Role of autophagy on bone marrow mesenchymal stem‑cell proliferation and differentiation into neurons

Li,Bo; Duan,Ping; Li,Caifang; Jing,Ying; Han,Xuefei; Yan,Wenhai; Xing,Ying

doi:10.3892/mmr.2015.4673

Role of autophagy on bone marrow mesenchymal stem‑cell proliferation and differentiation into neurons

Authors:
- Bo Li
- Ping Duan
- Caifang Li
- Ying Jing
- Xuefei Han
- Wenhai Yan
- Ying Xing
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Affiliations: Department of Physiology, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Department of Pathophysiology, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Stem Cell Research Center of Basic Medicine College, Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: December 10, 2015 https://doi.org/10.3892/mmr.2015.4673
Pages: 1413-1419

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Abstract

The purpose of the present study was to investigate the role of autophagy on rat bone marrow mesenchymal stem cell (BMSC) proliferation, apoptosis and differentiation into neurons. After treatment with rapamycin, 3‑methyladenine (3‑MA) or chloroquine, the cell cycle, apoptosis, expression of neuron‑specific enolase (NSE) and the mean fluorescence intensity (MFI) of Notch1 in BMSCs were examined by flow cytometry. The expression of microtubule‑associated protein 2 (MAP2), Notch1 and Hes1 was investigated by western blot analysis. The results showed that after induction of autophagy using rapamycin, the proliferation of BMSCs was inhibited. Furthermore, the S‑phase population was significantly decreased compared to that in the control group (P<0.05). In addition, the percentage of NSE‑positive cells and the expression of MAP2 were significantly increased compared to those in the control group (P<0.05). The MFI of Notch1 was markedly upregulated compared to that in the control group (P<0.05). When autophagy was inhibited by 3‑MA or chloroquine, the percentage of apoptotic cells and NSE‑positive cells as well as the expression of MAP2 were markedly reduced compared to those in the control group (P<0.05). Furthermore, western blot analysis showed that Notch1 and Hes1 were decreased in the rapamycin‑treated group, while they were not affected by 3‑MA or chloroquine. The present study indicated that induction of autophagy in BMSCs decreased their S‑phase population, promoted their differentiation into neurons and promoted the expression of NSE and MAP2. The mechanisms underlying this process may be linked to the regulation of autophagy‑induced inhibition of the Notch1 signaling pathway.

