The BHLH transcription factor DEC1 plays an important role in the epithelial-mesenchymal transition of pancreatic cancer

Wu,Yunyan; Sato,Fuyuki; Yamada,Toshiyuki; Bhawal,Ujjal  Kumar; Kawamoto,Takeshi; Fujimoto,Katsumi; Noshiro,Mitsuhide; Seino,Hiroko; Morohashi,Satoko; Hakamada,Kenichi; Abiko,Yoshimitsu; Kato,Yukio; Kijima,Hiroshi

doi:10.3892/ijo.2012.1559

The BHLH transcription factor DEC1 plays an important role in the epithelial-mesenchymal transition of pancreatic cancer

Authors:
- Yunyan Wu
- Fuyuki Sato
- Toshiyuki Yamada
- Ujjal Kumar Bhawal
- Takeshi Kawamoto
- Katsumi Fujimoto
- Mitsuhide Noshiro
- Hiroko Seino
- Satoko Morohashi
- Kenichi Hakamada
- Yoshimitsu Abiko
- Yukio Kato
- Hiroshi Kijima
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Affiliations: Department of Pathology and Bioscience, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan, Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan, Department of Biochemistry and Molecular Biology Nihon University School of Dentistry at Matsudo, Chiba 271-8587, Japan, Department of Dental and Medical Biochemistry, Hiroshima University Graduate School of Biomedical Science, Hiroshima 734-8553, Japan, Department of Gastrointestinal Surgery, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan
Published online on: July 18, 2012 https://doi.org/10.3892/ijo.2012.1559
Pages: 1337-1346

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Abstract

DEC1 (BHLHE40/Stra13/Sharp2) is a basic helix-loop-helix (bHLH) transcription factor that is involved in the regulation of apoptosis and cell proliferation and the response to hypoxia. Epithelial-mesenchymal transition (EMT) is an important step leading to invasion and migration of various tumor cells, and TGF-β treatment has been shown to induce cancer cells to undergo EMT. However, the significance of DEC1 in TGF-β-induced EMT remains unknown. We examined the role of DEC1 in EMT of PANC-1 cells, a human pancreatic cancer cell line. As a result, we found that DEC1 was upregulated by TGF-β in PANC-1 cells, and regulated the expression and the levels of nuclear, cytoplasmic or membrane localization of EMT-related factors, including phosphorylated Smad3 (pSmad3), snail, claudin-4 and N-cadherin. In the presence of TGF-β, DEC1 knockdown by siRNA inhibited morphological changes during EMT processes, while TGF-β induced PANC-1 cells to taken on a spindle-shaped morphology. Furthermore, a combination treatment of DEC1 expression with TGF-β was closely linked to the migration and invasion of PANC-1 cells. Immunohistochemically, DEC1 and pSmad3 were detected within pancreatic cancer tissues, whereas claudin-4 expression was weaker in the cancer tissues compared with the adjacent non-cancer pancreatic tissues. These findings suggest that DEC1 plays an important role in the regulation of these EMT-related factors in pancreatic cancer.

