ASCL2 expression contributes to gastric tumor migration and invasion by downregulating miR223 and inducing EMT

Zuo,Qingsong; Wang,Jie; Chen,Chao; Zhang,Yong; Feng,Dian‑Xu; Zhao,Ronghua; Chen,Teng

doi:10.3892/mmr.2018.9363

ASCL2 expression contributes to gastric tumor migration and invasion by downregulating miR223 and inducing EMT

Authors:
- Qingsong Zuo
- Jie Wang
- Chao Chen
- Yong Zhang
- Dian‑Xu Feng
- Ronghua Zhao
- Teng Chen
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Affiliations: Department of General Surgery, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, P.R. China
Published online on: August 8, 2018 https://doi.org/10.3892/mmr.2018.9363
Pages: 3751-3759
Copyright: © Zuo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Achaete‑scute homolog 2 (ASCL2), a basic helix‑loop‑helix transcription factor, serves an essential role in the maintenance of adult intestinal stem cells and the growth of gastric cancer (GC). However, the function of ASCL2 in the metastasis of GC is poorly understood. The present study aimed to evaluate the effect of ASCL2 expression on gastric tumor metastasis. ASCL2 protein expression was detected in 32 cases of gastric metastasis and its relevant primary tumors using western blotting and immunohistochemistry. The data suggested that the expression of ASCL2 was highest in metastatic tumors, among adjacent normal tissues, primary gastric tumors and gastric metastatic tumors. Furthermore, ASCL2‑overexpressing GC cell lines MKN1‑ASCL2 and SNU16‑ASCL2 were established. An in vitro assay suggested that microRNA 223 (miR223) expression was downregulated following ASCL2 overexpression, and that the expression of the epithelium‑associated protein E‑cadherin was significantly decreased, while a series of mesenchyme‑associated proteins, including zinc finger E‑box‑binding homeobox 1 (Zeb‑1), twist‑related protein 1, integrin, vimentin, 72 kDa type IV collagenase and matrix metalloproteinase‑9 were upregulated in ASCL2‑overexpressing cells. Overexpression of miR223 attenuated the epithelial‑mesenchymal transition (EMT)‑promoting effect induced by ASCL2 expression. In addition, the results of the chromatin immunoprecipitation and luciferase reporter gene assays indicated that ASCL2 was able to interact with the promoter of pre‑miR223, and to inhibit the maturation of miR223, which may interact with the 3' untranslated region of Zeb‑1 and inhibit EMT in tumor cells. The results of the present study demonstrated that ASCL2 was able to downregulate the expression level of miR223, contribute to EMT and promote gastric tumor metastasis, which indicated that ASCL2 may serve as a therapeutic target in the treatment of GC.