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1	Lapierre LR, Gelino S, Meléndez A and Hansen M: Autophagy and lipid metabolism coordinately modulate life span in germline-less C. elegans. Curr Biol. 21:1507–1514. 2011. View Article : Google Scholar : PubMed/NCBI
2	Zeng M and Zhou JN: Roles of autophagy and mTOR signaling in neuronal differentiation of mouse neuroblastoma cells. Cell Signal. 20:659–665. 2008. View Article : Google Scholar : PubMed/NCBI
3	Greenwald I and Kovall R: Notch signaling: Genetics and structure. WormBook. 17:1–28. 2013. View Article : Google Scholar
4	Chiba S: Notch signaling in stem cell systems. Stem cells. 24:2437–2447. 2006. View Article : Google Scholar : PubMed/NCBI
5	Bray SJ: Notch signalling: A simple pathway becomes complex. Nat Rev Mol Cell Biol. 7:678–689. 2006. View Article : Google Scholar : PubMed/NCBI
6	Louvi A and Artavanis-Tsakonas S: Notch and disease: A growing field. Semin Cell Dev Biol. 23:473–480. 2012. View Article : Google Scholar : PubMed/NCBI
7	Chen BY, Wang X, Chen LW and Luo ZJ: Molecular targeting regulation of proliferation and differentiation of the bone marrow-derived mesenchymal stem cells or mesenchymal stromal cells. Curr Drug Targets. 13:561–571. 2012. View Article : Google Scholar : PubMed/NCBI
8	Woodbury D, Schwarz EJ, Prockop DJ and Black IB: Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res. 61:364–370. 2000. View Article : Google Scholar : PubMed/NCBI
9	Zhang R, Li J and Xie J: Efficient in vitro labeling rabbit bone marrow-derived mesenchymal stem cells with SPIO and differentiating into neural-like cells. Mol Cells. 37:650–655. 2014. View Article : Google Scholar : PubMed/NCBI
10	Caplan AI: Why are MSCs therapeutic? New data: New insight. J Pathol. 217:318–324. 2009. View Article : Google Scholar
11	Anbari F, Khalili MA, Bahrami AR, Khoradmehr A, Sadeghian F, Fesahat F and Nabi A: Intravenous transplantation of bone marrow mesenchymal stem cells promotes neural regeneration after traumatic brain injury. Neural Regen Res. 9:919–923. 2014. View Article : Google Scholar : PubMed/NCBI
12	Jing L and Jia Y, Lu J, Han R, Li J, Wang S, Peng T and Jia Y: MicroRNA-9 promotes differentiation of mouse bone mesenchymal stem cells into neurons by Notch signaling. Neuroreport. 22:206–211. 2011. View Article : Google Scholar : PubMed/NCBI
13	Hayashi T, Wakao S, Kitada M, Ose T, Watabe H, Kuroda Y, Mitsunaga K, Matsuse D, Shigemoto T, Ito A, et al: Autologous mesenchymal stem cell-derived dopaminergic neurons function in parkinsonian macaques. J Clin Invest. 123:272–284. 2013. View Article : Google Scholar :
14	Todd LR, Gomathinayagam R and Sankar U: A novel Gfer-Drp1 link in preserving mitochondrial dynamics and function in pluripotent stem cells. Autophagy. 6:821–822. 2010. View Article : Google Scholar : PubMed/NCBI
15	Mukhopadhyay S, Panda PK, Sinha N, Das DN and Bhutia SK: Autophagy and apoptosis: Where do they meet? Apoptosis. 19:555–566. 2014. View Article : Google Scholar : PubMed/NCBI
16	Meléndez A and Neufeld TP: The cell biology of autophagy in metazoans: A developing story. Development. 135:2347–2360. 2008. View Article : Google Scholar : PubMed/NCBI
17	Mizushima N, Levine B, Cuervo AM and Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 451:1069–1075. 2008. View Article : Google Scholar : PubMed/NCBI
18	Yang Z and Klionsky DJ: Mammalian autophagy: Core molecular machinery and signaling regulation. Curr Opin Cell Biol. 22:124–131. 2010. View Article : Google Scholar :
19	Yorimitsu T and Klionsky DJ: Autophagy: Molecular machinery for self-eating. Cell Death Differ. 12(Suppl 2): S1542–S1552. 2005. View Article : Google Scholar
20	Guan JL, Simon AK, Prescott M, Menendez JA, Liu F, Wang F, Wang C, Wolvetang E, Vazquez-Martin A and Zhang J: Autophagy in stem cells. Autophagy. 9:830–849. 2013. View Article : Google Scholar : PubMed/NCBI
21	Zirin J and Perrimon N: Drosophila as a model system to study autophagy. Semin Immunopathol. 32:363–372. 2010. View Article : Google Scholar : PubMed/NCBI
22	Rubinsztein DC, Gestwicki JE, Murphy LO and Klionsky DJ: Potential therapeutic applications of autophagy. Nat Rev Drug Discov. 6:304–312. 2007. View Article : Google Scholar : PubMed/NCBI
23	Coller HA, Sang L and Roberts JM: A new description of cellular quiescence. PLoS Biol. 4:e832006. View Article : Google Scholar : PubMed/NCBI
24	Phadwal K, Watson AS and Simon AK: Tightrope act: Autophagy in stem cell renewal, differentiation, proliferation and aging. Cell Mol Life Sci. 70:89–103. 2013. View Article : Google Scholar :
25	Mortensen M, Watson AS and Simon AK: Lack of autophagy in the hematopoietic system leads to loss of hematopoietic stem cell function and dysregulated myeloid proliferation. Autophagy. 7:1069–1070. 2011. View Article : Google Scholar : PubMed/NCBI
26	Kabeya Y, Mizushima N, Yamamoto A, Oshitani-Okamoto S, Ohsumi Y and Yoshimori T: LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci. 117:2805–2812. 2004. View Article : Google Scholar : PubMed/NCBI
