Activation of the SphK1/ERK/p‑ERK pathway promotes autophagy in colon cancer cells

Xu,Chunyan; Zhang,Wenlu; Liu,Shiquan; Wu,Wenhong; Qin,Mengbin; Huang,Jiean

doi:10.3892/ol.2018.8588

Activation of the SphK1/ERK/p‑ERK pathway promotes autophagy in colon cancer cells

Authors:
- Chunyan Xu
- Wenlu Zhang
- Shiquan Liu
- Wenhong Wu
- Mengbin Qin
- Jiean Huang
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Affiliations: Department of Gastroenterology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530007, P.R. China, Department of Respiratory Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: April 26, 2018 https://doi.org/10.3892/ol.2018.8588
Pages: 9719-9724
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Sphingosine kinase 1 (SphK1) is a master kinase that catalyzes the synthesis of sphingosine 1 phosphate and participates in the regulation of cell proliferation and autophagy. The present study aimed to assess the effects of the activation of the SphK1/extracellular signal‑regulated kinase (ERK)/phosphorylated (p‑)ERK pathway in the regulation of autophagy in colon cancer (HT‑29) cells. Inverted fluorescence microscopy was used to detect the expression of green fluorescent protein (GFP) in the SphK1‑overexpressing HT‑29 cells [SphK1(+)‑HT‑29] and the negative control HT‑29 cells (NC‑HT‑29). Western blotting was used to detect the protein expression levels of SphK1, ERK1/2, p‑ERK1/2, as well as those of the autophagy‑associated markers LC3A, ATG5, and ULK1. Protein localization and expression of the LC3A antibody were detected by immunofluorescence. The results demonstrated that GFP was similarly expressed in SphK1(+)‑HT‑29 and NC‑HT‑29 cells. However, significantly increased SphK1 mRNA and protein expression levels were detected in SphK1(+)‑HT‑29 cells compared with in NC‑HT‑29 cells, which resulted in upregulated ERK/p‑ERK. Furthermore, the protein expression levels of the three autophagy‑associated markers increased. LC3A protein was localized in the cytoplasm of SphK1(+)‑HT‑29 cells, indicating autophagy. In summary, the findings of the present study suggested that activation of the SphK1/ERK/p‑ERK pathway promotes autophagy in colon cancer HT‑29 cells.