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1	Kwon OH, Park JL, Baek SJ, Noh SM, Song KS, Kim SY and Kim YS: Aberrant upregulation of ASCL2 by promoter demethylation promotes the growth and resistance to 5-fluorouracil of gastric cancer cells. Cancer Sci. 104:391–397. 2013. View Article : Google Scholar : PubMed/NCBI
2	Pisani P, Parkin DM, Bray F and Ferlay J: Erratum: Estimates of the worldwide mortality from 25 cancers in 1990. Int J Cancer, 83, 18–29 (1999). Int J Cancer. 83:870–873. 1999. View Article : Google Scholar : PubMed/NCBI
3	Yuasa Y: Control of gut differentiation and intestinal-type gastric carcinogenesis. Nat Rev Cancer. 3:592–600. 2003. View Article : Google Scholar : PubMed/NCBI
4	Massari ME and Murre C: Helix-loop-helix proteins: Regulators of transcription in eucaryotic organisms. Mol Cell Biol. 20:429–440. 2000. View Article : Google Scholar : PubMed/NCBI
5	Miyamoto T, Jinno Y, Sasaki T, Ikeda Y, Masuzaki H, Niikawa N and Ishikawa M: Genomic cloning and localization to chromosome 11p15.5 of the human achaete-scute homolog 2 (ASCL2). Cytogenet Cell Genet. 73:312–314. 1996. View Article : Google Scholar : PubMed/NCBI
6	van der Flier LG, van Gijn ME, Hatzis P, Kujala P, Haegebarth A, Stange DE, Begthel H, van den Born M, Guryev V, Oving I, et al: Transcription factor achaete scute-like 2 controls intestinal stem cell fate. Cell. 136:903–912. 2009. View Article : Google Scholar : PubMed/NCBI
7	Jubb AM, Chalasani S, Frantz GD, Smits R, Grabsch HI, Kavi V, Maughan NJ, Hillan KJ, Quirke P and Koeppen H: Achaete-scute like 2 (ascl2) is a target of Wnt signalling and is upregulated in intestinal neoplasia. Oncogene. 25:3445–3457. 2006. View Article : Google Scholar : PubMed/NCBI
8	Zhu R, Yang Y, Tian Y, Bai J, Zhang X, Li X, Peng Z, He Y, Chen L, Pan Q, et al: Ascl2 knockdown results in tumor growth arrest by miRNA-302b-related inhibition of colon cancer progenitor cells. PLoS One. 7:e321702012. View Article : Google Scholar : PubMed/NCBI
9	Nicoloso MS, Spizzo R, Shimizu M, Rossi S and Calin GA: MicroRNAs-the micro steering wheel of tumour metastases. Nat Rev Cancer. 9:293–302. 2009. View Article : Google Scholar : PubMed/NCBI
10	Gal H, Pandi G, Kanner AA, Ram Z, Lithwick-Yanai G, Amariglio N, Rechavi G and Givol D: MIR-451 and Imatinib mesylate inhibit tumor growth of Glioblastoma stem cells. Biochem Biophys Res Commun. 376:86–90. 2008. View Article : Google Scholar : PubMed/NCBI
11	Lee NS, Kim JS, Cho WJ, Lee MR, Steiner R, Gompers A, Ling D, Zhang J, Strom P, Behlke M, et al: miR-302b maintains ‘stemness’ of human embryonal carcinoma cells by post-transcriptional regulation of Cyclin D2 expression. Biochem Biophys Res Commun. 377:434–440. 2008. View Article : Google Scholar : PubMed/NCBI
12	Yu F, Yao H, Zhu P, Zhang X, Pan Q, Gong C, Huang Y, Hu X, Su F, Lieberman J and Song E: let-7 regulates self renewal and tumorigenicity of breast cancer cells. Cell. 131:1109–1123. 2007. View Article : Google Scholar : PubMed/NCBI
13	Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T and Thun MJ: Cancer statistics, 2008. CA Cancer J Clin. 58:71–96. 2008. View Article : Google Scholar : PubMed/NCBI
14	Tsujimoto H, Ono S, Ichikura T, Matsumoto Y, Yamamoto J and Hase K: Roles of inflammatory cytokines in the progression of gastric cancer: Friends or foes? Gastric Cancer. 13:212–221. 2010. View Article : Google Scholar : PubMed/NCBI
15	Tian Y, Pan Q, Shang Y, Zhu R, Ye J, Liu Y, Zhong X, Li S, He Y, Chen L, et al: MicroRNA-200 (miR-200) cluster regulation by achaete scute-like 2 (Ascl2): Impact on the epithelial-mesenchymal transition in colon cancer cells. J Biol Chem. 289:36101–36115. 2014. View Article : Google Scholar : PubMed/NCBI
16	Solt LA, Griffin PR and Burris TP: Ligand regulation of retinoic acid receptor-related orphan receptors: Implications for development of novel therapeutics. Curr Opin Lipidol. 21:204–211. 2010. View Article : Google Scholar : PubMed/NCBI
17	Becker-Andre M, Andre E and DeLamarter JF: Identification of nuclear receptor mRNAs by RT-PCR amplification of conserved zinc-finger motif sequences. Biochem Biophys Res Commun. 194:1371–1379. 1993. View Article : Google Scholar : PubMed/NCBI
18	Schuijers J, Junker JP, Mokry M, Hatzis P, Koo BK, Sasselli V, Van der Flier LG, Cuppen E, van Oudenaarden A and Clevers H: Ascl2 acts as an R-spondin/Wnt-responsive switch to control stemness in intestinal crypts. Cell Stem Cell. 16:158–170. 2015. View Article : Google Scholar : PubMed/NCBI
19	Carlberg C, Hooft van Huijsduijnen R, Staple JK, DeLamarter JF and Becker-André M: RZRs, a new family of retinoid-related orphan receptors that function as both monomers and homodimers. Mol Endocrinol. 8:757–770. 1994. View Article : Google Scholar : PubMed/NCBI
20	Jetten AM, Kurebayashi S and Ueda E: The ROR nuclear orphan receptor subfamily: Critical regulators of multiple biological processes. Prog Nucleic Acid Res Mol Biol. 69:205–247. 2001. View Article : Google Scholar : PubMed/NCBI
21	Gawlas K and Stunnenberg HG: Differential transcription of the orphan receptor RORbeta in nuclear extracts derived from Neuro2A and HeLa cells. Nucleic Acids Res. 29:3424–3432. 2001. View Article : Google Scholar : PubMed/NCBI
22	Chen CZ, Li L, Lodish HF and Bartel DP: MicroRNAs modulate hematopoietic lineage differentiation. Science. 303:83–86. 2004. View Article : Google Scholar : PubMed/NCBI
23	Stamatopoulos B, Meuleman N, Haibe-Kains B, Saussoy P, Van Den Neste E, Michaux L, Heimann P, Martiat P, Bron D and Lagneaux L: microRNA-29c and microRNA-223 down-regulation has in vivo significance in chronic lymphocytic leukemia and improves disease risk stratification. Blood. 113:5237–5245. 2009. View Article : Google Scholar : PubMed/NCBI
24	Mi S, Lu J, Sun M, Li Z, Zhang H, Neilly MB, Wang Y, Qian Z, Jin J, Zhang Y, et al: MicroRNA expression signatures accurately discriminate acute lymphoblastic leukemia from acute myeloid leukemia. Proc Natl Acad Sci USA. 104:pp. 19971–19976. 2007; View Article : Google Scholar : PubMed/NCBI