Top

Journal of Experimental & Clinical Cancer Research

Published in:

Open Access 01-12-2018 | Research

Cyclin G2 suppresses Wnt/β-catenin signaling and inhibits gastric cancer cell growth and migration through Dapper1

Authors: Jinlan Gao, Chenyang Zhao, Qi Liu, Xiaoyu Hou, Sen Li, Xuesha Xing, Chunhua Yang, Yang Luo

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2018

Abstract

Background

Gastric cancer is one of the most common malignant tumors. Cyclin G2 has been shown to be associated with the development of multiple types of tumors, but its underlying mechanisms in gastric tumors is not well-understood. The aim of this study is to investigate the role and the underlying mechanisms of cyclin G2 on Wnt/β-catenin signaling in gastric cancer.

Methods

Real-time PCR, immunohistochemistry and in silico assay were used to determine the expression of cyclin G2 in gastric cancer. TCGA datasets were used to evaluate the association between cyclin G2 expression and the prognostic landscape of gastric cancers. The effects of ectopic and endogenous cyclin G2 on the proliferation and migration of gastric cancer cells were assessed using the MTS assay, colony formation assay, cell cycle assay, wound healing assay and transwell assay. Moreover, a xenograft model and a metastasis model of nude mice was used to determine the influence of cyclin G2 on gastric tumor growth and migration in vivo. The effects of cyclin G2 expression on Wnt/β-catenin signaling were explored using a TOPFlash luciferase reporter assay, and the molecular mechanisms involved were investigated using immunoblots assay, yeast two-hybrid screening, immunoprecipitation and Duolink in situ PLA. Ccng2^−/− mice were generated to further confirm the inhibitory effect of cyclin G2 on Wnt/β-catenin signaling in vivo. Furthermore, GSK-3β inhibitors were utilized to explore the role of Wnt/β-catenin signaling in the suppression effect of cyclin G2 on gastric cancer cell proliferation and migration.

Results

We found that cyclin G2 levels were decreased in gastric cancer tissues and were associated with tumor size, migration and poor differentiation status. Moreover, overexpression of cyclin G2 attenuated tumor growth and metastasis both in vitro and in vivo. Dpr1 was identified as a cyclin G2-interacting protein which was required for the cyclin G2-mediated inhibition of β-catenin expression. Mechanically, cyclin G2 impacted the activity of CKI to phosphorylate Dpr1, which has been proved to be a protein that acts as a suppressor of Wnt/β-catenin signaling when unphosphorylated. Furthermore, GSK-3β inhibitors abolished the cyclin G2-induced suppression of cell proliferation and migration.

Conclusions

This study demonstrates that cyclin G2 suppresses Wnt/β-catenin signaling and inhibits gastric cancer cell growth and migration through Dapper1.

Available only for authorised users

Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-tieulent J, Jemal A. Global Cancer. Statistics, 2012. CA a cancer J Clin. 2015;65:87–108.CrossRef

Yu J, Liang Q, Wang J, Wang K, Gao J, Zhang J, et al. REC8 functions as a tumor suppressor and is epigenetically downregulated in gastric cancer, especially in EBV-positive subtype. Oncogene. 2017;36:182–93.CrossRef

Chiurillo MA. Role of the Wnt/β-catenin pathway in gastric cancer: an in-depth literature review. World J Exp Med. 2015;5:84–102.CrossRef

Clements WM, Wang J, Sarnaik A, Kim OJ, MacDonald J, Fenoglio-Preiser C, et al. Beta-catenin mutation is a frequent cause of Wnt pathway activation in gastric cancer. Cancer Res. 2002;62:3503–6.PubMed

Horne MC, Goolsby GL, Donaldson KL, Tran D, Neubauer M, Wahl AF. Cyclin G1 and cyclin G2 comprise a new family of cyclins with contrasting tissue-specific and cell cycle-regulated expression. J Biol Chem. 1996;271:6050–61.CrossRef

Martínez-Gac L, Marqués M, García Z, Campanero MR, Carrera AC. Control of cyclin G2 mRNA expression by forkhead transcription factors: novel mechanism for cell cycle control by phosphoinositide 3-kinase and forkhead. Mol Cell Biol. 2004;24:2181–9.CrossRef

Choi MG, Noh JH, An JY, Hong SK, Park SB, Baik YH, et al. Expression levels of cyclin G2, but not cyclin E, correlate with gastric cancer progression. J Surg Res. 2009;157:168–74.CrossRef

Gao J, Liu Q, Liu X, Zhang Y, Wang S, Luo Y. Mutation analysis of the negative regulator cyclin G2 in gastric cancer. Afi J Bio technol. 2011;10:14618–24.

Bergqvist M, Brattström D, Brodin D, Lindkvist A, Dreilich M, Wagenius G. Genes associated with telomerase activity levels in esophageal carcinoma cell lines. Dis Esphagos. 2006;19:20–3.CrossRef

10.