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1	Nagappan A, Lee WS, Yun JW, Lu JN, Chang SH, Jeong JH, Kim GS, Jung JM and Hong SC: Tetraarsenic hexoxide induces G2/M arrest, apoptosis, and autophagy via PI3K/Akt suppression and p38 MAPK activation in SW620 human colon cancer cells. PLoS One. 12:e01745912017. View Article : Google Scholar : PubMed/NCBI
2	Izumi M and Nakamura S: Chloroplast protein turnover: The influence of extraplastidic processes, including autophagy. Int J Mol Sci. 19:E8282018. View Article : Google Scholar : PubMed/NCBI
3	Lampada A, O'Prey J, Szabadkai G, Ryan KM, Hochhauser D and Salomoni P: mTORC1-independent autophagy regulates receptor tyrosine kinase phosphorylation in colorectal cancer cells via an mTORC2-mediated mechanism. Cell Death Differ. 24:1045–1062. 2017. View Article : Google Scholar : PubMed/NCBI
4	He M, Luo M, Liu Q, Chen J, Li K, Zheng M, Weng Y, Ouyang L and Liu A: Combination treatment with fasudil and clioquinol produces synergistic anti-tumor effects in U87 glioblastoma cells by activating apoptosis and autophagy. J Neurooncol. 127:261–270. 2016. View Article : Google Scholar : PubMed/NCBI
5	Cheng YC, Hueng DY, Huang HY, Chen JY and Chen Y: Magnolol and honokiol exert a synergistic anti-tumor effect through autophagy and apoptosis in human glioblastomas. Oncotarget. 7:29116–29130. 2016.PubMed/NCBI
6	Zhang L, Yu Y, Xia X, Ma Y, Chen XW, Ni ZH and Wang H: Transcription factor E2-2 inhibits the proliferation of endothelial progenitor cells by suppressing autophagy. Int J Mol Med. 37:1254–1262. 2016. View Article : Google Scholar : PubMed/NCBI
7	Samardzija G, Stevovic TK, Djuricic S, Djokic D, Djurisic M, Ciric D, Martinovic T, Bumbasirevic V and Vujic D: Aggressive human neuroblastomas show a massive increase in the numbers of autophagic vacuoles and damaged mitochondria. Ultrastruct Pathol. 40:240–248. 2016. View Article : Google Scholar : PubMed/NCBI
8	Long J, Xie Y, Yin J, Lu W and Fang S: SphK1 promotes tumor cell migration and invasion in colorectal cancer. Tumour Biol. 37:6831–6836. 2016. View Article : Google Scholar : PubMed/NCBI
9	Almejún MB, Borge M, Colado A, Elías EE, Podaza E, Risnik D, De Brasi CD, Stanganelli C, Slavutsky I, Cabrejo M, et al: Sphingosine kinase 1 participates in the activation, proliferation and survival of chronic lymphocytic leukemia cells. Haematologica. 102:e257–e260. 2017. View Article : Google Scholar : PubMed/NCBI
10	Manchon Moruno JF, Uzor NE, Finkbeiner S and Tsvetkov AS: SPHK1/sphingosine kinase 1-mediated autophagy differs between neurons and SH-SY5Y neuroblastoma cells. Autophagy. 12:1418–1424. 2016. View Article : Google Scholar : PubMed/NCBI
11	Lavieu G, Scarlatti F, Sala G, Carpentier S, Levade T, Ghidoni R, Botti J and Codogno P: Regulation of autophagy by sphingosine kinase 1 and its role in cell survival during nutrient starvation. J Biol Chem. 281:8518–8527. 2006. View Article : Google Scholar : PubMed/NCBI
12	Ren H, Guo H, Thakur A, Zhang S, Wang T, Liang Y, Shi P, Gao L, Liu F, Feng J, et al: Blockade efficacy of MEK/ERK-dependent autophagy enhances PI3K/Akt inhibitor NVP-BKM120's therapeutic effectiveness in lung cancer cells. Oncotarget. 7:67277–67287. 2016. View Article : Google Scholar : PubMed/NCBI
13	Zhao Y, Fan D, Zheng ZP, Li ET, Chen F, Cheng KW and Wang M: 8-C-(E-phenylethenyl)quercetin from onion/beef soup induces autophagic cell death in colon cancer cells through ERK activation. Mol Nutr Food Res. 61:2017. View Article : Google Scholar
14	Liu SQ, Huang JA, Qin MB, Su YJ, Lai MY, Jiang HX and Tang GD: Sphingosine kinase 1 enhances colon cancer cell proliferation and invasion by upregulating the production of MMP-2/9 and uPA via MAPK pathways. Int J Colorectal Dis. 27:1569–1578. 2012. View Article : Google Scholar : PubMed/NCBI
15	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
