Effects of leukemia inhibitory factor receptor on the adipogenic differentiation of human bone marrow mesenchymal stem cells

Wang,Tao; Yan,Ruiqiao; Xu,Xiaoyuan; Yu,Huan; Wu,Jianfang; Yang,Yaofang; Li,Weidong

doi:10.3892/mmr.2019.10140

Effects of leukemia inhibitory factor receptor on the adipogenic differentiation of human bone marrow mesenchymal stem cells

Authors:
- Tao Wang
- Ruiqiao Yan
- Xiaoyuan Xu
- Huan Yu
- Jianfang Wu
- Yaofang Yang
- Weidong Li
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Affiliations: Key Laboratory of System Bio‑Medicine of Jiangxi Province, Jiujiang University, Jiujiang, Jiangxi 332000, P.R. China, Clinical Skills Center, Affiliated Hospital of Jiujiang University, Jiujiang, Jiangxi 332000, P.R. China
Published online on: April 8, 2019 https://doi.org/10.3892/mmr.2019.10140
Pages: 4719-4726
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Leukemia inhibitory factor (LIF) modulates various biological processes. Although previous studies have described the effects of LIF on adipocyte differentiation, the role of LIF receptor (LIFR) on adipocyte differentiation remains unclear. Using reverse transcription‑quantitative PCR (RT‑qPCR), LIFR expression was demonstrated to increase during adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs), indicating that LIFR may be involved in this process. To further evaluate the association between LIFR and adipogenic differentiation, lentivirus‑mediated LIFR knockdown was performed in hMSCs. Cells were divided into two groups: Negative control group and LIFR‑knockdown group. During the adipogenic differentiation process, intracellular lipid accumulation was assessed with Oil Red O staining at various time points (days 3, 6 and 9). Additionally, the mRNA and protein expression levels of LIF, LIFR and three molecular indicators of adipogenesis, peroxisome proliferator‑activated receptor γ (PPARγ), CCAAT enhancer binding protein α (C/EBPα) and fatty acid binding protein 4 (FABP4/aP2), were assessed by RT‑qPCR and western blotting. The culture supernatant was collected to evaluate the concentration of LIF using ELISA. The present results suggested that LIFR expression progressively increased during adipogenic differentiation of hMSCs. Conversely, LIFR knockdown significantly suppressed this process. Additionally, PPARγ, C/EBPα and aP2 were inhibited following LIFR knockdown. In contrast with LIFR, the expression levels of LIF were significantly decreased after the initiation of adipogenic differentiation. Therefore, the expression levels of LIF and LIFR exhibited opposite trends. Collectively, the present results suggested that LIFR promoted adipogenic differentiation, whereas LIF may negatively regulate this process.