27	Komatsu M, Kurokawa H, Waguri S, Taguchi K, Kobayashi A, Ichimura Y, Sou YS, Ueno I, Sakamoto A, Tong KI, et al: The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol. 12:213–223. 2010.PubMed/NCBI
28	Martelli AM, Evangelisti C, Chiarini F, Grimaldi C, Cappellini A, Ognibene A and McCubrey JA: The emerging role of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin signaling network in normal myelopoiesis and leukemogenesis. Biochim Biophys Acta. 1803:991–1002. 2010. View Article : Google Scholar : PubMed/NCBI
29	Wu YT, Tan HL, Shui G, Bauvy C, Huang Q, Wenk MR, Ong CN, Codogno P and Shen HM: Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class I and III phosphoinositide 3-kinase. J Biol Chem. 285:10850–10861. 2010. View Article : Google Scholar : PubMed/NCBI
30	Yoon YH, Cho KS, Hwang JJ, Lee SJ, Choi JA and Koh JY: Induction of lysosomal dilatation, arrested autophagy and cell death by chloroquine in cultured ARPE-19 cells. Invest Ophthalmol Vis Sci. 51:6030–6037. 2010. View Article : Google Scholar : PubMed/NCBI
31	Pantovic A, Krstic A, Janjetovic K, Kocic J, Harhaji-Trajkovic L, Bugarski D and Trajkovic V: Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells. Bone. 52:524–531. 2013. View Article : Google Scholar
32	Lee Y, Jung J, Cho KJ, Lee SK, Park JW, Oh IH and Kim GJ: Increased SCF/c-kit by hypoxia promotes autophagy of human placental chorionic plate-derived mesenchymal stem cells via regulating the phosphorylation of mTOR. J Cell Biochem. 114:79–88. 2013. View Article : Google Scholar
33	Wu J, Niu J, Li X, Li Y, Wang X, Lin J and Zhang F: Hypoxia induces autophagy of bone marrow-derived mesenchymal stem cells via activation of ERK1/2. Cell Physiol Biochem. 33:1467–1474. 2014. View Article : Google Scholar : PubMed/NCBI
34	Li N, Zhang Q, Qian HY, Jin C, Yang Y and Gao R: Atorvastatin induces autophagy of mesenchymal stem cells under hypoxia and serum deprivation conditions by activating the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway. Chin Med J (Engl). 127:1046–1051. 2014.
35	Song C, Song C and Tong F: Autophagy induction is a survival response against oxidative stress in bone marrow-derived mesenchymal stromal cells. Cytotherapy. 26:1361–1370. 2014. View Article : Google Scholar
36	Wang L, Hu X, Zhu W, Jiang Z, Zhou Y, Chen P and Wang J: Increased leptin by hypoxic-preconditioning promotes autophagy of mesenchymal stem cells and protects them from apoptosis. Sci China Life Sci. 57:171–180. 2014. View Article : Google Scholar : PubMed/NCBI
37	Ugland H, Naderi S, Brech A, Collas P and Blomhoff HK: CAMP induces autophagy via a novel pathway involving ERK, cyclin E and Beclin 1. Autophagy. 7:1199–1211. 2011. View Article : Google Scholar : PubMed/NCBI
38	Shintani T and Klionsky DJ: Autophagy in health and disease: A double-edged sword. Science. 306:990–995. 2004. View Article : Google Scholar : PubMed/NCBI
39	Singh R, Xiang Y, Wang Y, Baikati K, Cuervo AM, Luu YK, Tang Y, Pessin JE, Schwartz GJ and Czaja MJ: Autophagy regulates adipose mass and differentiation in mice. J Clin Invest. 119:3329–3339. 2009.PubMed/NCBI
40	Li Y, Wang C, Zhang G, Wang X, Duan R, Gao H, Peng T, Teng J and Jia Y: Role of autophagy and mTOR signaling in neural differentiation of bone marrow mesenchymal stem cells. Cell Biol Int. 38:1337–1343. 2014. View Article : Google Scholar : PubMed/NCBI
41	Patel PN, Yu XM, Jaskula-Sztul R and Chen H: Hesperetin activates the Notch1 signaling cascade, causes apoptosis, and induces cellular differentiation in anaplastic thyroid cancer. Ann Surg Oncol. 21(Suppl 4): S497–S504. 2014. View Article : Google Scholar : PubMed/NCBI
42	Su BH, Qu J, Song M, Huang XY, Hu XM, Xie J, Zhao Y, Ding LC, She L, Chen J, et al: NOTCH1 signaling contributes to cell growth, anti-apoptosis and metastasis in salivary adenoid cystic carcinoma. Oncotarget. 5:6885–6895. 2014. View Article : Google Scholar : PubMed/NCBI
43	Zhao C, Guo H, Li J, Myint T, Pittman W, Yang L, Zhong W, Schwartz RJ, Schwarz JJ, et al: Numb family proteins are essential for cardiac morphogenesis and progenitor differentiation. Development. 141:281–295. 2014. View Article : Google Scholar :
44	Xing Y, Bai RY, Yan WH, Han XF, Duan P, Xu Y and Fan ZG: Expression changes of Notch-related genes during the differentiation of human mesenchymal stem cells into neurons. Sheng Li Xue Bao. 59:267–272. 2007.In Chinese. PubMed/NCBI
45	Barth JM and Köhler K: How to take autophagy and endocytosis up a notch. Biomed Res Int. 2014:9608032014. View Article : Google Scholar : PubMed/NCBI
46	Kwon MH, Callaway H, Zhong J and Yedvobnick B: A targeted genetic modifier screen links the SWI2/SNF2 protein domino to growth and autophagy genes in Drosophila melanogaster. G3 (Bethesda). 3:815–825. 2013. View Article : Google Scholar
47	Yamamoto S, Charng WL and Bellen HJ: Endocytosis and intracellular trafficking of Notch and its ligands. Curr Top Dev Biol. 92:165–200. 2010. View Article : Google Scholar : PubMed/NCBI