TCF4/β‑catenin complex is directly upstream of FGF21 in mouse stomach cancer cells

Pei,Jihua; Song,Na; Wu,Limin; Qi,Jinbo; Xia,Shenglong; Xu,Changlong; Zheng,Bo; Yang,Jun; Qiu,Yanyan; Wang,Haijun; Jiang,Yi

doi:10.3892/etm.2017.5493

TCF4/β‑catenin complex is directly upstream of FGF21 in mouse stomach cancer cells

Authors:
- Jihua Pei
- Na Song
- Limin Wu
- Jinbo Qi
- Shenglong Xia
- Changlong Xu
- Bo Zheng
- Jun Yang
- Yanyan Qiu
- Haijun Wang
- Yi Jiang
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Affiliations: Department of Gastroenterology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, P.R. China, Department of Molecular Biology and Biochemistry, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China, Department of Pathology, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China
Published online on: November 13, 2017 https://doi.org/10.3892/etm.2017.5493
Pages: 1041-1047

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Abstract

Fibroblast growth factor 21 (FGF21) as a member of the FGFs serves a key role in glucose homeostasis and protection of the liver, heart, kidney and skin from damage as well as cancer cell development. In addition, transcription of FGF21 is sensitive to diverse damages; however, the role of the transcriptional regulator of FGF21 in cancer cells remains to be elucidated. FGFs were identified to have dominant expression in cancer cells; therefore, mouse forestomach carcinoma (MFC) cells were used in the present study, which is a mouse stomach cancer cell strain for identifying the FGF21 regulators. In promoter analysis of FGF21, the putative transcription factor 4 (TCF4) binding motifs (T/AC/GAAAG) were observed within 1.5 kb of the promoter region. Further chromatin immunoprecipitation and yeast‑one hybrid assays identified that TCF4 directly bound to one of the two putative binding motifs observed. A co‑immunoprecipitation assay identified that β‑catenin interacts with TCF4 in MFC cells, and the β‑catenin/TCF4 complex bound to the promoter of FGF21. In order to examine the function of TCF4 and β‑catenin in transcriptional regulation of FGF21, TCF4 and β‑catenin was transiently expressed in MFC cells. Reverse transcription‑quantitative polymerase chain reaction results revealed that overexpression of TCF4 and β‑catenin activated FGF21 transcription. Besides, suppression of β‑catenin via a specific short interfering RNA resulted in reduction of FGF21 expression. Together these findings suggest that the β‑catenin/TCF complex directly activates FGF21 via promoter binding. The observations of the present study may help elucidate the regulatory mechanism of FGF21, which is a key pharmaceutical protein.