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1	Thiery JP and Sleeman JP: Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol. 7:131–142. 2006. View Article : Google Scholar : PubMed/NCBI
2	Zavadil J and Böttinger EP: TGF-beta and epithelial-to-mesenchymal transitions. Oncogene. 24:5764–5774. 2005. View Article : Google Scholar : PubMed/NCBI
3	Miettinen PJ, Ebner R, Lopez AR and Derynck R: TGF-beta induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors. J Cell Biol. 127:2021–2036. 1994. View Article : Google Scholar : PubMed/NCBI
4	Moustakas A and Heldin CH: Signaling networks guiding epithelial-mesenchymal transitions during embryogenesis and cancer progression. Cancer Sci. 98:1512–1520. 2007. View Article : Google Scholar : PubMed/NCBI
5	Miyazawa K, Shinozaki M, Hara T, et al: Two major Smad pathways in TGF-beta superfamily signalling. Genes Cells. 7:1191–1204. 2002. View Article : Google Scholar : PubMed/NCBI
6	Giehl K, Seidel B, Gierschik P, et al: TGFbeta1 represses proliferation of pancreatic carcinoma cells which correlates with Smad4-independent inhibition of ERK activation. Oncogene. 19:4531–4541. 2000. View Article : Google Scholar : PubMed/NCBI
7	Hahn SA, Schutte M, Hoque AT, et al: DPC4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science. 271:350–353. 1996. View Article : Google Scholar : PubMed/NCBI
8	Bartsch D, Hahn SA, Danichevski KD, et al: Mutations of the DPC4/Smad4 gene in neuroendocrine pancreatic tumors. Oncogene. 18:2367–2371. 1999. View Article : Google Scholar : PubMed/NCBI
9	Massagué J: How cells read TGF-beta signals. Nat Rev Mol Cell Biol. 1:169–178. 2000.PubMed/NCBI
10	Zawel L, Dai JL, Buckhaults P, et al: Human Smad3 and Smad4 are sequence-specific transcription activators. Mol Cell. 1:611–617. 1998. View Article : Google Scholar : PubMed/NCBI
11	Ohashi S, Natsuizaka M, Naganuma S, et al: A NOTCH3-mediated squamous cell differentiation program limits expansion of EMT-competent cells that express the ZEB transcription factors. Cancer Res. 71:6836–6847. 2011. View Article : Google Scholar : PubMed/NCBI
12	Dave N, Guaita-Esteruelas S, Gutarra S, et al: Functional cooperation between Snail1 and twist in the regulation of ZEB1 expression during epithelial to mesenchymal transition. J Biol Chem. 286:12024–12032. 2011. View Article : Google Scholar : PubMed/NCBI
13	Gregory PA, Bracken CP, Smith E, et al: An autocrine TGF-beta/ ZEB/miR-200 signaling network regulates establishment and maintenance of epithelial-mesenchymal transition. Mol Biol Cell. 22:1686–1698. 2011. View Article : Google Scholar : PubMed/NCBI
14	Hong J, Zhou J, Fu J, et al: Phosphorylation of serine 68 of Twist1 by MAPKs stabilizes Twist1 protein and promotes breast cancer cell invasiveness. Cancer Res. 71:3980–3990. 2011. View Article : Google Scholar : PubMed/NCBI
15	Ikenouchi J, Matsuda M, Furuse M and Tsukita S: Regulation of tight junctions during the epithelium-mesenchyme transition: direct repression of the gene expression of claudins/occludin by Snail. J Cell Sci. 116:1959–1967. 2003. View Article : Google Scholar
16	Ohkubo T and Ozawa M: The transcription factor Snail downregulates the tight junction components independently of E-cadherin downregulation. J Cell Sci. 117:1675–1685. 2004. View Article : Google Scholar : PubMed/NCBI
17	Zavadil J, Cermak L, Soto-Nieves N, et al: Integration of TGF-beta/Smad and Jagged1/Notch signalling in epithelial-to-mesenchymal transition. EMBO J. 23:1155–1165. 2004. View Article : Google Scholar : PubMed/NCBI
18	Wu Y, Sato F, Bhawal UK, et al: Basic helix-loop-helix transcription factors DEC1 and DEC2 regulate the paclitaxel-induced apoptotic pathway of MCF-7 human breast cancer cells. Int J Mol Med. 27:491–495. 2011.PubMed/NCBI
19	Liu Y, Sato F, Kawamoto T, et al: Anti-apoptotic effect of the basic helix-loop-helix (bHLH) transcription factor DEC2 in human breast cancer cells. Genes Cells. 15:315–325. 2010. View Article : Google Scholar : PubMed/NCBI
20	Bhawal UK, Sato F, Arakawa Y, et al: Basic helix-loop-helix transcription factor DEC1 negatively regulates cyclin D1. J Pathol. 224:420–429. 2011. View Article : Google Scholar : PubMed/NCBI
21	Li Y, Xie M, Yang J, et al: The expression of antiapoptotic protein survivin is transcriptionally upregulated by DEC1 primarily through multiple sp1 binding sites in the proximal promoter. Oncogene. 25:3296–3306. 2006. View Article : Google Scholar : PubMed/NCBI
