C16‑ceramide and sphingosine 1‑phosphate/S1PR2 have opposite effects on cell growth through mTOR signaling pathway regulation

Kim,Min Hee; Park,Joo‑Won; Lee,Eun‑Ji; Kim,Shin; Shin,Sun‑Hye; Ahn,Jung‑Hyuck; Jung,Yunjae; Park,Inkeun; Park,Woo‑Jae

doi:10.3892/or.2018.6689

C16‑ceramide and sphingosine 1‑phosphate/S1PR2 have opposite effects on cell growth through mTOR signaling pathway regulation

Authors:
- Min Hee Kim
- Joo‑Won Park
- Eun‑Ji Lee
- Shin Kim
- Sun‑Hye Shin
- Jung‑Hyuck Ahn
- Yunjae Jung
- Inkeun Park
- Woo‑Jae Park
View Affiliations

Affiliations: Department of Biochemistry, College of Medicine, Gachon University, Incheon 21999, Republic of Korea, Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07985, Republic of Korea, Department of Biochemistry, College of Medicine, Gachon University, Incheon 21999, Republic of Korea, Department of Immunology, School of Medicine, Keimyung University, Daegu 42601, Republic of Korea, Department of Microbiology, College of Medicine, Gachon University, Incheon 21999, Division of Medical Oncology, Department of Internal Medicine, Gachon University Gil Medical Center, Incheon 21565, Republic of Korea , Division of Medical Oncology, Department of Internal Medicine, Gachon University Gil Medical Center, Incheon 21565, Republic of Korea
Published online on: September 7, 2018 https://doi.org/10.3892/or.2018.6689
Pages: 2977-2987

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

Recently, sphingolipid derivatives, such as ceramide and sphingosine‑1‑phosphate (S1P), have emerged as key modulators in apoptotic cell death and cell proliferation. This study aimed to clarify the underlying signaling pathways of ceramide and S1P involved in breast cancer cell proliferation. Ceramide acyl chain length is determined by six mammalian ceramide synthases (CerS). We overexpressed CerS1 to 6 in MCF‑7 cells to examine whether ceramide signaling propagation varies as a function of acyl chain length. Among the six CerS, only CerS6 overexpression reduced phosphorylation of Akt, S6 kinase (S6K), and extracellular signal‑regulated kinases (ERK) as shown by western blotting. In addition, CerS6 overexpression reduced MCF‑7 cell proliferation. This effect was partially reversed by co‑treatment with MHY1485, an activator of mammalian target of rapamycin (mTOR), demonstrating an important role for the mTOR pathway in the CerS6‑mediated decrease in MCF‑7 cell proliferation. ERK inhibition, but not Akt inhibition, along with mTOR inhibition synergistically reduced MCF‑7 cell proliferation as measured by MTT assay. Notably, the expression of CerS6 and S1P receptor 2 (S1PR2), or CerS6 and sphingosine kinase 1 (SphK1), were negatively correlated according to the invasive breast carcinoma patient cohort in The Cancer Genome Atlas database. In addition, both SphK1 overexpression and S1P addition increased mTOR phosphorylation as shown by ELISA, while S1PR2 inhibition had the inverse effect. These data suggest that CerS6 and SphK1 regulate mTOR signaling in breast cancer cell proliferation. Moreover, mTOR activity can be regulated by the balance between S1P and C16‑ceramide, which is generated by CerS6.