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1	Mohammadi M, Olsen SK and Ibrahimi OA: Structural basis for fibroblast growth factor receptor activation. CytokineGrowth Factor Rev. 16:107–137. 2005. View Article : Google Scholar
2	Feldman B, Poueymirou W, Papaioannou VE, DeChiara TM and Goldfarb M: Requirement of FGF-4 for postimplantation mouse development. Science. 267:246–249. 1995. View Article : Google Scholar : PubMed/NCBI
3	Dubrulle J and Pourquié O: fgf8 mRNA decay establishes a gradient that couples axial elongation to patterning in the vertebrate embryo. Nature. 427:419–422. 2004. View Article : Google Scholar : PubMed/NCBI
4	Sun X, Meyers EN, Lewandoski M and Martin GR: Targeted disruption of Fgf8 causes failure of cell migration in the gastrulating mouse embryo. Genes Dev. 13:1834–1846. 1999. View Article : Google Scholar : PubMed/NCBI
5	Martin GR: The roles of FGFs in the early development of vertebrate limbs. Genes Dev. 12:1571–1586. 1998. View Article : Google Scholar : PubMed/NCBI
6	Goldfarb M: Functions of fibroblast growth factors in vertebrate development. Cytokine Growth Factor Rev. 7:311–325. 1996. View Article : Google Scholar : PubMed/NCBI
7	Kanazawa S, Fujiwara T, Matsuzaki S, Shingaki K, Taniguchi M, Miyata S, Tohyama M, Sakai Y, Yano K, Hosokawa K and Kubo T: bFGF regulates PI3-kinase-Rac1-JNK pathway and promotes fibroblast migration in wound healing. PloS One. 5:e122282010. View Article : Google Scholar : PubMed/NCBI
8	Yie J, Wang W, Deng L, Tam LT, Stevens J, Chen MM, Li Y, Xu J, Lindberg R, Hecht R, et al: Understanding the physical interactions in the FGF21/FGFR/beta-Klotho complex: Structural requirements and implications in FGF21 signaling. Chem Biol Drug Des. 79:398–410. 2012. View Article : Google Scholar : PubMed/NCBI
9	Belov AA and Mohammadi M: Molecular mechanisms of fibroblast growth factor signaling in physiology and pathology. Cold Spring Harb Perspect Biol. 5:a0159582013. View Article : Google Scholar : PubMed/NCBI
10	Nishimura T, Nakatake Y, Konishi M and Itoh N: Identification of a novel FGF, FGF-21, preferentially expressed in the liver. Biochim Biophys Acta. 1492:203–206. 2000. View Article : Google Scholar : PubMed/NCBI
11	Liang Q, Zhong L, Zhang J, Wang Y, Bornstein SR, Triggle CR, Ding H, Lam KS and Xu A: FGF21 maintains glucose homeostasis by mediating the cross talk between liver and brain during prolonged fasting. Diabetes. 63:4064–4075. 2014. View Article : Google Scholar : PubMed/NCBI
12	Lin Z, Tian H, Lam KS, Lin S, Hoo RC, Konishi M, Itoh N, Wang Y, Bornstein SR, Xu A and Li X: Adiponectin mediates the metabolic effects of FGF21 on glucose homeostasis and insulin sensitivity in mice. Cell Metab. 17:779–789. 2013. View Article : Google Scholar : PubMed/NCBI
13	Lin Z, Wu F, Lin S, Pan X, Jin L, Lu T, Shi L, Wang Y, Xu A and Li X: Adiponectin protects against acetaminophen-induced mitochondrial dysfunction and acute liver injury by promoting autophagy in mice. J Hepatol. 61:825–831. 2014. View Article : Google Scholar : PubMed/NCBI
14	Song Y, Ding J, Jin R, Jung J, Li S, Yang J, Wang A and Li Z: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration. Mol Med Rep. 13:3619–3626. 2016. View Article : Google Scholar : PubMed/NCBI
15	Song YH, Zhu YT, Ding J, Zhou FY, Xue JX, Jung JH, Li ZJ and Gao WY: Distribution of fibroblast growth factors and their roles in skin fibroblast cell migration. Mol Med Rep. 14:3336–3342. 2016. View Article : Google Scholar : PubMed/NCBI
16	Gong Q, Hu Z, Zhang F, Cui A, Chen X, Jiang H, Gao J, Chen X, Han Y, Liang Q, et al: Fibroblast growth factor 21 improves hepatic insulin sensitivity by inhibiting mammalian target of rapamycin complex 1 in mice. Hepatology. 64:425–438. 2016. View Article : Google Scholar : PubMed/NCBI
17	Inagaki T, Dutchak P, Zhao G, Ding X, Gautron L, Parameswara V, Li Y, Goetz R, Mohammadi M, Esser V, et al: Endocrine regulation of the fasting response by PPARalpha-mediated induction of fibroblast growth factor 21. Cell Metab. 5:415–425. 2007. View Article : Google Scholar : PubMed/NCBI
18	Wang Y, Solt LA and Burris TP: Regulation of FGF21 expression and secretion by retinoic acid receptor-related orphan receptor alpha. J Biol Chem. 285:15668–15673. 2010. View Article : Google Scholar : PubMed/NCBI
19	Li Y, Wong K, Walsh K, Gao B and Zang M: Retinoic acid receptor β stimulates hepatic induction of fibroblast growth factor 21 to promote fatty acid oxidation and control whole-body energy homeostasis in mice. J Biol Chem. 288:10490–10504. 2013. View Article : Google Scholar : PubMed/NCBI
20	Shtutman M, Zhurinsky J, Simcha I, Albanese C, D'Amico M, Pestell R and Ben-Ze'ev A: The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci USA. 96:pp. 5522–5527. 1999; View Article : Google Scholar : PubMed/NCBI
21	Moon RT, Kohn AD, De Ferrari GV and Kaykas A: WNT and β-catenin signalling: Diseases and therapies. Nat Rev Genet. 5:691–701. 2004. View Article : Google Scholar : PubMed/NCBI
22	Clevers H: Wnt/beta-catenin signaling in development and disease. Cell. 127:469–480. 2006. View Article : Google Scholar : PubMed/NCBI
23	Brack AS, Conboy MJ, Roy S, Lee M, Kuo CJ, Keller C and Rando TA: Increased Wnt signaling during aging alters muscle stem cell fate and increases fibrosis. Science. 317:807–810. 2007. View Article : Google Scholar : PubMed/NCBI
24	Gordon MD and Nusse R: Wnt signaling: Multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem. 281:22429–22433. 2006. View Article : Google Scholar : PubMed/NCBI
25	Aberle H, Bauer A, Stappert J, Kispert A and Kemler R: beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J. 16:3797–3804. 1997. View Article : Google Scholar : PubMed/NCBI
26	Schuijers J, Mokry M, Hatzis P, Cuppen E and Clevers H: Wnt-induced transcriptional activation is exclusively mediated by TCF/LEF. EMBO J. 33:146–156. 2014. View Article : Google Scholar : PubMed/NCBI
27	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-tie quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
28	Dutchak PA, Katafuchi T, Bookout AL, Choi JH, Yu RT, Mangelsdorf DJ and Kliewer SA: Fibroblast growth factor-21 regulates PPARγ activity and the antidiabetic actions of thiazolidinediones. Cell. 148:556–567. 2012. View Article : Google Scholar : PubMed/NCBI
29	Fisher FM, Kleiner S, Douris N, Fox EC, Mepani RJ, Verdeguer F, Wu J, Kharitonenkov A, Flier JS, Maratos-Flier E and Spiegelman BM: FGF21 regulates PGC-1α and browning of white adipose tissues in adaptive thermogenesis. Genes Dev. 26:271–281. 2012. View Article : Google Scholar : PubMed/NCBI
30	Shu W, Guttentag S, Wang Z, Andl T, Ballard P, Lu MM, Piccolo S, Birchmeier W, Whitsett JA, Millar SE and Morrisey EE: Wnt/beta-catenin signaling acts upstream of N-myc, BMP4, and FGF signaling to regulate proximal-distal patterning in the lung. Dev Biol. 283:226–239. 2005. View Article : Google Scholar : PubMed/NCBI