22	Turley H, Wykoff CC, Troup S, et al: The hypoxia-regulated transcription factor DEC1 (Stra13, SHARP-2) and its expression in human tissues and tumours. J Pathol. 203:808–813. 2004. View Article : Google Scholar : PubMed/NCBI
23	Chakrabarti J, Turley H, Campo L, et al: The transcription factor DEC1 (stra13, SHARP2) is associated with the hypoxic response and high tumour grade in human breast cancers. Br J Cancer. 91:954–958. 2004. View Article : Google Scholar : PubMed/NCBI
24	Giatromanolaki A, Koukourakis MI, Sivridis E, et al: DEC1 (STRA13) protein expression relates to hypoxia- inducible factor 1-alpha and carbonic anhydrase-9 overexpression in non-small cell lung cancer. J Pathol. 200:222–228. 2003. View Article : Google Scholar : PubMed/NCBI
25	Kondo J, Sato F, Kusumi T, et al: Claudin-1 expression is induced by tumor necrosis factor-α in human pancreatic cancer cells. Int J Mol Med. 22:645–649. 2008.
26	Suzuki T, Sato F, Kondo J, et al: Period is involved in the proliferation of human pancreatic MIA-PaCa2 cancer cells by TNF-alpha. Biomed Res. 29:99–103. 2008. View Article : Google Scholar : PubMed/NCBI
27	Sato F, Wu Y, Bhawal UK, Liu Y, et al: PERIOD1 (PER1) has anti-apoptotic effects, and PER3 has pro-apoptotic effects during cisplatin (CDDP) treatment in human gingival cancer CA9-22 cells. Eur J Cancer. 47:1747–1758. 2011. View Article : Google Scholar : PubMed/NCBI
28	Li Y, Zhang H, Xie M, et al: Abundant expression of Dec1/stra13/sharp2 in colon carcinoma: its antagonizing role in serum deprivation-induced apoptosis and selective inhibition of procaspase activation. Biochem J. 367:413–422. 2002. View Article : Google Scholar
29	Wang W, Reiser-Erkan C, Michalski CW, et al: Hypoxia inducible BHLHB2 is a novel and independent prognostic marker in pancreatic ductal adenocarcinoma. Biochem Biophys Res Commun. 401:422–428. 2010. View Article : Google Scholar : PubMed/NCBI
30	Kon N, Hirota T, Kawamoto T, et al: Activation of TGF-beta/ activin signalling resets the circadian clock through rapid induction of Dec1 transcripts. Nat Cell Biol. 10:1463–1469. 2008. View Article : Google Scholar : PubMed/NCBI
31	Nicolás FJ and Hill CS: Attenuation of the TGF-beta-Smad signaling pathway in pancreatic tumor cells confers resistance to TGF-beta-induced growth arrest. Oncogene. 22:3698–3711. 2003.PubMed/NCBI
32	Sato F, Kawamoto T, Fujimoto K, et al: Functional analysis of the basic helix-loop-helix transcription factor DEC1 in circadian regulation. Interaction with BMAL1. Eur J Biochem. 271:4409–4419. 2004. View Article : Google Scholar : PubMed/NCBI
33	Kawamoto T, Noshiro M, Sato F, et al: A novel autofeedback loop of Dec1 transcription involved in circadian rhythm regulation. Biochem Biophys Res Commun. 313:117–124. 2004. View Article : Google Scholar : PubMed/NCBI
34	Bloom BB, Humphries DE, Kuang PP, et al: Structure and expression of the promoter for the R4/ALK5 human type I transforming growth factor-beta receptor: regulation by TGF-beta. Biochim Biophys Acta. 1312:243–248. 1996. View Article : Google Scholar : PubMed/NCBI
35	Mulder KM: Role of ras and Mapks in TGF-beta signaling. Cytokine Growth Factor Rev. 11:23–35. 2000. View Article : Google Scholar : PubMed/NCBI
36	Michl P, Barth C, Buchholz M, et al: Claudin-4 expression decreases invasiveness and metastatic potential of pancreatic cancer. Cancer Res. 63:6265–6271. 2003.PubMed/NCBI
37	Turksen K and Troy TC: Junctions gone bad: Claudins and loss of the barrier in cancer. Biochim Biophys Acta. 1816:73–79. 2011.PubMed/NCBI
38	Berx G and Van Roy F: The E-cadherin/catenin complex: an important gatekeeper in breast cancer tumorigenesis and malignant progression. Breast Cancer Res. 3:289–293. 2001. View Article : Google Scholar : PubMed/NCBI
39	Wells A, Yates C and Shepard CR: E-cadherin as an indicator of mesenchymal to epithelial reverting transitions during the metastatic seeding of disseminated carcinomas. Clin Exp Metastasis. 25:621–628. 2008. View Article : Google Scholar : PubMed/NCBI
40	Martínez-Estrada OM, Cullerés A, Soriano FX, et al: The transcription factors Slug and Snail act as repressors of Claudin-1 expression in epithelial cells. Biochem J. 394:449–457. 2006.PubMed/NCBI
41	Batlle E, Sancho E, Francí C, et al: The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol. 2:84–89. 2002. View Article : Google Scholar : PubMed/NCBI
42	Reka AK, Kurapati H, Narala VR, et al: Peroxisome proliferator-activated receptor-gamma activation inhibits tumor metastasis by antagonizing Smad3-mediated epithelial-mesenchymal transition. Mol Cancer Ther. 9:3221–3232. 2010. View Article : Google Scholar