Genistein and menaquinone-4 treatment-induced alterations in the expression of mRNAs and their products are beneficial to osteoblastic MC3T3-E1 cell functions

Katsuyama,Midori; Demura,Masashi; Katsuyama,Hironobu; Tanii,Hideji; Saijoh,Kiyofumi

doi:10.3892/mmr.2017.6632

Genistein and menaquinone-4 treatment-induced alterations in the expression of mRNAs and their products are beneficial to osteoblastic MC3T3-E1 cell functions

Authors:
- Midori Katsuyama
- Masashi Demura
- Hironobu Katsuyama
- Hideji Tanii
- Kiyofumi Saijoh
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Affiliations: Department of Hygiene, Kanazawa University School of Medicine, Kanazawa, Ishikawa 920‑8640, Japan, Department of Public Health, Kawasaki Medical University, Kurashiki, Okayama 701‑0192, Japan
Published online on: May 25, 2017 https://doi.org/10.3892/mmr.2017.6632
Pages: 873-880

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Abstract

The aim of the present study was to determine the molecular basis of the beneficial effects of genistein and/or menaquinone‑4 (MK‑4) on bone quality. Initially, 1 µM genistein was applied to MC3T3‑E1 cells for 24 h and the upregulated mRNAs that were detected by microarray were selected for further examination by reverse transcription‑quantitative‑polymerase chain reaction. Among them, alterations were observed in the level of GATA‑binding protein 6 (GATA6), Notch gene homolog 2 (NOTCH2), Wnt family member 5A (WNT5A), bone γ‑carboxyglutamate protein (BGLAP), chondroadherin (CHAD), dipeptidyl peptidase 4 (DPP4), ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), alkaline phosphatase (ALP) 3 and ATPase phospholipid‑transporting 11A (ATP11A) in response to treatment with 0.1 µM 17‑β‑estradiol, 1 µM genistein, and/or 1 µM MK‑4. GATA6, NOTCH2 and WNT5A are considered to be associated with osteoclast, but not osteoblast, function; however, increases in osteoblastic mRNAs, including BGLAP and CHAD, were observed in each of the treatment groups at 48 h. Immunocytochemical analysis confirmed an increase in CHAD and DPP4 proteins following the administration of genistein + MK‑4. Furthermore, genistein + MK‑4 led to alterations in cell morphology to spindle or oval shapes, and increased the intensity of ALP staining. Although the level of ALP mRNA was not consistently altered in response to the treatments, a marked increase in ALP activity was observed following 96 h treatment with genistein + MK‑4. Therefore, the simultaneous intake of genistein and MK‑4 appears to be beneficial for the maintenance of bone quality.

