Expression of sphingosine‑1‑phosphate receptor 2 is correlated with migration and invasion of human colon cancer cells: A preliminary clinical study

Yan,Junjun; Chen,Yi; Wu,Qibiao; Shao,Le; Zhou,Xiqiao

doi:10.3892/ol.2022.13361

Expression of sphingosine‑1‑phosphate receptor 2 is correlated with migration and invasion of human colon cancer cells: A preliminary clinical study

Authors:
- Junjun Yan
- Yi Chen
- Qibiao Wu
- Le Shao
- Xiqiao Zhou
View Affiliations

Affiliations: Department of Gastroenterology, The First People's Hospital of Jiujiang, Jiujiang, Jiangxi 332000, P.R. China, Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, State Key Laboratory of Quality Research in Chinese Medicines and Faculty of Chinese Medicine, Macau University of Science and Technology (MUST), Taipa, Macau 999078, P.R. China, Center for Medical Research and Innovation, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410000, P.R. China
Published online on: June 1, 2022 https://doi.org/10.3892/ol.2022.13361
Article Number: 241
Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Sphingosine‑1‑phosphate (S1P) is a bioactive phospholipid that serves as a potent mediator of cell proliferation, differentiation and apoptosis by binding to S1P receptors (S1PRs). S1P signalling is involved in the pathogenesis of numerous types of disease, including cancer. To the best of our knowledge, however, little is known about the expression patterns of S1PRs and their role in human colorectal cancer (CRC) cell migration and invasion. The aim of the present study was to investigate the role of S1P signalling in the metastasis of colon cancer cells and the expression of S1PRs in patients with CRC. The protein and mRNA expression levels of S1PRs and sphingosine kinases (SPHKs) in 55 patients with CRC were detected by western blotting (WB), immunohistochemical (IHC) analysis and reverse transcription‑quantitative PCR. The levels of S1P in serum from patients and healthy individuals were quantified by ELISA. S1PRs antagonists JTE013, FTY720 and S1PR2‑small interfering (si)RNA were used to determine the role of S1PR2 in human CRC LOVO and SW480 cell lines. Migration and invasion assays were performed for functional analysis. The levels of S1P in serum were significantly increased in patients with CRC compared with healthy individuals. The relative mRNA expression levels of S1PR2 were significantly downregulated in tumour compared with normal tissue, whereas S1PR1 and SPHK1 were upregulated. WB showed that 58% (32/55 cases) of patients presented downregulated S1PR2 protein expression. IHC analysis indicated that expression of S1PR2 was lower in tumour than in normal tissue in 65.5% (36/55 cases) of patients. Exogenous addition of S1P promoted migration and invasion in the different cell types. S1P stimulated the migration and invasion of SW480 cells. The inhibition of S1PR2 by JTE013 or S1PR2‑siRNA significantly promoted the migration and invasion of SW480 cells, while FTY720 reversed these effects. The present study indicated that expression levels of S1PRs, particularly S1PR2, were associated with migration and invasion of CRC cells. The present findings revealed a novel mechanism by which S1P inhibited tumour cell migration and invasion via a S1PR2‑dependent pathway, suggesting that S1PR2 may be a therapeutic target for treatment of colon cancer.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer Statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar : PubMed/NCBI