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1	Kopelman PG: Obesity as a medical problem. Nature. 404:635–643. 2000. View Article : Google Scholar : PubMed/NCBI
2	Kahn BB and Flier JS: Obesity and insulin resistance. J Clin Invest. 106:473–481. 2000. View Article : Google Scholar : PubMed/NCBI
3	Keller M, Hopp L, Liu X, Wohland T, Rohde K, Cancello R, Klös M, Bacos K, Kern M, Eichelmann F, et al: Genome-wide DNA promoter methylation and transcriptome analysis in human adipose tissue unravels novel candidate genes for obesity. Mol Metab. 6:86–100. 2016. View Article : Google Scholar : PubMed/NCBI
4	Otto TC and Lane MD: Adipose development: From stem cell to adipocyte. Crit Rev Biochem Mol Biol. 40:229–242. 2005. View Article : Google Scholar : PubMed/NCBI
5	Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S and Marshak DR: Multilineage potential of adult human mesenchymal stem cells. Science. 284:143–147. 1999. View Article : Google Scholar : PubMed/NCBI
6	Helder MN, Knippenberg M, Klein-Nulend J and Wuisman PI: Stem cells from adipose tissue allow challenging new concepts for regenerative medicine. Tissue Eng. 13:1799–1808. 2007. View Article : Google Scholar : PubMed/NCBI
7	Barry F, Boynton RE, Liu B and Murphy JM: Chondrogenic differentiation of mesenchymal stem cells from bone marrow: Differentiation-dependent gene expression of matrix components. Exp Cell Res. 268:189–200. 2001. View Article : Google Scholar : PubMed/NCBI
8	Arinzeh TL: Mesenchymal stem cells for bone repair: Preclinical studies and potential orthopedic applications. Foot Ankle Clin. 10:651–665. 2005. View Article : Google Scholar : PubMed/NCBI
9	Subash-Babu P and Alshatwi AA: Aloe-emodin inhibits adipocyte differentiation and maturation during in vitro human mesenchymal stem cell adipogenesis. J Biochem Mol Toxicol. 26:291–300. 2012. View Article : Google Scholar : PubMed/NCBI
10	del Valle I, Rudloff S, Carles A, Li Y, Liszewska E, Vogt R and Kemler R: E-cadherin is required for the proper activation of the Lifr/Gp130 signaling pathway in mouse embryonic stem cells. Development. 140:1684–1692. 2013. View Article : Google Scholar : PubMed/NCBI
11	Pan W, Cain C, Yu Y and Kastin AJ: Receptor-mediated transport of LIF across blood-spinal cord barrier is upregulated after spinal cord injury. J Neuroimmunol. 174:119–125. 2006. View Article : Google Scholar : PubMed/NCBI
12	Plun-Favreau H, Perret D, Diveu C, Froger J, Chevalier S, Lelièvre E, Gascan H and Chabbert M: Leukemia inhibitory factor (LIF), cardiotrophin-1, and oncostatin M share structural binding determinants in the immunoglobulin-like domain of LIF receptor. J Biol Chem. 278:27169–27179. 2003. View Article : Google Scholar : PubMed/NCBI
13	White UA and Stephens JM: Neuropoietin activates STAT3 Independent of LIFR activation in adipocytes. Biochem Biophys Res Commun. 395:48–50. 2010. View Article : Google Scholar : PubMed/NCBI
14	Morton SD, Cadamuro M, Brivio S, Vismara M, Stecca T, Massani M, Bassi N, Furlanetto A, Joplin RE, Floreani A, et al: Leukemia inhibitory factor protects cholangiocarcinoma cells from drug-induced apoptosis via a PI3K/AKT-dependent Mcl-1 activation. Oncotarget. 6:26052–26064. 2015. View Article : Google Scholar : PubMed/NCBI
15	Ikeda S, Itoh S, Yamamoto Y, Matsushita K, Naruse H and Hayashi M: Developmental stage-dependent effects of leukemia inhibitory factor on adipocyte differentiation of murine bone marrow stromal cells. Cell Biochem Biophys. 74:11–17. 2016. View Article : Google Scholar : PubMed/NCBI
16	Aubert J, Dessolin S, Belmonte N, Li M, McKenzie FR, Staccini L, Villageois P, Barhanin B, Vernallis A, Smith AG, et al: Leukemia inhibitory factor and its receptor promote adipocyte differentiation via the mitogen-activated protein kinase cascade. J Biol Chem. 274:24965–24972. 1999. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
18	Fernyhough ME, Hausman GJ, Guan LL, Okine E, Moore SS and Dodson MV: Mature adipocytes may be a source of stem cells for tissue engineering. Biochem Biophys Res Commun. 368:455–457. 2008. View Article : Google Scholar : PubMed/NCBI
19	Li X, Yang Y, Yan R, Xu X, Gao L, Mei J, Liu J, Wang X, Zhang J, Wu P, et al: miR-377-3p regulates adipogenic differentiation of human bone marrow mesenchymal stem cells by regulating LIFR. Mol Cell Biochem. 449:295–303. 2018. View Article : Google Scholar : PubMed/NCBI
20	Farmer SR: Transcriptional control of adipocyte formation. Cell Metab. 4:263–273. 2006. View Article : Google Scholar : PubMed/NCBI