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1	Milgram JW: Adult bone structure. Radiologic and Histrogic Pathology of Nontumorous Diseases of Bone and Joints. 1. Illinoi, USA: Northbrook Publishing Co. Inc.; pp. 1–5. 1990
2	Czernick B: Morphology of normal bone. Dolfman and Czernick's Bone Tumors. 2nd. Philadelphia, USA: Elsevier; pp. 3–12. 1998
3	Matsuo K and Irie N: Osteoclast-osteoblast communication. Arch Biochem Biophys. 473:201–209. 2008. View Article : Google Scholar : PubMed/NCBI
4	Turner RT, Riggs BL and Spelsberg TC: Skeletal effects of estrogen. Endocr Rev. 15:275–300. 1994. View Article : Google Scholar : PubMed/NCBI
5	Ballane G, Cauley JA, Luckey MM and Fuleihan Gel-H: Secular trends in hip fractures worldwide: Opposing trends East versus West. J Bone Miner Res. 29:1745–1755. 2014. View Article : Google Scholar : PubMed/NCBI
6	Zaheer K and Akhtar Humayoun MH: An updated review of dietary isoflavones: Nutrition, processing, bioavailability and impacts on human health. Crit Rev Food Sci Nutr. 57:1280–1293. 2017. View Article : Google Scholar : PubMed/NCBI
7	Hertrampf T, Gruca MJ, Seibel J, Laudenbach U, Fritzemeier KH and Diel P: The bone-protective effect of the phytoestrogen genistein is mediated via ER alpha-dependent mechanisms and strongly enhanced by physical activity. Bone. 40:1529–1535. 2007. View Article : Google Scholar : PubMed/NCBI
8	Hur HG, Lay JO Jr, Beger RD, Freeman JP and Rafii F: Isolation of human intestinal bacteria metabolizing the natural isoflavone glycosides daidzin and genistin. Arch Microbiol. 174:422–428. 2000. View Article : Google Scholar : PubMed/NCBI
9	Katsuyama H, Arii M, Hinenoya H, Matsushima M, Fushimi S, Tomita M, Okuyama T, Hidaka K, Watanabe Y, Tamechika Y and Saijoh K: Alterations in bone turnover by isoflavone aglycone supplementation in relation to estrogen receptor α polymorphism. Mol Med Rep. 3:531–535. 2010. View Article : Google Scholar : PubMed/NCBI
10	Kaneki M, Hodges SJ, Hosoi T, Fujiwara S, Lyons A, Crean SJ, Ishida N, Nakagawa M, Takechi M, Sano Y, et al: Japanese fermented soybean food as the major determinant of the large geographic difference in circulating levels of Vitamin K2: Possible implications for hip-fracture risk. Nutrition. 17:315–321. 2001. View Article : Google Scholar : PubMed/NCBI
11	Katsuyama H, Ideguchi S, Fukunaga M, Fukunaga T, Saijoh K and Sunami S: Promotion of bone formation by fermented soybean (Natto) intake in premenopausal women. J Nutr Sci Vitaminol (Tokyo). 50:114–120. 2004. View Article : Google Scholar : PubMed/NCBI
12	Tsangalis D, Wilcox G, Shah NP and Stojanovska L: Bioavailability of isoflavone phytoestrogens in postmenopausal women consuming soya milk fermented with probiotic bifidobacteria. Br J Nutr. 93:867–877. 2005. View Article : Google Scholar : PubMed/NCBI
13	Schurgers LJ and Vermeer C: Determination of phylloquinone and menaquinones in food. Effect of food matrix on circulating vitamin K concentrations. Haemostasis. 30:298–307. 2000.PubMed/NCBI
14	Tsukamoto Y, Ichise H, Kakuda H and Yamaguchi M: Intake of fermented soybean (natto) increases circulating vitamin K2 (menaquinone-7) and gamma-carboxylated osteocalcin concentration in normal individuals. J Bone Miner Metab. 18:216–222. 2000. View Article : Google Scholar : PubMed/NCBI
15	Elder SJ, Haytowitz DB, Howe J, Peterson JW and Booth SL: Vitamin K contents of meat, dairy, and fast food in the U.S. Diet. J Agric Food Chem. 54:463–467. 2006. View Article : Google Scholar : PubMed/NCBI
16	Sato T, Schurgers LJ and Uenishi K: Comparison of menaquinone-4 and menaquinone-7 bioavailability in healthy women. Nutr J. 11:932012. View Article : Google Scholar : PubMed/NCBI
17	Davis L, Kuehl M and Battey J: Selection of Poly-A⁺ RNA on Oligo-dT Cellulose. Basic Methods in Molecular Biology. 2nd. Connecticut, USA: Appleton & Lange; pp. 344–349. 1994
18	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
19	Dawson-Saunders B and Trapp RG: Comparing three or more means. IN basic and clinical biostatistics Connecticut, USA: Appleton & Lange; pp. 124–141. 1990
20	Kuiper GG, Lemmen JG, Carlsson B, Corton JC, Safe SH, van der Saag PT, van der Burg B and Gustafsson JK: Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor β. Endocrinology. 139:4252–4263. 1998. View Article : Google Scholar : PubMed/NCBI
21	Takahashi T, Saito A, Hashimoto S and Sato C: Isoflavone contents of soybean and soy-based foods produced in Hokkaido. Rep Hokkaido Inst Pub Hlth. 52:29–36. 2002.
22	Standard tables of food composition in Japan, . 7th. Ministry of Education Culture, Sports, Science and Technology. Japan: 2016, (In Japanese).
23	Bhushan R, Grünhagen J, Becker J, Robinson PN, Ott CE and Knaus P: miR-181a promotes osteoblastic differentiation through repression of TGF-β signaling molecules. Int J Biochem Cell Biol. 45:696–705. 2013. View Article : Google Scholar : PubMed/NCBI
24	Zanotti S and Canalis E: Notch signaling and the skeleton. Endocr Rev. 37:223–253. 2016. View Article : Google Scholar : PubMed/NCBI
25	Keller KC, Ding H, Tieu R, Sparks NR, Ehnes DD and Nieden Zur NI: Wnt5a supports osteogenic lineage decisions in embryonic stem cells. Stem Cells Dev. 25:1020–1032. 2016. View Article : Google Scholar : PubMed/NCBI
26	Kobayashi Y, Uehara S, Udagawa N and Takahashi N: Regulation of bone metabolism by Wnt signals. J Biochem. 159:387–392. 2016. View Article : Google Scholar : PubMed/NCBI
27	Yamaguchi M: Nutritional factors and bone homeostasis: Synergistic effect with zinc and genistein in osteogenesis. Mol Cell Biochem. 366:201–221. 2012. View Article : Google Scholar : PubMed/NCBI
28	Katsuyama H, Fushimi S, Yamane K, Watanabe Y, Shimoya K, Okuyama T, Katsuyama M, Saijoh K and Tomita M: Effect of vitamin K2 on the development of stress-induced osteopenia in a growing senescence-accelerated mouse prone 6 strain. Exp Ther Med. 10:843–850. 2015.PubMed/NCBI
29	Shikano K, Kaneko K, Kawazoe M, Kaburaki M, Hasunuma T and Kawai S: Efficacy of vitamin K2 for glucocorticoid-induced osteoporosis in patients with systemic autoimmune diseases. Intern Med. 55:1997–2003. 2016. View Article : Google Scholar : PubMed/NCBI
30	Capulli M, Olstad OK, Onnerfjord P, Tillgren V, Muraca M, Gautvik KM, Heinegård D, Rucci N and Teti A: The C-terminal domain of chondroadherin: A new regulator of osteoclast motility counteracting bone loss. J Bone Miner Res. 29:1833–1846. 2014. View Article : Google Scholar : PubMed/NCBI
31	Meier C, Schwartz AV, Egger A and Lecka-Czernik B: Effects of diabetes drugs on the skeleton. Bone. 82:93–100. 2016. View Article : Google Scholar : PubMed/NCBI
32	Hausmann J, Kamtekar S, Christodoulou E, Day JE, Wu T, Fulkerson Z, Albers HM, van Meeteren LA, Houben AJ, van Zeijl L, et al: Structural basis for substrate discrimination and integrin binding by autotaxin. Nat Struct Mol Biol. 18:198–204. 2011. View Article : Google Scholar : PubMed/NCBI
33	Orriss IR, Key ML, Hajjawi MO, Millán JL and Arnett TR: Acidosis is a key regulator of osteoblast ecto-nucleotidase pyrophosphatase/phosphodiesterase 1 (NPP1) expression and activity. J Cell Physiol. 230:3049–3056. 2015. View Article : Google Scholar : PubMed/NCBI
34	ATP11A ATPase phospholipid transporting 11A. Gene ID: 23250. https://www.ncbi.nlm.nih.gov/gene/23250