cells to differentiate into astrocytes, while inhibiting their differentiation into neurons in a dose-dependent manner when cultured C17.2 cells in OCM. Changes in cell morphology were imaged under a light microscope, and proliferating cells were counted. Cell viability was determined by MTT. In addition, Western blot, immunofluorescence, and flow cytometry analysis were performed to detect protein expression patterns of proliferation-related antigen, proliferating cell nuclear antigen, neuroectodermal stem cell marker, neuron specific class III beta tubulin, and glial fibrillary acidic protein.

Conclusion:

Exogenous IGFBP-2 could stimulate proliferation in C17.2 cells, and promote the differentiation of C17.2 cells into astrocytes induced by OCM. Its mechanism is related to activation of the extracellular signal-regulated kinase 1/2 pathway.


menu
Abstract
Full text
Outline
About this article

Effects of IGFBP-2 on proliferation and differentiation in neural stem cell line C17.2

Show Author's information Yujia Deng1, Lei Wang1,2, Lite Ge1,3, Da Duan1, Yi Zhuo1, Ting Yuan1, Weiping Yan1, Peiqi Huang1, Xiaohua Teng1( ), Ming Lu1,3( )
Department of Neurosurgery, The Second Affiliated Hospital of Hunan Normal University (163 Hospital of the People’s Liberation Army), Changsha,
Department of Neurosurgery, Affiliated Haikou Hospital, Xiangya School of Central South University, Haikou,
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha, People’s Republic of China

Abstract

Objective:

Insulin-like growth factor binding protein-2 (IGFBP-2), a member of a highly conserved family of six insulin-like growth factor binding proteins (IGFBPs), can regulate several cellular processes through IGF-dependent or IGF-independent pathway. Recent studies have provided solid evidence for the importance to delineate that olfactory ensheathing cells (OEC)-conditioned medium (OCM) can not only facilitate the differentiation of neural stem cell line (C17.2) into neurons, but also promote the survival and proliferation. We have previously reported that IGFBP-2 was detected in OCM. This study is designed to investigate the roles of IGFBP-2 for the regulation of C17.2 differentiation and proliferation.

Methods and results:

IGFBP-2 was identified and upregulated in OCM to compare with astrocytes-conditioned medium by shotgun proteomics and semiquantitative proteomic analysis. In order to investigate whether exogenous IGFBP-2 could stimulate proliferation in C17.2 cells and differentiate it into glia or neuron, we used various concentrations of IGFBP-2 to induce C17.2 cells which were cultured in DMEM/F12. The results showed that exogenous IGFBP-2 can promote proliferation in C17.2 cells, but had little effect on differentiation. Interestingly, we also found that IGFBP-2 could induce C17.2 cells to differentiate into astrocytes, while inhibiting their differentiation into neurons in a dose-dependent manner when cultured C17.2 cells in OCM. Changes in cell morphology were imaged under a light microscope, and proliferating cells were counted. Cell viability was determined by MTT. In addition, Western blot, immunofluorescence, and flow cytometry analysis were performed to detect protein expression patterns of proliferation-related antigen, proliferating cell nuclear antigen, neuroectodermal stem cell marker, neuron specific class III beta tubulin, and glial fibrillary acidic protein.

Conclusion:

Exogenous IGFBP-2 could stimulate proliferation in C17.2 cells, and promote the differentiation of C17.2 cells into astrocytes induced by OCM. Its mechanism is related to activation of the extracellular signal-regulated kinase 1/2 pathway.

Keywords: proliferation, differentiation, IGFBP-2, C17.2 cells

References(45)