21	Kudo M, Sugawara A, Uruno A, Takeuchi K and Ito S: Transcription suppression of peroxisome proliferator-activated receptor gamma2 gene expression by tumor necrosis factor alpha via an inhibition of CCAAT/enhancer-binding protein delta during the early stage ofadipocyte differentiation. Endocrinology. 145:4948–4956. 2004. View Article : Google Scholar : PubMed/NCBI
22	Rosen ED, Walkey CJ, Puigserver P and Spiegelman BM: Transcriptional regulation of adipogenesis. Genes Dev. 14:1293–1307. 2000.PubMed/NCBI
23	Zhang J, Huang Y, Shao H, Bi Q, Chen J and Ye Z: Grape seed procyanidin B2 inhibits adipogenesis of 3T3-L1 cells by targeting peroxisome proliferator-activated receptor γ with miR-483-5p involved mechanism. Biomed Pharmacother. 86:292–296. 2017. View Article : Google Scholar : PubMed/NCBI
24	Floyd ZE and Stephens JM: Controlling a master switch of adipocyte development and insulin sensitivity: Covalent modifications of PPARγ. Biochim Biophys Acta. 1822:1090–1095. 2012. View Article : Google Scholar : PubMed/NCBI
25	Furuhashi M, Tuncman G, Gorgun CZ, Makowski L, Atsumi G, Vaillancourt E, Kono K, Babaev VR, Fazio S, Linton MF, et al: Treatment of diabetes and atherosclerosis by inhibiting fatty-acid-binding protein FABP4. Nature. 447:959–965. 2007. View Article : Google Scholar : PubMed/NCBI
26	Hwang CS, Mandrup S, MacDougald OA, Geiman DE and Lane MD: Transcriptional activation of the mouse obese (ob) gene by CCAAT/enhancer binding protein alpha. Proc Natl Acad Sci USA. 93:873–877. 1996. View Article : Google Scholar : PubMed/NCBI
27	Oskowitz AZ, Lu J, Penfornis P, Ylostalo J, McBride J, Flemington EK, Prockop DJ and Pochampally R: Human multipotent stromal cells from bone marrow and microRNA: Regulation of differentiationand leukemia inhibitory factor expression. Proc Natl Acad Sci USA. 105:18372–18377. 2008. View Article : Google Scholar : PubMed/NCBI
28	Hogan JC and Stephens JM: Effects of leukemia inhibitory factor on 3T3-L1 adipocytes. J Endocrinol. 185:485–496. 2005. View Article : Google Scholar : PubMed/NCBI
29	Margioula-Siarkou C, Prapas Y, Petousis S, Milias S, Ravanos K, Kalogiannidis I, Mavromatidis G, Haitoglou C, Prapas N and Rousso D: LIF and LIF-R expression in the endometrium of fertile and infertile women: A prospective observational case-control study. Mol Med Rep. 13:4721–4728. 2016. View Article : Google Scholar : PubMed/NCBI
30	Li Y, Sun L, Zhao D, Ouyang J and Xiang M: Aberrant expression of leukemia inhibitory factor receptor (LIFR) and leukemia inhibitory factor (LIF) is associated with tubal pregnancy occurrence. Turk J Med Sci. 45:214–220. 2015. View Article : Google Scholar : PubMed/NCBI
31	Saito M, Asai Y, Imai K, Hiratoko S and Tanaka K: Connexin30.3 is expressed in mouse embryonic stem cells and is responsive to leukemia inhibitory factor. Sci Rep. 7:424032017. View Article : Google Scholar : PubMed/NCBI
32	Cherepkova MY, Sineva GS and Pospelov VA: Leukemia inhibitory factor (LIF) withdrawal activates mTOR signaling pathway in mouse embryonic stem cells through the MEK/ERK/TSC2 pathway. Cell Death Dis. 7:e20502016. View Article : Google Scholar : PubMed/NCBI
33	Cheng HC, Liu SW, Li W, Zhao XF, Zhao X, Cheng M, Qiu L and Ma J: Arsenic trioxide regulates adipogenic and osteogenic differentiation in bone marrow MSCs of aplastic anemia patients through BMP4 gene. Acta Biochim Biophys Sin (Shanghai). 47:673–679. 2015. View Article : Google Scholar : PubMed/NCBI
34	Lecourt S, Vanneaux V, Leblanc T, Leroux G, Ternaux B, Benbunan M, Chomienne C, Baruchel A, Marolleau JP, Gluckman E, et al: Bone marrow microenvironment in fanconi anemia: A prospective functional study in a cohort of fanconi anemia patients. Stem Cells Dev. 19:203–208. 2010. View Article : Google Scholar : PubMed/NCBI
35	Natesh K, Bhosale D, Desai A, Chandrika G, Pujari R, Jagtap J, Chugh A, Ranade D and Shastry P: Oncostatin-M differentially regulates mesenchymal and proneural signature genes in gliomas via STAT3 signaling. Neoplasia. 17:225–237. 2015. View Article : Google Scholar : PubMed/NCBI
36	Wagener EM, Aurich M, Aparicio-Siegmund S, Floss DM, Garbers C, Breusing K, Rabe B, Schwanbeck R, Grötzinger J, Rose-John S and Scheller J: The amino acid exchange R28E in ciliaryneurotrophic factor (CNTF) abrogates interleukin-6 receptor-dependent but retains CNTF receptor-dependent signaling via glycoprotein 130 (gp130)/leukemia inhibitory factor receptor (LIFR). J Biol Chem. 289:18442–184450. 2014. View Article : Google Scholar : PubMed/NCBI
37	Liu GX, Zhu JC, Chen XY, Zhu AZ, Liu CC, Lai Q and Chen ST: Inhibition of adipogenic differentiation of bone marrow mesenchymal stem cells by erythropoietin via activating ERK and P38 MAPK. Genet Mol Res. 14:6968–6977. 2015. View Article : Google Scholar : PubMed/NCBI