View Figures

View References

1	Li D, Bi FF, Chen NN, Cao JM, Sun WP, Zhou YM, Li CY and Yang Q: A novel crosstalk between BRCA1 and poly (ADP-ribose) polymerase 1 in breast cancer. Cell Cycle. 13:3442–3449. 2014. View Article : Google Scholar : PubMed/NCBI
2	Saini KS, Loi S, de Azambuja E, Metzger-Filho O, Saini ML, Ignatiadis M, Dancey JE and Piccart-Gebhart MJ: Targeting the PI3K/AKT/mTOR and Raf/MEK/ERK pathways in the treatment of breast cancer. Cancer Treat Rev. 39:935–946. 2013. View Article : Google Scholar : PubMed/NCBI
3	Weinstein-Oppenheimer CR, Burrows C, Steelman LS and McCubrey JA: The effects of beta-estradiol on Raf activity, cell cycle progression and growth factor synthesis in the MCF-7 breast cancer cell line. Cancer Biol Ther. 1:256–262. 2002. View Article : Google Scholar : PubMed/NCBI
4	Hurvitz SA, Kalous O, Conklin D, Desai AJ, Dering J, Anderson L, O'Brien NA, Kolarova T, Finn RS, Linnartz R, et al: In vitro activity of the mTOR inhibitor everolimus, in a large panel of breast cancer cell lines and analysis for predictors of response. Breast Cancer Res Treat. 149:669–680. 2015. View Article : Google Scholar : PubMed/NCBI
5	Liu H, Hu C, Wu X and Li Z: Equol elicits estrogenic activities via PI3K/akt pathway in the estrogen receptor-positive MCF-7 cells. Mol Cell Toxicol. 10:285–291. 2014. View Article : Google Scholar
6	Paplomata E and O'Regan R: The PI3K/AKT/mTOR pathway in breast cancer: Targets, trials and biomarkers. Ther Adv Med Oncol. 6:154–166. 2014. View Article : Google Scholar : PubMed/NCBI
7	Bjornsti MA and Houghton PJ: The TOR pathway: A target for cancer therapy. Nat Rev Cancer. 4:335–348. 2004. View Article : Google Scholar : PubMed/NCBI
8	Newton J, Lima S, Maceyka M and Spiegel S: Revisiting the sphingolipid rheostat: Evolving concepts in cancer therapy. Exp Cell Res. 333:195–200. 2015. View Article : Google Scholar : PubMed/NCBI
9	Ogretmen B: Sphingolipid metabolism in cancer signalling and therapy. Nat Rev Cancer. 18:33–50. 2018. View Article : Google Scholar : PubMed/NCBI
10	Park WJ and Park JW: The effect of altered sphingolipid acyl chain length on various disease models. Biol Chem. 396:693–705. 2015. View Article : Google Scholar : PubMed/NCBI
11	Park JW, Park WJ and Futerman AH: Ceramide synthases as potential targets for therapeutic intervention in human diseases. Biochim Biophys Acta. 1841:671–681. 2014. View Article : Google Scholar : PubMed/NCBI
12	Karahatay S, Thomas K, Koybasi S, Senkal CE, Elojeimy S, Liu X, Bielawski J, Day TA, Gillespie MB, Sinha D, et al: Clinical relevance of ceramide metabolism in the pathogenesis of human head and neck squamous cell carcinoma (HNSCC): Attenuation of C₁₈-ceramide in HNSCC tumors correlates with lymphovascular invasion and nodal metastasis. Cancer Lett. 256:101–111. 2007. View Article : Google Scholar : PubMed/NCBI
13	Mesicek J, Lee H, Feldman T, Jiang X, Skobeleva A, Berdyshev EV, Haimovitz-Friedman A, Fuks Z and Kolesnick R: Ceramide synthases 2, 5, and 6 confer distinct roles in radiation-induced apoptosis in HeLa cells. Cell Signal. 22:1300–1307. 2010. View Article : Google Scholar : PubMed/NCBI
14	Hartmann D, Lucks J, Fuchs S, Schiffmann S, Schreiber Y, Ferreirós N, Merkens J, Marschalek R, Geisslinger G and Grösch S: Long chain ceramides and very long chain ceramides have opposite effects on human breast and colon cancer cell growth. Int J Biochem Cell Biol. 44:620–628. 2012. View Article : Google Scholar : PubMed/NCBI
