Exosomes from LNCaP cells promote osteoblast activity through miR‑375 transfer

Li,Su‑Liang; An,Na; Liu,Bing; Wang,Sheng‑Yu; Wang,Jian‑Jun; Ye,Yun

doi:10.3892/ol.2019.10110

Exosomes from LNCaP cells promote osteoblast activity through miR‑375 transfer

Authors:
- Su‑Liang Li
- Na An
- Bing Liu
- Sheng‑Yu Wang
- Jian‑Jun Wang
- Yun Ye
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Affiliations: Department of Clinical Laboratory, The First Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710077, P.R. China, Department of Laboratory Medicine, Shaanxi Jiaotong Hospital, Xi'an, Shaanxi 710068, P.R. China, Department of Pathology, The First Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710077, P.R. China, Intensive Care Unit, The First Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710077, P.R. China
Published online on: March 5, 2019 https://doi.org/10.3892/ol.2019.10110
Pages: 4463-4473
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Previous studies have revealed that exosomes influence tumour metastasis, diagnosis and treatment. In addition, exosomal microRNAs (miRNAs/miRs) are closely associated with the metastatic microenvironment; however, the regulatory role of exosomal miRNAs from prostate cancer cells on bone metastasis remains poorly understood. In the present study, a series of experiments were performed to determine whether exosomal miR‑375 from LNCaP cells promote osteoblast activity. Exosomes were isolated and purified by ultracentrifugation, total RNA from cells and total miRNA from exosomes were then extracted, and miR‑375 levels were detected by reverse transcription‑quantitative polymerase chain reaction. Exosome libraries from LNCaP and RWPE‑1 cells were sequenced and selected using an Illumina HiSeq™ 2500 system. The effects of exosomes on osteoblasts were determined and osteoblast activity was evaluated by measuring the activity of alkaline phosphatase, the extent of extracellular matrix mineralisation and the expression of osteoblast activity‑associated marker genes. Morphological observations, particle size analysis and molecular phenotyping confirmed that cell supernatants contained exosomes. Differential expression analysis confirmed high miR‑375 expression levels in LNCaP cell‑derived exosomes. The ability of exosomes to enter osteoblasts and increase their levels of miR‑375 was further analysed. The results demonstrated that exosomal miR‑375 significantly promoted osteoblast activity. In conclusion, the present study may lead to further investigation of the function role of exosomal miR‑375 in the activation and differentiation of osteoblasts in PCa.