Fu G, Peng C. Nodal enhances the activity of FoxO3a and its synergistic interaction with Smads to regulate cyclin G2 transcription in ovarian cancer cells. Oncogene. 2011;30:3953–66.CrossRef

11.

Zvibel I, Wagner A, Pasmanik-Chor M, Varol C, Oron-Karni V, Santo EM, et al. Transcriptional profiling identifies genes induced by hepatocyte-derived extracellular matrix in metastatic human colorectal cancer cell lines. Clin Exp Metastasis. 2013;30:189–200.CrossRef

12.

Li W, Liu G, Yu F, Xiang X, Lu Y, Xiao H, et al. CCNG2 suppressor biological effects on thyroid cancer cell through promotion of CDK2 degradation. Asian Pacific J Cancer Prev. 2013;14:6165–71.

13.

Shen Y, Takahashi M, Byun HM, Link A, Sharma N, Balaguer F, et al. Boswellic acid induces epigenetic alterations by modulating DNA methylation in colorectal cancer cells. Cancer Biol Ther. 2012;13:7542–52.CrossRef

14.

Kim Y, Shintani S, Kohno Y, Zhang R, Wong DT. Cyclin G2 dysregulation in human Oral Cancer. Cancer Res. 2004;64:8980–6.CrossRef

15.

Cui DW, Cheng YJ, Jing SW, Sun GG. Effect of cyclin G2 on proliferative ability of prostate cancer PC-3 cell. Tumor Biol. 2014;35:3017–24.CrossRef

16.

Clevers H. Review Wnt/β-catenin signaling in development and disease. Cell. 2006;127:469–80.

17.

Korinek V, Barker N, Willert K, Molenaar M, Roose J, Wagenaar G, et al. Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. Mol Cell Biol. 1998;18:1248–56.CrossRef

18.

Li Q, Ye L, Guo W, Wang M, Huang S, Peng X. PHF21B overexpression promotes cancer stem cell-like traits in prostate cancer cells by activating the Wnt/β-catenin signaling pathway. J Exp Clin Canc Res. 2017;36:85.CrossRef

19.

Wu D, Pan W. GSK3 : a multifaceted kinase in Wnt signaling. Trends Biochem Sci. 2009;35:161–8.CrossRef

20.

Liu C, Li Y, Semenov M, Han C, Baeg GH, Tan Y, et al. Control of B-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell. 2002;108:837–47.CrossRef

21.

MacDonald BT, Tamai K, He X. Wnt β-catenin signaling components mechanisms and disease. Development. 2009;135:367–75.

22.

Zeng X, Huang H, Tamai K, Zhang X, Harada Y, Yokota C, et al. Initiation of Wnt signaling: control of Wnt coreceptor Lrp6 phosphorylation/activation via frizzled, dishevelled and axin functions. Development. 2008;135:367–75.CrossRef

23.

Gao X, Wen J, Zhang L, Li X, Ning Y, Meng A, et al. Dapper1 is a nucleocytoplasmic shuttling protein that negatively modulates Wnt signaling in the nucleus. J Biol Chem. 2008;283:35679–88.CrossRef

24.

Chen H, Liu L, Ma B, Ma TM, Hou JJ, Xie GM, et al. Protein kinase A-mediated 14-3-3 association impedes human Dapper1 to promote Dishevelled degradation. J Biol Chem. 2011;286:14870–80.

25.

Zhang L, Gao X, Wen J, Ning Y, Chen YG. Dapper 1 antagonizes Wnt signaling by promoting dishevelled degradation. J Biol Chem. 2006;281:8607–12.

26.

Gaur T, Lengner CJ, Hovhannisyan H, Bhat RA, Bodine PVN, Komm BS, et al. Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. J Biol Chem. 2005;280:33132–40.

27.

Shtutman M, Zhurinsky J, Simcha I, Albanese C, D’Amico M, Pestell R, et al. The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci U S A. 1999;96:5522–7.

28.

Grabsch HI, Tan P. Gastric cancer pathology and underlying molecular mechanisms. Dig Surg. 2013;30:150–8.

29.

Yu Y, Yan W, Liu X, Jia Y, Cao B, Yu Y, et al. DACT2 is frequently methylated in human gastric cancer and methylation of DACT2 activated Wnt signaling. Am J Cancer Res. 2014;4:710–24.

30.

Huang J, Xiao D, Li G, Ma J, Chen P, Yuan W, et al. EphA2 promotes epithelial-mesenchymal transition through the Wnt/beta-catenin pathway in gastric cancer cells. Oncogene. 2014;33:2737–47.

31.

Yue Z, Fan Y, Jing J, Zheng F, Ping T, Ling Y et al. RACK1 suppresses gastric tumorigenesis by stabilizing the β-Catenin destruction complex. Gastroenterlogy 2012; 142: 812–23.

32.

Gao J, Liu Q, Liu X, Ji C, Qu S, Wang S et al. Cyclin G2 suppresses estrogen-mediated osteogenesis through inhibition of Wnt/β-catenin signaling. PLoS One 2014; 9: e89884.