15	Zigdon H, Kogot-Levin A, Park JW, Goldschmidt R, Kelly S, Merrill AH Jr, Scherz A, Pewzner-Jung Y, Saada A and Futerman AH: Ablation of ceramide synthase 2 causes chronic oxidative stress due to disruption of the mitochondrial respiratory chain. J Biol Chem. 288:4947–4956. 2013. View Article : Google Scholar : PubMed/NCBI
16	Raichur S, Wang ST, Chan PW, Li Y, Ching J, Chaurasia B, Dogra S, Öhman MK, Takeda K, Sugii S, et al: CerS2 haploinsufficiency inhibits β-oxidation and confers susceptibility to diet-induced steatohepatitis and insulin resistance. Cell Metab. 20:9192014. View Article : Google Scholar : PubMed/NCBI
17	Park WJ, Park JW, Merrill AH, Storch J, Pewzner-Jung Y and Futerman AH: Hepatic fatty acid uptake is regulated by the sphingolipid acyl chain length. Biochim Biophys Acta. 1841:1754–1766. 2014. View Article : Google Scholar : PubMed/NCBI
18	Maceyka M, Harikumar KB, Milstien S and Spiegel S: Sphingosine-1-phosphate signaling and its role in disease. Trends Cell Biol. 22:50–60. 2012. View Article : Google Scholar : PubMed/NCBI
19	Strub GM, Maceyka M, Hait NC, Milstien S and Spiegel S: Extracellular and intracellular actions of sphingosine-1-phosphate. Adv Exp Med Biol. 688:141–155. 2010. View Article : Google Scholar : PubMed/NCBI
20	Nagahashi M, Tsuchida J, Moro K, Hasegawa M, Tatsuda K, Woelfel IA, Takabe K and Wakai T: High levels of sphingolipids in human breast cancer. J Surg Res. 204:435–444. 2016. View Article : Google Scholar : PubMed/NCBI
21	Erez-Roman R, Pienik R and Futerman AH: Increased ceramide synthase 2 and 6 mRNA levels in breast cancer tissues and correlation with sphingosine kinase expression. Biochem Biophys Res Commun. 391:219–223. 2010. View Article : Google Scholar : PubMed/NCBI
22	Fan S, Niu Y, Tan N, Wu Z, Wang Y, You H, Ke R, Song J, Shen Q, Wang W, et al: LASS2 enhances chemosensitivity of breast cancer by counteracting acidic tumor microenvironment through inhibiting activity of V-ATPase proton pump. Oncogene. 32:1682–1690. 2013. View Article : Google Scholar : PubMed/NCBI
23	Fan SH, Wang YY, Lu J, Zheng YL, Wu DM, Zhang ZF, Shan Q, Hu B, Li MQ and Cheng W: CERS2 suppresses tumor cell invasion and is associated with decreased V-ATPase and MMP-2/MMP-9 activities in breast cancer. J Cell Biochem. 116:502–513. 2015. View Article : Google Scholar : PubMed/NCBI
24	Fekry B, Esmaeilniakooshkghazi A, Krupenko SA and Krupenko NI: Ceramide synthase 6 is a novel target of methotrexate mediating its antiproliferative effect in a p53-dependent manner. PLoS One. 11:e01466182016. View Article : Google Scholar : PubMed/NCBI
25	Maeng HJ, Song JH, Kim GT, Song YJ, Lee K, Kim JY and Park TS: Celecoxib-mediated activation of endoplasmic reticulum stress induces de novo ceramide biosynthesis and apoptosis in hepatoma HepG2 cells mobilization. BMB Rep. 50:144–149. 2017. View Article : Google Scholar : PubMed/NCBI
26	Denard B, Lee C and Ye J: Doxorubicin blocks proliferation of cancer cells through proteolytic activation of CREB3L1. Elife. 1:e000902012. View Article : Google Scholar : PubMed/NCBI
27	Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
28	Cerami E, Gao J, Dogrusoz U, Gross BE, Sumer SO, Aksoy BA, Jacobsen A, Byrne CJ, Heuer ML, Larsson E, et al: The cBio cancer genomics portal: An open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2:401–404. 2012. View Article : Google Scholar : PubMed/NCBI
29	Merrill AH, van Echten G, Wang E and Sandhoff K: Fumonisin B1 inhibits sphingosine (sphinganine) N-acyltransferase and de novo sphingolipid biosynthesis in cultured neurons in situ. J Biol Chem. 268:27299–27306. 1993.PubMed/NCBI