2	Van Doorn R, Van Horssen J, Verzijl D, Witte M, Ronken E, Van Het HB, Lakeman K, Dijkstra CD, Van Der Valk P, Reijerkerk A, et al: Sphingosine 1-phosphate receptor 1 and 3 are upregulated in multiple sclerosis lesions. Glia. 58:1465–1476. 2010. View Article : Google Scholar
3	Zhang G, Yang L, Kim GS, Ryan K, Lu S, O'Donnell RK, Spokes K, Shapiro N, Aird WC, Kluk MJ, et al: Critical role of sphingosine-1-phosphate receptor 2 (S1PR2) in acute vascular inflammation. Blood. 122:443–455. 2013. View Article : Google Scholar : PubMed/NCBI
4	Baldari CT: The hematopoietic oncoprotein FOXP1 promotes tumor cell survival in diffuse large B-cell lymphoma by repressing S1PR2 signaling. Blood. 127:1380–1381. 2016. View Article : Google Scholar : PubMed/NCBI
5	Green JA and Cyster JG: S1PR2 links germinal center confinement and growth regulation. Immunol Rev. 247:36–51. 2012. View Article : Google Scholar : PubMed/NCBI
6	Green JA, Suzuki K, Cho B, Willison LD, Palmer D, Allen CD, Schmidt TH, Xu Y, Proia RL, Coughlin SR and Cyster JG: The sphingosine 1-phosphate receptor S1P₂ maintains the homeostasis of germinal center B cells and promotes niche confinement. Nat Immunol. 12:672–680. 2011. View Article : Google Scholar
7	Sánchez DI, González-Fernández B, San-Miguel B, de Urbina JO, Crespo I, González-Gallego J and Tuñón MJ: Melatonin prevents deregulation of the sphingosine kinase/sphingosine 1-phosphate signaling pathway in a mouse model of diethylnitrosamine-induced hepatocellular carcinoma. J Pineal Res. 62:e123692017. View Article : Google Scholar
8	Higashi K, Matsuzaki E, Hashimoto Y, Takahashi-Yanaga F, Takano A, Anan H, Hirata M and Nishimura F: Sphingosine-1-phosphate/S1PR2-mediated signaling triggers Smad1/5/8 phosphorylation and thereby induces Runx2 expression in osteoblasts. Bone. 93:1–11. 2016. View Article : Google Scholar
9	Nagahashi M, Yuza K, Hirose Y, Nakajima M, Ramanathan R, Hait NC, Hylemon PB, Zhou H, Takabe K and Wakai T: The roles of bile acids and sphingosine-1-phosphate signaling in the hepatobiliary diseases. J Lipid Res. 57:1636–1643. 2016. View Article : Google Scholar : PubMed/NCBI
10	Shida D, Inoue S, Yoshida Y, Kodaka A, Tsuji T and Tsuiji M: Sphingosine kinase 1 is upregulated with lysophosphatidic acid receptor 2 in human colorectal cancer. World J Gastroenterol. 22:2503–2511. 2016. View Article : Google Scholar
11	Spiegel S and Milstien S: Sphingosine-1-phosphate: An enigmatic signalling lipid. Nat Rev Mol Cell Bio. 4:397–407. 2003. View Article : Google Scholar
12	Pyne S and Pyne NJ: Sphingosine 1-phosphate signalling in mammalian cells. Biochem J. 349:385–402. 2000. View Article : Google Scholar : PubMed/NCBI
13	Gonzalezcabrera PJ, Brown S, Studer SM and Rosen H: S1P signaling: New therapies and opportunities. F1000 Prime Reports. 6:1092014.PubMed/NCBI
14	Pyne NJ and Pyne S: Sphingosine 1-phosphate and cancer. Nat Rev Cancer. 10:489–503. 2010. View Article : Google Scholar : PubMed/NCBI
15	Pyne NJ, Mcnaughton M, Boomkamp S, Macritchie N, Evangelesti C, Martelli AM, Jiang HR, Ubhi S and Pyne S: Role of sphingosine 1-phosphate receptors, sphingosine kinases and sphingosine in cancer and inflammation. Adv Biol Regul. 60:151–159. 2015. View Article : Google Scholar