16	Xu CY, Liu SQ, Qin MB, Zhuge CF, Qin L, Qin N, Lai MY and Huang JA: SphK1 modulates cell migration and EMT-related marker expression by regulating the expression of p-FAK in colorectal cancer cells. Int J Mol Med. 39:1277–1284. 2017. View Article : Google Scholar : PubMed/NCBI
17	Nihira K, Miki Y, Iida S, Narumi S, Ono K, Iwabuchi E, Ise K, Mori K, Saito M, Ebina M, et al: An activation of LC3A-mediated autophagy contributes to de novo and acquired resistance to EGFR tyrosine kinase inhibitors in lung adenocarcinoma. J Pathol. 234:277–288. 2014.PubMed/NCBI
18	Miyamoto M, Takano M, Aoyama T, Soyama H, Yoshikawa T, Tsuda H and Furuya K: Inhibition of autophagy protein LC3A as a therapeutic target in ovarian clear cell carcinomas. J Gynecol Oncol. 28:e332017. View Article : Google Scholar : PubMed/NCBI
19	Giatromanolaki A, Koukourakis MI, Pouliliou S, Gatter KC, Pezzella F, Harris AL and Sivridis E: Overexpression of LC3A autophagy protein in follicular and diffuse large B-cell lymphomas. Hematol Oncol Stem Cell Ther. 6:20–25. 2013. View Article : Google Scholar : PubMed/NCBI
20	Martinez-Lopez N, Athonvarangkul D, Mishall P, Sahu S and Singh R: Autophagy proteins regulate ERK phosphorylation. Nat Commun. 4:27992013. View Article : Google Scholar : PubMed/NCBI
21	Tanida I, Ueno T and Kominami E: LC3 and autophagy. Methods Mol Biol. 445:77–88. 2008. View Article : Google Scholar : PubMed/NCBI
22	Mowers EE, Sharifi MN and Macleod KF: Autophagy in cancer metastasis. Oncogene. 36:1619–1630. 2017. View Article : Google Scholar : PubMed/NCBI
23	Yu Y, Hou L, Song H, Xu P, Sun Y and Wu K: Akt/AMPK/mTOR pathway was involved in the autophagy induced by vitamin E succinate in human gastric cancer SGC-7901 cells. Mol Cell Biochem. 424:173–183. 2017. View Article : Google Scholar : PubMed/NCBI
24	Wang R, Shen Z, Yang L, Yin M, Zheng W, Wu B, Liu T and Song H: Protective effects of heme oxygenase-1-transduced bone marrow-derived mesenchymal stem cells on reducedsize liver transplantation: Role of autophagy regulated by the ERK/mTOR signaling pathway. Int J Mol Med. 40:1537–1548. 2017. View Article : Google Scholar : PubMed/NCBI
25	Nazio F and Cecconi F: Autophagy up and down by outsmarting the incredible ULK. Autophagy. 13:967–968. 2017. View Article : Google Scholar : PubMed/NCBI
26	Bao Y, Li K, Guo Y, Wang Q, Li Z, Yang Y, Chen Z, Wang J, Zhao W, Zhang H, et al: Tumor suppressor PRSS8 targets Sphk1/S1P/Stat3/Akt signaling in colorectal cancer. Oncotarget. 7:26780–26792. 2016. View Article : Google Scholar : PubMed/NCBI
27	González-Fernández B, Sánchez DI, Crespo I, San-Miguel B, Álvarez M, Tuñón MJ and González-Gallego J: Inhibition of the SphK1/S1P signaling pathway by melatonin in mice with liver fibrosis and human hepatic stellate cells. Biofactors. 43:272–282. 2017. View Article : Google Scholar : PubMed/NCBI
28	Gao Z, Wang H, Xiao FJ, Shi XF, Zhang YK, Xu QQ, Zhang XY, Ha XQ and Wang LS: SIRT1 mediates Sphk1/S1P-induced proliferation and migration of endothelial cells. Int J Biochem Cell Biol. 74:152–160. 2016. View Article : Google Scholar : PubMed/NCBI
29	Buchegger K, Silva R, López J, Ili C, Araya JC, Leal P, Brebi P, Riquelme I and Roa JC: The ERK/MAPK pathway is overexpressed and activated in gallbladder cancer. Pathol Res Pract. 213:476–482. 2017. View Article : Google Scholar : PubMed/NCBI
30	Maeda Y, Oku M and Sakai Y: Autophagy-independent function of Atg8 in lipid droplet dynamics in yeast. J Biochem. 161:339–348. 2017.PubMed/NCBI
31	Li L, Chen H, Gao Y, Wang YW, Zhang GQ, Pan SH, Ji L, Kong R, Wang G, Jia YH, et al: Long noncoding RNA MALAT1 promotes aggressive pancreatic cancer proliferation and metastasis via the stimulation of autophagy. Mol Cancer Ther. 15:2232–2243. 2016. View Article : Google Scholar : PubMed/NCBI
32	Kwon YT and Ciechanover A: The ubiquitin code in the ubiquitin-proteasome system and autophagy. Trends Biochem Sci. 42:873–886. 2017. View Article : Google Scholar : PubMed/NCBI