30	Cancer Genome Atlas Network, . Koboldt DC, Fulton RS, McLellan MD, Schmidt H, Kalicki-Veizer J, McMichael JF, Fulton LL, Dooling DJ, Ding L, Mardis ER, et al: Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
31	Woodcock J: Sphingosine and ceramide signalling in apoptosis. IUBMB Life. 58:462–466. 2006. View Article : Google Scholar : PubMed/NCBI
32	Shayman JA: Sphingolipids. Kidney Int. 58:11–26. 2000. View Article : Google Scholar : PubMed/NCBI
33	Toss A and Cristofanilli M: Molecular characterization and targeted therapeutic approaches in breast cancer. Breast Cancer Res. 17:602015. View Article : Google Scholar : PubMed/NCBI
34	Mendoza MC, Er EE and Blenis J: The Ras-ERK and PI3K-mTOR pathways: Cross-talk and compensation. Trends Biochem Sci. 36:320–328. 2011. View Article : Google Scholar : PubMed/NCBI
35	Serra V, Scaltriti M, Prudkin L, Eichhorn PJ, Ibrahim YH, Chandarlapaty S, Markman B, Rodriguez O, Guzman M, Rodriguez S, et al: PI3K inhibition results in enhanced HER signaling and acquired ERK dependency in HER2-overexpressing breast cancer. Oncogene. 30:2547–2557. 2011. View Article : Google Scholar : PubMed/NCBI
36	Baselga J, Campone M, Piccart M, Burris HA III, Rugo HS, Sahmoud T, Noguchi S, Gnant M, Pritchard KI, Lebrun F, et al: Everolimus in postmenopausal hormone-receptor-positive advanced breast cancer. N Engl J Med. 366:520–529. 2012. View Article : Google Scholar : PubMed/NCBI
37	Piccart M, Hortobagyi GN, Campone M, Pritchard KI, Lebrun F, Ito Y, Noguchi S, Perez A, Rugo HS, Deleu I, et al: Everolimus plus exemestane for hormone-receptor-positive, human epidermal growth factor receptor-2-negative advanced breast cancer: Overall survival results from BOLERO-2†. Ann Oncol. 25:2357–2362. 2014. View Article : Google Scholar : PubMed/NCBI
38	Min J, Mesika A, Sivaguru M, Van Veldhoven PP, Alexander H, Futerman AH and Alexander S: (Dihydro)ceramide synthase 1 regulated sensitivity to cisplatin is associated with the activation of p38 mitogen-activated protein kinase and is abrogated by sphingosine kinase 1. Mol Cancer Res. 5:801–812. 2007. View Article : Google Scholar : PubMed/NCBI
39	Blaho VA and Hla T: An update on the biology of sphingosine 1-phosphate receptors. J Lipid Res. 55:1596–1608. 2014. View Article : Google Scholar : PubMed/NCBI
40	Schiffmann S, Sandner J, Birod K, Wobst I, Angioni C, Ruckhäberle E, Kaufmann M, Ackermann H, Lötsch J, Schmidt H, et al: Ceramide synthases and ceramide levels are increased in breast cancer tissue. Carcinogenesis. 30:745–752. 2009. View Article : Google Scholar : PubMed/NCBI
41	Gupta V, Bhinge KN, Hosain SB, Xiong K, Gu X, Shi R, Ho MY, Khoo KH, Li SC, Li YT, et al: Ceramide glycosylation by glucosylceramide synthase selectively maintains the properties of breast cancer stem cells. J Biol Chem. 287:37195–37205. 2012. View Article : Google Scholar : PubMed/NCBI
42	Novgorodov SA, Chudakova DA, Wheeler BW, Bielawski J, Kindy MS, Obeid LM and Gudz TI: Developmentally regulated ceramide synthase 6 increases mitochondrial Ca²⁺ loading capacity and promotes apoptosis. J Biol Chem. 286:4644–4658. 2011. View Article : Google Scholar : PubMed/NCBI
43	Yu J, Novgorodov SA, Chudakova D, Zhu H, Bielawska A, Bielawski J, Obeid LM, Kindy MS and Gudz TI: JNK3 signaling pathway activates ceramide synthase leading to mitochondrial dysfunction. J Biol Chem. 282:25940–25949. 2007. View Article : Google Scholar : PubMed/NCBI