16	Sukocheva OA, Furuya H, Ng ML, Friedemann M, Menschikowski M, Tarasov VV, Chubarev VN, Klochkov SG, Neganova ME, Mangoni AA, et al: Sphingosine kinase and sphingosine-1-phosphate receptor signaling pathway in inflammatory gastrointestinal disease and cancers: A novel therapeutic target. Pharmacol Ther. 207:1074642020. View Article : Google Scholar : PubMed/NCBI
17	Okamoto H, Takuwa N, Gonda K, Okazaki H, Chang K, Yatomi Y, Shigematsu H and Takuwa Y: EDG1 is a functional sphingosine-1-phosphate receptor that is linked via a Gi/o to multiple signaling pathways, including phospholipase C activation, Ca2+ mobilization, Ras-mitogen-activated protein kinase activation, and adenylate cyclase inhibition. J Biol Chem. 273:27104–27110. 1998. View Article : Google Scholar
18	Hannun YA and Obeid LM: Principles of bioactive lipid signalling: Lessons from sphingolipids. Nat Rev Mol Cell Bio. 9:139–150. 2008. View Article : Google Scholar
19	Ng ML, Wadham C and Sukocheva OA: The role of sphingolipid signalling in diabetesassociated pathologies (Review). Int J Mol Med. 39:243–252. 2017. View Article : Google Scholar : PubMed/NCBI
20	Arikawa K, Takuwa N, Yamaguchi H, Sugimoto N, Kitayama J, Nagawa H, Takehara K and Takuwa Y: Ligand-dependent inhibition of B16 melanoma cell migration and invasion via endogenous S1P2 G protein-coupled receptor. Requirement of inhibition of cellular RAC activity. J Biol Chem. 278:32841–32851. 2003. View Article : Google Scholar : PubMed/NCBI
21	Liang J, Nagahashi M, Kim EY, Harikumar KB, Yamada A, Huang WC, Hait NC, Allegood JC, Price MM and Avni D: Sphingosine-1-phosphate links persistent STAT3 activation, chronic intestinal inflammation, and development of colitis-associated cancer. Cancer Cell. 23:107–120. 2013. View Article : Google Scholar
22	Du W, Takuwa N, Yoshioka K, Okamoto Y, Gonda K, Sugihara K, Fukamizu A, Asano M and Takuwa Y: S1P(2), the G protein-coupled receptor for sphingosine-1-phosphate, negatively regulates tumor angiogenesis and tumor growth in vivo in mice. Cancer Res. 70:772–781. 2010. View Article : Google Scholar
23	Watson C, Long JS, Orange C, Tannahill CL, Mallon E, Mcglynn LM, Pyne S, Pyne NJ and Edwards J: O-56 High expression of sphingosine 1-phosphate receptors, S1P 1 and S1P 3, sphingosine kinase 1 and ERK-1/2 is associated with development of tamoxifen resistance in ER positive breast cancer patients. Eur J Cancer Suppl. 8:212010. View Article : Google Scholar
24	Yoshida Y, Nakada M, Harada T, Tanaka S, Furuta T, Hayashi Y, Kita D, Uchiyama N, Hayashi Y and Hamada J: The expression level of sphingosine-1-phosphate receptor type 1 is related to MIB-1 labeling index and predicts survival of glioblastoma patients. J Neurooncol. 98:41–47. 2010. View Article : Google Scholar
25	Kothapalli R, Kusmartseva I and Loughran TP: Characterization of a human sphingosine-1-phosphate receptor gene (S1P 5) and its differential expression in LGL leukemia. Biochim Biophys Acta. 1579:117–123. 2002. View Article : Google Scholar
26	Cattoretti G, Mandelbaum J, Lee N, Chaves AH, Mahler AM, Chadburn A, Dallafavera R, Pasqualucci L and Maclennan AJ: Targeted disruption of the S1P2 sphingosine 1-phosphate receptor gene leads to diffuse large B-cell lymphoma formation. Cancer Res. 69:8686–8692. 2009. View Article : Google Scholar : PubMed/NCBI
