R-spondin 2 promotes proliferation and migration via the Wnt/β-catenin pathway in human hepatocellular carcinoma

Yin,Xinguang; Yi,Huixing; Wang,Linlin; Wu,Wanxin; Wu,Xiaojun; Yu,Linghua

doi:10.3892/ol.2017.6339

R-spondin 2 promotes proliferation and migration via the Wnt/β-catenin pathway in human hepatocellular carcinoma

Authors:
- Xinguang Yin
- Huixing Yi
- Linlin Wang
- Wanxin Wu
- Xiaojun Wu
- Linghua Yu
View Affiliations

Affiliations: Centre for Gastroenterology and Hepatology, The First Affiliated Hospital of Jiaxing College, Jiaxing, Zhejiang 314001, P.R. China, Intensive Care Unit, The Second Affiliated Hospital of Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China, Department of Basic Medicine Sciences, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310058, P.R. China, Deparment of Pathology, The First Affiliated Hospital of Jiaxing College, Jiaxing, Zhejiang 314001, P.R. China
Published online on: June 7, 2017 https://doi.org/10.3892/ol.2017.6339
Pages: 1757-1765

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hepatocellular carcinoma (HCC) is a leading cause of malignant disease-associated mortality, particularly in China. The RSPO2 (R‑spondin 2) gene is evolutionarily conserved in vertebrates and is involved in developmental and physiological processes. Importantly, RSPO2 has been reported to be associated with colon cancer and potentiate the Wnt/β‑catenin signaling pathway. In the present study, enhanced expression of RSPO2 in HCC was observed using tissue microarray. Similarly, the expression level of RSPO2 was higher in HepG2, Huh7 and Hep3B cells but lower in Bel7404 and QGY7703 cells compared with human normal QSG7701 liver cells. Subsequently, gain‑of‑function studies indicated that RSPO2 promotes the proliferation and migration of QGY7703 cells based on lentivirus‑based gene delivery. Furthermore, it was revealed that p21 and leptin, rather than vascular endothelial growth factor‑A, are involved in the function of RSPO2 in QGY7703 cells. Particularly, the signal transducer and activator of transcription 3 (STAT3) and Wnt/β‑catenin signaling pathways are involved in this process. Overexpression of RSPO2 resulted in the elevated expression of phosphorylated STAT3, β‑catenin and c‑Myc. Therefore, the present study is beneficial to the understanding of RSPO2‑involved liver cancer transformation and drug discovery.

View Figures

View References

1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	Liaw YF, Kao JH, Piratvisuth T, Chan HL, Chien RN, Liu CJ, Gane E, Locarnini S, Lim SG, Han KH, et al: Asian-Pacific consensus statement on the management of chronic hepatitis B: A 2012 update. Hepatol Int. 6:531–561. 2012. View Article : Google Scholar : PubMed/NCBI
3	Maluccio M and Covey A: Recent progress in understanding, diagnosing, and treating hepatocellular carcinoma. CA Cancer J Clin. 62:394–399. 2012. View Article : Google Scholar : PubMed/NCBI
4	Zhu GQ, Shi KQ, Yu HJ, He SY, Braddock M, Zhou MT, Chen YP and Zheng MH: Optimal adjuvant therapy for resected hepatocellular carcinoma: A systematic review with network meta-analysis. Oncotarget. 6:18151–18161. 2015. View Article : Google Scholar : PubMed/NCBI
5	de Lau WB, Snel B and Clevers HC: The R-spondin protein family. Genome Biol. 13:2422012. View Article : Google Scholar : PubMed/NCBI
6	Nam JS, Turcotte TJ and Yoon JK: Dynamic expression of R-spondin family genes in mouse development. Gene Expr Patterns. 7:306–312. 2007. View Article : Google Scholar : PubMed/NCBI
7	Parma P, Radi O, Vidal V, Chaboissier MC, Dellambra E, Valentini S, Guerra L, Schedl A and Camerino G: R-spondin1 is essential in sex determination, skin differentiation and malignancy. Nat Genet. 38:1304–1309. 2006. View Article : Google Scholar : PubMed/NCBI
8	Bell SM, Schreiner CM, Wert SE, Mucenski ML, Scott WJ and Whitsett JA: R-spondin 2 is required for normal laryngeal-tracheal, lung and limb morphogenesis. Development. 135:1049–1058. 2008. View Article : Google Scholar : PubMed/NCBI
9	Aoki M, Mieda M, Ikeda T, Hamada Y, Nakamura H and Okamoto H: R-spondin3 is required for mouse placental development. Dev Biol. 301:218–226. 2007. View Article : Google Scholar : PubMed/NCBI
10	Khan TN, Klar J, Nawaz S, Jameel M, Tariq M, Malik NA, Baig SM and Dahl N: Novel missense mutation in the RSPO4 gene in congenital hyponychia and evidence for a polymorphic initiation codon (p.M1I). BMC Med Genet. 13:1202012. View Article : Google Scholar : PubMed/NCBI
11	Lowther W, Wiley K, Smith GH and Callahan R: A new common integration site, Int7, for the mouse mammary tumor virus in mouse mammary tumors identifies a gene whose product has furin-like and thrombospondin-like sequences. J Virol. 79:10093–10096. 2005. View Article : Google Scholar : PubMed/NCBI
12	Theodorou V, Kimm MA, Boer M, Wessels L, Theelen W, Jonkers J and Hilkens J: MMTV insertional mutagenesis identifies genes, gene families and pathways involved in mammary cancer. Nat Genet. 39:759–769. 2007. View Article : Google Scholar : PubMed/NCBI
13	Kim KA, Kakitani M, Zhao J, Oshima T, Tang T, Binnerts M, Liu Y, Boyle B, Park E, Emtage P, et al: Mitogenic influence of human R-spondin1 on the intestinal epithelium. Science. 309:1256–1259. 2005. View Article : Google Scholar : PubMed/NCBI
14	Seshagiri S, Stawiski EW, Durinck S, Modrusan Z, Storm EE, Conboy CB, Chaudhuri S, Guan Y, Janakiraman V, Jaiswal BS, et al: Recurrent R-spondin fusions in colon cancer. Nature. 488:660–664. 2012. View Article : Google Scholar : PubMed/NCBI
15	Shinmura K, Kahyo T, Kato H, Igarashi H, Matsuura S, Nakamura S, Kurachi K, Nakamura T, Ogawa H, Funai K, et al: RSPO fusion transcripts in colorectal cancer in Japanese population. Mol Biol Rep. 41:5375–5384. 2014. View Article : Google Scholar : PubMed/NCBI
16	Kim KA, Wagle M, Tran K, Zhan X, Dixon MA, Liu S, Gros D, Korver W, Yonkovich S, Tomasevic N, et al: R-Spondins family members regulate the Wnt pathway by a common mechanism. Mol Biol Cell. 19:2588–2596. 2008. View Article : Google Scholar : PubMed/NCBI
17	Huch M, Dorrell C, Boj SF, van Es JH, Li VS, van de Wetering M, Sato T, Hamer K, Sasaki N, Finegold MJ, et al: In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature. 494:247–250. 2013. View Article : Google Scholar : PubMed/NCBI
18	Carmon KS, Gong X, Yi J, Thomas A and Liu Q: RSPO-LGR4 functions via IQGAP1 to potentiate Wnt signaling. Proc Natl Acad Sci USA. 111:pp. E1221–E1229. 2014; View Article : Google Scholar : PubMed/NCBI
19	Logan CY and Nusse R: The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 20:781–810. 2004. View Article : Google Scholar : PubMed/NCBI
20	Clevers H: Wnt/beta-catenin signaling in development and disease. Cell. 127:469–480. 2006. View Article : Google Scholar : PubMed/NCBI
21	Monga SP: β-Catenin signaling and roles in liver homeostasis, injury and tumorigenesis. Gastroenterology. 148:1294–1310. 2015. View Article : Google Scholar : PubMed/NCBI
22	Stewart DJ: Wnt signaling pathway in non-small cell lung cancer. J Natl Cancer Inst. 106:djt3562014. View Article : Google Scholar : PubMed/NCBI
23	Gregorieff A, Liu Y, Inanlou MR, Khomchuk Y and Wrana JL: Yap-dependent reprogramming of Lgr5(+) stem cells drives intestinal regeneration and cancer. Nature. 526:715–718. 2015. View Article : Google Scholar : PubMed/NCBI
24	Hu T and Li C: Convergence between Wnt-β-catenin and EGFR signaling in cancer. Mol Cancer. 9:2362010. View Article : Google Scholar : PubMed/NCBI
25	Ahn SM, Jang SJ, Shim JH, Kim D, Hong SM, Sung CO, Baek D, Haq F, Ansari AA, Lee SY, et al: Genomic portrait of resectable hepatocellular carcinomas: Implications of RB1 and FGF19 aberrations for patient stratification. Hepatology. 60:1972–1982. 2014. View Article : Google Scholar : PubMed/NCBI
26	Kononen J, Bubendorf L, Kallioniemi A, Bärlund M, Schraml P, Leighton S, Torhorst J, Mihatsch MJ, Sauter G and Kallioniemi OP: Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med. 4:844–847. 1998. View Article : Google Scholar : PubMed/NCBI
27	Boseman FT, Carneiro F, Hruban RH and Theise ND: Tumours of the liver and intrahepatic bile ductsWorld Health Organization Classification of Tumours of the Digestive System. 4th. IARC; Lyon: pp. 195–262. 2010
28	Kang N, Gores GJ and Shah VH: Hepatic stellate cells: Partners in crime for liver metastases? Hepatology. 54:707–713. 2011. View Article : Google Scholar : PubMed/NCBI
29	Coulouarn C and Clément B: Stellate cells and the development of liver cancer: Therapeutic potential of targeting the stroma. J Hepatol. 60:1306–1309. 2014. View Article : Google Scholar : PubMed/NCBI
30	Xinguang Y, Huixing Y, Xiaowei W, Xiaojun W and Linghua Y: R-spondin1 arguments hepatic fibrogenesis in vivo and in vitro. J Surg Res. 193:598–605. 2015. View Article : Google Scholar : PubMed/NCBI
31	Yin X, Yi H, Wu W, Shu J, Wu X and Yu L: R-spondin2 activates hepatic stellate cells and promotes liver fibrosis. Dig Dis Sci. 59:2452–3961. 2014. View Article : Google Scholar : PubMed/NCBI
32	Chartier C, Raval J, Axelrod F, Bond C, Cain J, Dee-Hoskins C, Ma S, Fischer MM, Shah J, Wei J, et al: Therapeutic targeting of tumor-derived R-spondin attenuates β-catenin signaling and tumorigenesis in multiple cancer types. Cancer Res. 76:713–722. 2016. View Article : Google Scholar : PubMed/NCBI