1.
Carpentier PA, Palmer TD. Immune influence on adult neural stem cell regulation and function. Neuron. 2009;64(1):79–92.
DOI Google Scholar
2.
Lipson AC, Widenfalk J, Lindqvist E, Ebendal T, Olson L. Neurotrophic properties of olfactory ensheathing glia. Exp Neurol. 2003;180(2):167–171.
DOI Google Scholar
3.
Ao Q, Wang AJ, Chen GQ, Wang SJ, Zuo HC, Zhang XF. Combined transplantation of neural stem cells and olfactory ensheathing cells for the repair of spinal cord injuries. Med Hypotheses. 2007;69(6):1234–1237.
DOI Google Scholar
4.
Liu Y, Teng X, Yang X, et al. Shotgun proteomics and network analysis between plasma membrane and extracellular matrix proteins from rat olfactory ensheathing cells. Cell Transplant. 2010;19(2):133–146.
DOI Google Scholar
5.
Ranke MB, Elmlinger M. Functional role of insulin-like growth factor binding proteins. Horm Res. 1997;48(Suppl 4):9–15.
DOI Google Scholar
6.
Firth SM, Baxter RC. Cellular actions of the insulin-like growth factor binding proteins. Endocr Rev. 2002;23(6):824–854.
DOI Google Scholar
7.
Russo VC, Gluckman PD, Feldman EL, Werther GA. The insulin-like growth factor system and its pleiotropic functions in brain. Endocr Rev. 2005;26(7):916–943.
DOI Google Scholar
8.
Jones JI, Clemmons DR. Insulin-like growth factors and their binding proteins: biological actions. Endocr Rev. 1995;16(1):3–34.
DOI Google Scholar
9.
Suzuki Y, Takeda M. Expression of insulin-like growth factor family in the rat olfactory epithelium. Anat Embryol (Berl). 2002;205(5–6):401–405.
DOI Google Scholar
10.
Federico G, Maremmani C, Cinquanta L, Baroncelli GI, Fattori B, Saggese G. Mucus of the human olfactory epithelium contains the insulin-like growth factor-I system which is altered in some neurodegenerative diseases. Brain Res. 1999;835(2):306–314.
DOI Google Scholar
11.
Kiepe D, Ulinski T, Powell DR, Durham SK, Mehls O, Tönshoff B. Differential effects of insulin-like growth factor binding proteins-1, -2, -3, and -6 on cultured growth plate chondrocytes. Kidney Int. 2002;62(5):1591–1600.
DOI Google Scholar
12.
Hoeflich A, Reisinger R, Lahm H, et al. Insulin-like growth factor-binding protein 2 in tumorigenesis: protector or promoter. Cancer Res. 2001;61(24):8601–8610.
Google Scholar
13.
Park SH, Kim KW, Kim JC. The Role of Insulin-Like Growth Factor Binding Protein 2 (IGFBP2) in the regulation of corneal fibroblast differentiation. Invest Ophthalmol Vis Sci. 2015;56(12):7293–7302.
DOI Google Scholar
14.
Bartling B, Koch A, Simm A, Scheubel R, Silber RE, Santos AN. Insulin-like growth factor binding proteins-2 and -4 enhance the migration of human CD34-/CD133+ hematopoietic stem and progenitor cells. Int J Mol Med. 2010;25(1):89–96.
DOI Google Scholar
15.
Migita T, Narita T, Asaka R, et al. Role of insulin-like growth factor binding protein 2 in lung adenocarcinoma: IGF-independent antiapoptotic effect via caspase-3. Am J Pathol. 2010;176(4):1756–1766.
DOI Google Scholar
16.
Huynh H, Zheng J, Umikawa M, et al. IGF binding protein 2 supports the survival and cycling of hematopoietic stem cells. Blood. 2011;118(12):3236–3243.
DOI Google Scholar
17.
Kim DS, Cho HJ, Yang SK, Shin JW, Huh CH, Park KC. Insulin-like growth factor-binding protein contributes to the proliferation of less proliferative cells in forming skin equivalents. Tissue Eng Part A. 2009;15(5):1075–1080.
DOI Google Scholar
18.
Pereira JJ, Meyer T, Docherty SE, et al. Bimolecular interaction of insulin-like growth factor (IGF) binding protein-2 with alphavbeta3 negatively modulates IGF-I-mediated migration and tumor growth. Cancer Res. 2004;64(3):977–984.
DOI Google Scholar
19.
Hsieh D, Hsieh A, Stea B, Ellsworth R. IGFBP2 promotes glioma tumor stem cell expansion and survival. Biochem Biophys Res Commun. 2010;397(2):367–372.
DOI Google Scholar
20.
Lin Y, Jiang T, Zhou K, et al. Plasma IGFBP-2 levels predict clinical outcomes of patients with high-grade gliomas. Neuro Oncol. 2009;11(5):468–476.
DOI Google Scholar
21.
Klempt ND, Klempt M, Gunn AJ, Singh K, Gluckman PD. Expression of insulin-like growth factor-binding protein 2 (IGF-BP 2) following transient hypoxia-ischemia in the infant rat brain. Brain Res Mol Brain Res. 1992;15(1–2):55–61.
DOI Google Scholar
22.
Fletcher L, Isgor E, Sprague S, et al. Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury. BMC Neurosci. 2013;14:158.
DOI Google Scholar
23.
Taylor RM, Snyder EY. Widespread engraftment of neural progenitor and stem-like cells throughout the mouse brain. Transplant Proc. 1997;29(1–2):845–847.
DOI Google Scholar
24.
Gerdes J, Schwab U, Lemke H, Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer. 1983;31(1):13–20.
DOI Google Scholar
25.
Lenczewski A, Terlikowski S, Sulkowska M, et al. [Expression of PCNA antigen in cellular proliferative activity of cervical carcinoma]. Ginekol Pol. 2000;71(9):1164–1168. Polish.
Google Scholar
26.
Maga G, Hubscher U. Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J Cell Sci. 2003;116(Pt 15):3051–3060.
DOI Google Scholar
27.
Duan D, Rong M, Zeng Y, et al. Electrophysiological characterization of NSCs after differentiation induced by OEC conditioned medium. Acta Neurochir (Wien). 2011;153(10):2085–2090.
DOI Google Scholar
28.
Brandt K, Grünler J, Brismar K, Wang J. Effects of IGFBP-1 and IGFBP-2 and their fragments on migration and IGF-induced proliferation of human dermal fibroblasts. Growth Horm IGF Res. 2015;25(1):34–40.
DOI Google Scholar
29.
Celebi B, Mantovani D, Pineault N. Insulin-like growth factor binding protein-2 and neurotrophin 3 synergize together to promote the expansion of hematopoietic cells ex vivo. Cytokine. 2012;58(3):327–331.
DOI Google Scholar
30.
Shi LH, Zhu XQ, Zhao GH, Xia YB, Zhang YS. Expression of insulin-like growth factor binding protein-2 in gastric carcinoma and its relationship with cell proliferation. World J Gastroenterol. 2006;12(39):6285–6289.
DOI Google Scholar
31.
Zhang L, Huang W, Chen J, Zhou X, Lu Z, Zhou H. Expression of IGFBP2 in gastric carcinoma and relationship with clinicopathologic parameters and cell proliferation. Dig Dis Sci. 2007;52(1):248–253.
DOI Google Scholar
32.
Russo VC, Schütt BS, Andaloro E, et al. Insulin-like growth factor binding protein-2 binding to extracellular matrix plays a critical role in neuroblastoma cell proliferation, migration, and invasion. Endocrinology. 2005;146(10):4445–4455.
DOI Google Scholar
33.
Kiyama S, Morrison K, Zellweger T, et al. Castration-induced increases in insulin-like growth factor-binding protein 2 promotes proliferation of androgen-independent human prostate LNCaP tumors. Cancer Res. 2003;63(13):3575–3584.
Google Scholar
34.
Frommer KW, Reichenmiller K, Schutt BS, et al. IGF-independent effects of IGFBP-2 on the human breast cancer cell line Hs578T. J Mol Endocrinol. 2006;37(1):13–23.
DOI Google Scholar
35.
Xi G, Wai C, DeMambro V, Rosen CJ, Clemmons DR. IGFBP-2 directly stimulates osteoblast differentiation. J Bone Miner Res. 2014;29(11):2427–2438.
DOI Google Scholar
36.
Zhang J, Wang B, Xiao Z, et al. Olfactory ensheathing cells promote proliferation and inhibit neuronal differentiation of neural progenitor cells through activation of Notch signaling. Neuroscience. 2008;153(2):406–413.
DOI Google Scholar
37.
Wang L, Jiang M, Duan D, et al. Hyperthermia-conditioned OECs serum-free-conditioned medium induce NSC differentiation into neuron more efficiently by the upregulation of HIF-1 alpha and binding activity. Transplantation. 2014;97(12):1225–1232.
DOI Google Scholar
38.
Wood AW, Schlueter PJ, Duan C. Targeted knockdown of insulin-like growth factor binding protein-2 disrupts cardiovascular development in zebrafish embryos. Mol Endocrinol .2005;19(4):1024–1034.
DOI Google Scholar
39.
Alkharobi H, Alhodhodi A, Hawsawi Y, et al. IGFBP-2 and -3 co-ordinately regulate IGF1 induced matrix mineralisation of differentiating human dental pulp cells. Stem Cell Res. 2016;17(3):517–522.
DOI Google Scholar
40.
Xi G, Wai C, DeMambro V, Rosen CJ, Clemmons DR. IGFBP-2 directly stimulates osteoblast differentiation. J Bone Miner Res. 2014;29(11):2427–2438.
DOI Google Scholar
41.
Huat TJ, Khan AA, Pati S, Mustafa Z, Abdullah JM, Jaafar H. IGF-1 enhances cell proliferation and survival during early differentiation of mesenchymal stem cells to neural progenitor-like cells. BMC Neurosci. 2014;15:91.
DOI Google Scholar
42.
Zhao L, Feng Y, Chen X, et al. Effects of IGF-1 on neural differentiation of human umbilical cord derived mesenchymal stem cells. Life Sciences. 2016,151:93.
DOI Google Scholar
43.
Zhang X, Zhang L, Cheng X, et al. IGF-1 promotes Brn-4 expression and neuronal differentiation of neural stem cells via the PI3K/Akt pathway. PLoS One. 2014;9(12):e113801.
DOI Google Scholar
44.
Mendes KN, Wang GK, Fuller GN, Zhang W. JNK mediates insulin-like growth factor binding protein 2/integrin alpha5-dependent glioma cell migration. Int J Oncol. 2010;37(1):143–153.
DOI Google Scholar
45.
Han S, Li Z, Master LM, Wu A. Exogenous IGFBP-2 promotes proliferation, invasion, and chemoresistance to temozolomide in glioma cells via the integrin β1-ERK pathway. Br J Cancer. 2014;111(7):1400–1409.
DOI Google Scholar
Publication history
Copyright
Acknowledgements
Rights and permissions

Publication history

Published: 18 July 2017
Issue date: December 2017

Copyright

© 2017 The Author(s).

Acknowledgements

This work was supported by National Natural Science Foundation of China (No. 81371358) and Scientific Research Fund of Heilongjiang Provincial Education Department (15C0832). The authors would like to thank Miao Jiang from Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education in the College of Life Sciences, Hunan Normal University for his assistance.

Rights and permissions

© 2017 Deng et al. This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).

Return