27	Flori M, Schmid CA, Sumrall ET, Tzankov A, Law CW, Robinson MD and Muller A: The hematopoietic oncoprotein FOXP1 promotes tumor cell survival in diffuse large B-cell lymphoma by repressing S1PR2 signaling. Blood. 127:1438–1448. 2016. View Article : Google Scholar
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
29	Kawamori T, Kaneshiro T, Okumura M, Maalouf S, Uflacker A, Bielawski J, Hannun YA and Obeid LM: Role for sphingosine kinase 1 in colon carcinogenesis. FASEB J. 23:405–414. 2009. View Article : Google Scholar : PubMed/NCBI
30	Bao M, Chen Z, Xu Y, Zhao Y, Zha R, Huang S, Liu L, Chen T, Li J, Tu H and He X: Sphingosine kinase 1 promotes tumour cell migration and invasion via the S1P/EDG1 axis in hepatocellular carcinoma. Liver Int. 32:331–338. 2012. View Article : Google Scholar
31	Terashita T, Kobayashi K, Nagano T, Kawa Y, Tamura D, Nakata K, Yamamoto M, Tachihara M, Kamiryo H and Nishimura Y: Administration of JTE013 abrogates experimental asthma by regulating proinflammatory cytokine production from bronchial epithelial cells. Respir Res. 17:1462016. View Article : Google Scholar : PubMed/NCBI
32	Zhang X, Wang W, Ji XY, Ritter JK and Li N: Knockout of sphingosine kinase 1 attenuates renal fibrosis in unilateral ureteral obstruction model. Am J Nephrol. 50:196–203. 2019. View Article : Google Scholar
33	Nuno DW and Lamping KG: Dietary fatty acid saturation modulates sphingosine-1-phosphate-mediated vascular function. J Diabetes Res. 2019:23542742019. View Article : Google Scholar : PubMed/NCBI
34	Salas A, Ponnusamy S, Senkal CE, Meyers-Needham M, Selvam SP, Saddoughi SA, Apohan E, Sentelle RD, Smith C, Gault CR, et al: Sphingosine kinase-1 and sphingosine 1-phosphate receptor 2 mediate Bcr-Abl1 stability and drug resistance by modulation of protein phosphatase 2A. Blood. 117:5941–5952. 2011. View Article : Google Scholar
35	Jozefczuk E, Nosalski R, Saju B, Crespo E, Szczepaniak P, Guzik TJ and Siedlinski M: Cardiovascular effects of pharmacological targeting of sphingosine kinase 1. Hypertension. 75:383–392. 2020. View Article : Google Scholar : PubMed/NCBI
36	Kunkel GT, Maceyka M, Milstien S and Spiegel S: Targeting the sphingosine-1-phosphate axis in cancer, inflammation and beyond. Nat Rev Drug Discov. 12:688–702. 2013. View Article : Google Scholar : PubMed/NCBI
37	Perla AS, Fratini L, Cardoso PS, de Farias CB, Da CJM and Roesler R: Fingolimod (FTY720) reduces viability and survival and increases histone H3 acetylation in medulloblastoma cells. Pediatr Hematol Oncol. 37:170–175. 2020. View Article : Google Scholar
38	Pyne S and Pyne NJ: New Perspectives on the Role of Sphingosine 1-Phosphate in Cancer. Sphingolipids in Disease. Handbook of Experimental Pharmacology. Gulbins E and Petrache I: 216. Springer; Vienna: pp. 55–71. 2013, View Article : Google Scholar
39	Beach JA, Aspuria PJ, Cheon DJ, Lawrenson K, Agadjanian H, Walsh CS, Karlan BY and Orsulic S: Sphingosine kinase 1 is required for TGF-beta mediated fibroblastto-myofibroblast differentiation in ovarian cancer. Oncotarget. 7:4167–4182. 2016. View Article : Google Scholar