33.

Rhodes DR, Yu J, Shanker K, Deshpande N, Varambally R, Ghosh D, Barrette T, Pandey A, Chinnaiyan AM, et al. ONCOMINE: a Cancer microarray database and integrated data-mining platform. Neoplasia. 2004;6(1):1–6.

34.

Ooi CH, Ivanova T, Wu J, Lee M, Tan IB, Tao J, et al. Oncogenic pathway combinations predict clinical prognosis in gastric cancer. PLoS Genet. 2009;5(10):e1000676.

35.

D'Errico M, de Rinaldis E, Blasi MF, Viti V, Falchetti M, Calcagnile A, et al. Genome-wide expression profile of sporadic gastric cancers with microsatellite instability. Eur J Cancer. 2009;45(3):461–9.

36.

Forster S, Gretschel S, Jons T, Yashiro M, Kemmner W. THBS4, a novel stromal molecule of diffuse-type gastric adenocarcinomas, identified by transcriptome-wide expression profiling. Mod Pathol. 2011;24(10):1390–403.

37.

Chen X, Leung SY, Yuen ST, Chu KM, Ji J, Li R. Variation in gene expression patterns in human gastric cancers. Mol Biol Cell. 2003;14(8):3208–15.

38.

Cho JY, Lim JY, Cheong JH, Park YY, Yoon SL, Kim SM, et al. Gene expression signature-based prognostic risk score in gastric cancer. Clin Cancer Res. 2011;17(7):1850–7.

39.

Liu Q, Li X, Li S, Qu S, Wang Y, Tang Q, et al. Three novel mutations of APC gene in Chinese patients with familial adenomatous polyposis. Tumor Biol. 2016;37:11421–7.

40.

Xu G, Bernaudo S, Fu G, Lee DY, Yang BB, Peng C. Cyclin G2 is degraded through the ubiquitin-proteasome pathway and mediates the antiproliferative effect of activin receptor-like kinase 7. Mol Biol Cell. 2008;9:4968–79.

41.

Veeman MT, Ajamete Kaykas SHL, Moon RT. Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, regulates gastrulation movements. Curr Biol. 2003;13:680–5.

42.

Spandidos A, Wang X, Wang H, Seed B. PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification. Nucleic Acids Res. 2009;38:792–9.

43.

Liu Q, Liu X, Gao J, Shi X, Hu X, Wang S, et al. Overexpression of DOC-1R inhibits cell cycle G1/S transition by repressing CDK2 expression and activation. Int J Biol Sci. 2013;9:541–9.

44.

Clevers H, Nusse R. Wnt/β-catenin signaling and disease. Cell. 2012;149:1192–205.

45.

Teran E, Branscomb AD, Seeling JM. Dpr acts as a molecular switch, inhibiting Wnt signaling when unphosphorylated, but promoting Wnt signaling when phosphorylated by casein kinase Idelta/epsilon. PLoS One. 2009;4:e5522.

46.

Sineva GS, Pospelov VA. Inhibition of GSK3beta enhances both adhesive and signalling activities of beta-catenin in mouse embryonic stem cells. Biol Cell. 2010;102:549–60.

47.

Caca K, Kolligs FT, Ji X, Hayes M, Qian J, Yahanda A, et al. β- and γ-catenin mutations, but not E-cadherin inactivation, underlie T-cell factor/lymphoid enhancer factor transcriptional deregulation in gastric and pancreatic cancer. Cell Growth Differ. 1999;10:369–76.

48.

Bernaudo S, Salem M, Qi X, Zhou W, Zhang C, Yang W, et al. Cyclin G2 inhibits epithelial-to-mesenchymal transition by disrupting Wnt/β-catenin signaling. Oncogene. 2016;35:4816–27.

Title: Cyclin G2 suppresses Wnt/β-catenin signaling and inhibits gastric cancer cell growth and migration through Dapper1
Authors: Jinlan Gao
Chenyang Zhao
Qi Liu
Xiaoyu Hou
Sen Li
Xuesha Xing
Chunhua Yang
Yang Luo
Publication date: 01-12-2018
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2018
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-018-0973-2

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2018

Advances in circular RNAs and their roles in breast Cancer

Tetraspanin 1 promotes epithelial-to-mesenchymal transition and metastasis of cholangiocarcinoma via PI3K/AKT signaling

Dacomitinib potentiates the efficacy of conventional chemotherapeutic agents via inhibiting the drug efflux function of ABCG2 in vitro and in vivo

Hepatocellular carcinoma-derived exosomal miRNA-21 contributes to tumor progression by converting hepatocyte stellate cells to cancer-associated fibroblasts

Anti-tumor effects of ONC201 in combination with VEGF-inhibitors significantly impacts colorectal cancer growth and survival in vivo through complementary non-overlapping mechanisms

Liquid biopsy in mice bearing colorectal carcinoma xenografts: gateways regulating the levels of circulating tumor DNA (ctDNA) and miRNA (ctmiRNA)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer