Genistein induces adipogenic differentiation in human bone marrow mesenchymal stem cells and suppresses their osteogenic potential by upregulating PPARγ

Zhang,Li‑Yan; Xue,Hao‑Gang; Chen,Ji‑Ying; Chai,Wei; Ni,Ming

doi:10.3892/etm.2016.3120

Genistein induces adipogenic differentiation in human bone marrow mesenchymal stem cells and suppresses their osteogenic potential by upregulating PPARγ

Authors:
- Li‑Yan Zhang
- Hao‑Gang Xue
- Ji‑Ying Chen
- Wei Chai
- Ming Ni
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Affiliations: Department of Orthopedics, General Hospital of Chinese People's Liberation Army, Beijing 100853, P.R. China, First Department of Orthopedics, The Affiliated Hospital of Beihua University, Jilin, Jilin 132001, P.R. China
Published online on: March 2, 2016 https://doi.org/10.3892/etm.2016.3120
Pages: 1853-1858

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Abstract

Genistein is a soy isoflavone that exists in the form of an aglycone. It is the primary active component in soy isoflavone and has a number of biological activities (anti‑inflammatory and anti‑oxidative). However, the specific effect of genistein on human bone marrow mesenchymal stem cells (BMSCs) remains unclear. In the present study, the mechanism underlying the effect of genistein on the suppression of BMSC adipogenic differentiation and the enhancement of osteogenic potential was investigated using an MTT assay. It was observed that genistein significantly increased BMSC cell proliferation in a time‑ and dose‑dependent manner (P<0.01). In addition, reverse transcription‑quantitative polymerase chain reaction revealed that genistein significantly inhibited the expression of runt‑related transcription factor 2 (Runx2), type I collagen (Col I) and osteocalcin (OC; P<0.01). Furthermore, 20 µm genistein significantly inhibited the activity of alkaline phosphatase (ALP) and increased the activity of triglycerides (TGs) increased (P<0.01) as determined by an enzyme-linked immunosorbent assay. Finally, western blotting revealed that BMSC pretreatment with 20 µm genistein significantly increased peroxisome proliferator-activated receptor γ (PPARγ) protein expression (P<0.01). This suggests that the downregulation of PPARγ may significantly reduce the effect of genistein on cell proliferation, suppress the expression of Runx2, Col I and OC mRNA, and reduce ALP and promote TG activity in BMSCs. Thus, the results of the present study conclude that genistein induces adipogenic differentiation in human BMSCs and suppresses their osteogenic potential by upregulating the expression of PPARγ. In conclusion, genistein may be a promising candidate drug for treatment against osteogenesis.

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1	Katsuda T, Tsuchiya R, Kosaka N, Yoshioka Y, Takagaki K, Oki K, Takeshita F, Sakai Y, Kuroda M and Ochiya T: Human adipose tissue-derived mesenchymal stem cells secrete functional neprilysin-bound exosomes. Sci Rep. 3:11972013. View Article : Google Scholar : PubMed/NCBI
2	Dumont N, Boyer L, Émond H, Celebi-Saltik B, Pasha R, Bazin R, Mantovani D, Roy DC and Pineault N: Medium conditioned with mesenchymal stromal cell-derived osteoblasts improves the expansion and engraftment properties of cord blood progenitors. Exp Hematol. 42:741–752.e1. 2014. View Article : Google Scholar : PubMed/NCBI
3	Kawai M, Green CB, Lecka-Czernik B, Douris N, Gilbert MR, Kojima S, Ackert-Bicknell C, Garg N, Horowitz MC, Adamo ML, et al: A circadian-regulated gene, Nocturnin, promotes adipogenesis by stimulating PPAR-gamma nuclear translocation. Proc Natl Acad Sci USA. 107:10508–10513. 2010. View Article : Google Scholar : PubMed/NCBI
4	Lv FH, Gao JZ, Teng QL and Zhang JY: Effect of folic acid and vitamin B12 on the expression of PPARγ, caspase-3 and caspase-8 mRNA in the abdominal aortas of rats with hyperlipidemia. Exp Ther Med. 6:184–188. 2013.PubMed/NCBI
5	Zhou Y, Zhu ZL, Guan XX, Hou WW and Yu HY: Reciprocal roles between caffeine and estrogen on bone via differently regulating cAMP-PKA pathway: The possible mechanism for caffeine-induced osteoporosis in women and estrogens antagonistic effects. Med Hypotheses. 73:83–85. 2009. View Article : Google Scholar : PubMed/NCBI
6	Lu T, Huang Y, Wang H, Ma Y and Guan W: Multi-lineage potential research of bone marrow-derived stromal cells (BMSCs) from cattle. Appl Biochem Biotechnol. 172:21–35. 2014. View Article : Google Scholar : PubMed/NCBI
7	Fan J, Li J and Fan Q: Naringin promotes differentiation of bone marrow stem cells into osteoblasts by upregulating the expression levels of microRNA-20a and downregulating the expression levels of PPARgamma. Mol Med Rep. 12:4759–4765. 2015.PubMed/NCBI
8	Weivoda MM and Hohl RJ: Geranylgeranyl pyrophosphate stimulates PPARγ expression and adipogenesis through the inhibition of osteoblast differentiation. Bone. 50:467–476. 2012. View Article : Google Scholar : PubMed/NCBI
9	Cao J, Ou G, Yang N, Ding K, Kream BE, Hamrick MW, Isales CM and Shi XM: Impact of targeted PPARγ disruption on bone remodeling. Mol Cell Endocrinol. 410:27–34. 2015. View Article : Google Scholar : PubMed/NCBI
10	Chen Y, Chen L, Yin Q, Gao H, Dong P, Zhang X and Kang J: Reciprocal interferences of TNF-α and Wnt1-β-catenin signaling axes shift bone marrow-derived stem cells towards osteoblast lineage after ethanol exposure. Cell Physiol Biochem. 32:755–765. 2013. View Article : Google Scholar : PubMed/NCBI
11	Hirayama K, Matsuzuka Y, Kamiya T, Ikeguchi M, Takagaki K and Itoh K: Metabolism of isoflavones found in the Pueraria thomsonii flower by human intestinal microbiota. Biosci Microflora. 30:135–140. 2011. View Article : Google Scholar : PubMed/NCBI
12	Kim SH, Kim CW, Jeon SY, Go RE, Hwang KA and Choi KC: Chemopreventive and chemotherapeutic effects of genistein, a soy isoflavone, upon cancer development and progression in preclinical animal models. Lab Anim Res. 30:143–150. 2014. View Article : Google Scholar : PubMed/NCBI
13	Nagaraju GP, Zafar SF and El-Rayes BF: Pleiotropic effects of genistein in metabolic, inflammatory, and malignant diseases. Nutr Rev. 71:562–572. 2013. View Article : Google Scholar : PubMed/NCBI
14	Menze ET, Esmat A, Tadros MG, Abdel-Naim AB and Khalifa AE: Genistein improves 3-NPA-induced memory impairment in ovariectomized rats: Impact of its antioxidant, anti-inflammatory and acetylcholinesterase modulatory properties. PLoS One. 10:e01172232015. View Article : Google Scholar : PubMed/NCBI
15	Chen J, Duan Y, Zhang X, Ye Y, Ge B and Chen J: Genistein induces apoptosis by the inactivation of the IGF-1R-p-Akt signaling pathway in MCF-7 human breast cancer cells. Food Funct. 6:995–1000. 2015. View Article : Google Scholar : PubMed/NCBI
16	Liao MH, Tai YT, Cherng YG, Liu SH, Chang YA, Lin PI and Chen RM: Genistein induces oestrogen receptor-α gene expression in osteoblasts through the activation of mitogen-activated protein kinases-NF-κB-activator protein-1 and promotes cell mineralisation. Br J Nutr. 111:55–63. 2014. View Article : Google Scholar : PubMed/NCBI