40	Shida D, Takabe K, Kapitonov D, Milstien S and Spiegel S: Targeting SphK1 as a new strategy against cancer. Curr Drug Targets. 9:662–673. 2008. View Article : Google Scholar : PubMed/NCBI
41	Chen MH, Yen CC, Cheng CT, Wu RC, Huang SC, Yu CS, Chung YH, Liu CY, Chang PM, Chao Y, et al: Identification of SPHK1 as a therapeutic target and marker of poor prognosis in cholangiocarcinoma. Oncotarget. 6:23594–23608. 2015. View Article : Google Scholar
42	Patmanathan SN, Johnson SP, Lai SL, Bernam SP, Lopes V, Wei W, Ibrahim MH, Torta F, Narayanaswamy P and Wenk MR: Aberrant expression of the S1P regulating enzymes, SPHK1 and SGPL1, contributes to a migratory phenotype in OSCC mediated through S1PR2. Sci Rep. 6:256502016. View Article : Google Scholar : PubMed/NCBI
43	Ruckhäberle E, Rody A, Engels K, Gaetje R, von Minckwitz G, Schiffmann S, Grösch S, Geisslinger G, Holtrich U, Karn T and Kaufmann M: Microarray analysis of altered sphingolipid metabolism reveals prognostic significance of sphingosine kinase 1 in breast cancer. Breast Cancer Res Treat. 112:41–52. 2008. View Article : Google Scholar
44	Furuya H, Tamashiro PM, Shimizu Y, Iino K, Peres R, Chen R, Sun Y, Hannun YA, Obeid LM and Kawamori T: Sphingosine Kinase 1 expression in peritoneal macrophages is required for colon carcinogenesis. Carcinogenesis. 38:1218–1227. 2017. View Article : Google Scholar : PubMed/NCBI
45	Furuya H, Shimizu Y, Tamashiro PM, Iino K, Bielawski J, Chan O, Pagano I and Kawamori T: Sphingosine kinase 1 expression enhances colon tumor growth. J Transl Med. 15:1202017. View Article : Google Scholar : PubMed/NCBI
46	Weigert A, Olesch C and Brune B: Sphingosine-1-Phosphate and macrophage biology-how the sphinx tames the big eater. Front Immunol. 10:17062019. View Article : Google Scholar
47	Andrieu G, Ledoux A, Branka S, Bocquet M, Gilhodes J, Walzer T, Kasahara K, Inagaki M, Sabbadini RA, Cuvillier O and Hatzoglou A: Sphingosine 1-phosphate signaling through its receptor S1P5 promotes chromosome segregation and mitotic progression. Sci Signal. 10:eaah40072017. View Article : Google Scholar
48	Suh JH and Saba JD: Sphingosine-1-phosphate in inflammatory bowel disease and colitis-associated colon cancer: The fat's in the fire. Transl Cancer Res. 4:469–483. 2015.PubMed/NCBI
49	Wang C, Mao J, Redfield S, Mo Y, Lage JM and Zhou X: Systemic distribution, subcellular localization and differential expression of sphingosine-1-phosphate receptors in benign and malignant human tissues. Exp Mol Pathol. 97:259–265. 2014. View Article : Google Scholar
50	Herr DR, Reolo MJ, Peh YX, Wang W, Lee CW, Rivera R, Paterson IC and Chun J: Sphingosine 1-phosphate receptor 2 (S1P2) attenuates reactive oxygen species formation and inhibits cell death: Implications for otoprotective therapy. Sci Rep. 6:245412016. View Article : Google Scholar : PubMed/NCBI
51	Brinkmann V, Billich A, Baumruker T, Heining P, Schmouder R, Francis G, Aradhye S and Burtin P: Fingolimod (FTY720): Discovery and development of an oral drug to treat multiple sclerosis. Nat Rev Drug Discov. 9:883–897. 2010. View Article : Google Scholar : PubMed/NCBI
52	Barthelemy C, Barry AO, Twyffels L and Andre B: FTY720-induced endocytosis of yeast and human amino acid transporters is preceded by reduction of their inherent activity and TORC1 inhibition. Sci Rep. 7:138162017. View Article : Google Scholar : PubMed/NCBI