17	Zeng X, Yu SP, Taylor T, Ogle M and Wei L: Protective effect of apelin on cultured rat bone marrow mesenchymal stem cells against apoptosis. Stem Cell Res (Amst). 8:357–367. 2012. View Article : Google Scholar
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
19	Pauksch L, Hartmann S, Szalay G, Alt V and Lips KS: In vitro assessment of nanosilver-functionalized PMMA bone cement on primary human mesenchymal stem cells and osteoblasts. PLoS One. 9:e1147402014. View Article : Google Scholar : PubMed/NCBI
20	Lee S, Cho HY, Bui HT and Kang D: The osteogenic or adipogenic lineage commitment of human mesenchymal stem cells is determined by protein kinase C delta. BMC Cell Biol. 15:422014. View Article : Google Scholar : PubMed/NCBI
21	Antoniou J, Wang HT, Alaseem AM, Haglund L, Roughley PJ and Mwale F: The effect of Link N on differentiation of human bone marrow-derived mesenchymal stem cells. Arthritis Res Ther. 14:R2672012. View Article : Google Scholar : PubMed/NCBI
22	Nichols RA Jr, Niagro FD, Borke JL and Cuenin MF: Mechanical stretching of mouse calvarial osteoblasts in vitro models changes in MMP-2 and MMP-9 expression at the bone-implant interface. J Oral Implantol. May 11–2015.(Epub ahead of print). View Article : Google Scholar : PubMed/NCBI
23	Hu HM, Yang L, Wang Z, Liu YW, Fan JZ, Fan J, Liu J and Luo ZJ: Overexpression of integrin α2 promotes osteogenic differentiation of hBMSCs from senile osteoporosis through the ERK pathway. Int J Clin Exp Pathol. 6:841–852. 2013.PubMed/NCBI
24	Relic B, Zeddou M, Desoroux A, Beguin Y, de Seny D and Malaise MG: Genistein induces adipogenesis but inhibits leptin induction in human synovial fibroblasts. Lab Invest. 89:811–822. 2009. View Article : Google Scholar : PubMed/NCBI
25	Arrigoni C, De Luca P, Gilardi M, Previdi S, Broggini M and Moretti M: Direct but not indirect co-culture with osteogenically differentiated human bone marrow stromal cells increases RANKL-OPG ratio in human breast cancer cells generating bone metastases. Mol Cancer. 13:2382014. View Article : Google Scholar : PubMed/NCBI
26	Choudhary S, Goetjen A, Estus T, Jacome-Galarza CE, Aguila HL, Lorenzo J and Pilbeam C: Serum amyloid A3 secreted by preosteoclasts inhibits parathyroid hormone-stimulated cAMP signaling in murine osteoblasts. J Biol Chem. 23–Dec;2015.(Epub ahead of print). pii jbc. M115.686576. PubMed/NCBI
27	Song M, Tian X, Lu M, Zhang X, Ma K, Lv Z, Wang Z, Hu Y, Xun C, Zhang Z and Wang S: Genistein exerts growth inhibition on human osteosarcoma MG-63 cells via PPARγ pathway. Int J Oncol. 46:1131–1140. 2015.PubMed/NCBI
28	Chatterjee G, Roy D, Khemka VK, Chattopadhyay M and Chakrabarti S: Genistein, the isoflavone in soybean, causes amyloid beta peptide accumulation in human neuroblastoma cell line: Implications in Alzheimers disease. Aging Dis. 6:456–465. 2015. View Article : Google Scholar : PubMed/NCBI
29	Valles SL, Dolz-Gaiton P, Gambini J, Borras C, Lloret A, Pallardo FV and Viña J: Estradiol or genistein prevent Alzheimers disease-associated inflammation correlating with an increase PPAR gamma expression in cultured astrocytes. Brain Res. 1312:138–144. 2010. View Article : Google Scholar : PubMed/NCBI
30	Zhuang H, Zhang X, Zhu C, Tang X, Yu F, Shang GW and Cai X: Molecular mechanisms of PPARγ governing MSC osteogenic and adipogenic differentiation. Curr Stem Cell Res Ther. 31–May;2015.(Epub ahead of print).
31	Zhou H, Yang X, Wang N, Zhang Y and Cai G: Tigogenin inhibits adipocytic differentiation and induces osteoblastic differentiation in mouse bone marrow stromal cells. Mol Cell Endocrinol. 270:17–22. 2007. View Article : Google Scholar : PubMed/NCBI
32	Wang L, Li L, Gao H and Li Y: Effect of pioglitazone on transdifferentiation of preosteoblasts from rat bone mesenchymal stem cells into adipocytes. J Huazhong Univ Sci Technolog Med Sci. 32:530–533. 2012. View Article : Google Scholar : PubMed/NCBI
33	Lee NJ, Doyle KL, Sainsbury A, Enriquez RF, Hort YJ, Riepler SJ, Baldock PA and Herzog H: Critical role for Y1 receptors in mesenchymal progenitor cell differentiation and osteoblast activity. J Bone Miner Res. 25:1736–1747. 2010. View Article : Google Scholar : PubMed/NCBI