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1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
2	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar : PubMed/NCBI
3	Suzman DL, Boikos SA and Carducci MA: Bone-targeting agents in prostate cancer. Cancer Metastasis Rev. 33:619–628. 2014. View Article : Google Scholar : PubMed/NCBI
4	Larson SR, Zhang X, Dumpit R, Coleman I, Lakely B, Roudier M, Higano CS, True LD, Lange PH, Montgomery B, et al: Characterization of osteoblastic and osteolytic proteins in prostate cancer bone metastases. Prostate. 73:932–940. 2013. View Article : Google Scholar : PubMed/NCBI
5	Rajpar S and Fizazi K: Bone targeted therapies in metastatic castration-resistant prostate cancer. Cancer J. 19:66–70. 2013. View Article : Google Scholar : PubMed/NCBI
6	Johnstone RM, Adam M, Hammond JR, Orr L and Turbide C: Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J Biol Chem. 262:9412–9420. 1987.PubMed/NCBI
7	Denzer K, Kleijmeer MJ, Heijnen HF, Stoorvogel W and Geuze HJ: Exosome: From internal vesicle of the multivesicular body to intercellular signaling device. J Cell Sci. 113:3365–3374. 2000.PubMed/NCBI
8	Gusachenko ON, Zenkova MA and Vlassov VV: Nucleic acids in exosomes: Disease markers and intercellular communication molecules. Biochemistry (Mosc). 78:1–7. 2013. View Article : Google Scholar : PubMed/NCBI
9	Soung YH, Ford S, Zhang V and Chung J: Exosomes in cancer diagnostics. Cancers (Basel). 9:82017. View Article : Google Scholar
10	Zhou Y, Xia L, Lin J, Wang H, Oyang L, Tan S, Tian Y, Su M, Wang H, Cao D and Liao Q: Exosomes in nasopharyngeal carcinoma. J Cancer. 9:767–777. 2018. View Article : Google Scholar : PubMed/NCBI
11	Ruivo CF, Adem B, Silva M and Melo SA: The Biology of Cancer Exosomes: Insights and New Perspectives. Cancer Res. 77:6480–6488. 2017. View Article : Google Scholar : PubMed/NCBI
12	Zhou M, Chen J, Zhou L, Chen W, Ding G and Cao L: Pancreatic cancer derived exosomes regulate the expression of TLR4 in dendritic cells via miR-203. Cell Immunol. 292:65–69. 2014. View Article : Google Scholar : PubMed/NCBI
13	Stevanato L, Thanabalasundaram L, Vysokov N and Sinden JD: Investigation of Content, Stoichiometry and Transfer of miRNA from Human Neural Stem CellLine Derived Exosomes. PLoS One. 11:e01463532016. View Article : Google Scholar : PubMed/NCBI
14	Greening DW, Gopal SK, Xu R, Simpson RJ and Chen W: Exosomes and their roles in immune regulation and cancer. Semin Cell Dev Biol. 40:72–81. 2015. View Article : Google Scholar : PubMed/NCBI
15	Hashimoto K, Ochi H, Sunamura S, Kosaka N, Mabuchi Y, Fukuda T, Yao K, Kanda H, Ae K, Okawa A, et al: Cancer-secreted hsa-miR-940 induces an osteoblastic phenotype in the bone metastatic microenvironment via targeting ARHGAP1 and FAM134A. Proc Natl Acad Sci USA. 115:2204–2209. 2018. View Article : Google Scholar : PubMed/NCBI
16	Corcoran C, Rani S and O'Driscoll L: miR-34a is an intracellular and exosomal predictive biomarker for response to docetaxel with clinical relevance to prostate cancer progression. Prostate. 74:1320–1334. 2014. View Article : Google Scholar : PubMed/NCBI
17	Ye Y, Li SL, Ma YY, Diao YJ, Yang L, Su MQ, Li Z, Ji Y, Wang J, Lei L, et al: Exosomal miR-141-3p regulates osteoblast activity to promote the osteoblastic metastasis of prostate cancer. Oncotarget. 8:94834–94849. 2017.PubMed/NCBI
18	Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur BK, Becker A, Hoshino A, Mark MT, Molina H, et al: Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Cell Bio. 17:816–826. 2015. View Article : Google Scholar
19	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. 25:402–8. 2001. View Article : Google Scholar : PubMed/NCBI
20	Möller A, House CM, Wong CS, Scanlon DB, Liu MC, Ronai Z and Bowtell DD: Inhibition of Siah ubiquitin ligase function. Oncogene. 28:289–296. 2009. View Article : Google Scholar : PubMed/NCBI
21	Lazar E, Benedek T, Korodi S, Rat N, Lo J and Benedek I: Stem cell-derived exosomes-an emerging tool for myocardial regeneration. World J Stem Cells. 10:106–115. 2018. View Article : Google Scholar : PubMed/NCBI
22	Messenger SW, Woo SS, Sun Z and Martin TFJ: A Ca²⁺ -stimulated exosome release pathway in cancer cells is regulated by Munc13-4. J Cell Biol. 217:2877–2890. 2018. View Article : Google Scholar : PubMed/NCBI
23	Ge R, Tan E, Sharghi-Namini S and Asada HH: Exosomes in cancer microenvironment and beyond: Have we overlooked these extracellular messengers? Cancer Microenviron. 5:323–332. 2012. View Article : Google Scholar : PubMed/NCBI
24	Yu S, Cao H, Shen B and Feng J: Tumor-derived exosomes in cancer progression and treatment failure. Oncotarget. 6:37151–37168. 2015. View Article : Google Scholar : PubMed/NCBI
25	Mathivanan S, Fahner CJ, Reid GE and Simpson RJ: ExoCarta 2012: Database of exosomal proteins, RNA and lipids. Nucleic Acids Res. 40:(Database Issue). D1241–D1244. 2012. View Article : Google Scholar : PubMed/NCBI
26	Théry C, Ostrowski M and Segura E: Membrane vesicles as conveyors of immune responses. Nat Rev Immunol. 9:581–593. 2009. View Article : Google Scholar : PubMed/NCBI
27	Bertoli G, Cava C and Castiglioni I: MicroRNAs as biomarkers for diagnosis, prognosis and theranostics in prostate cancer. Int J Mol Sci. 17:4212016. View Article : Google Scholar : PubMed/NCBI
28	Filella X and Foj L: miRNAs as novel biomarkers in the management of prostate cancer. Clin Chem Lab Med. 55:715–736. 2017. View Article : Google Scholar : PubMed/NCBI
29	Huang X, Yuan T, Liang M, Du M, Xia S, Dittmar R, Wang D, See W, Costello BA, Quevedo F, et al: Exosomal miR-1290 and miR-375 as prognostic markers in castration-resistant prostate cancer. Eur Urol. 67:33–41. 2015. View Article : Google Scholar : PubMed/NCBI
30	Foj L, Ferrer F, Serra M, Arévalo A, Gavagnach M, Giménez N and Filella X: Exosomal and non-exosomal urinary miRNAs in prostate cancer detection and prognosis. Prostate. 77:573–583. 2017. View Article : Google Scholar : PubMed/NCBI
31	Rucci N and Angelucci A: Prostate cancer and bone: The elective affinities. Biomed Res Int. 2014:1670352014. View Article : Google Scholar : PubMed/NCBI
32	Keller ET, Zhang J, Cooper CR, Smith PC, McCauley LK, Pienta KJ and Taichman RS: Prostate carcinoma skeletal metastases: Cross-talk between tumor and bone. Cancer Metastasis Rev. 20:333–349. 2001. View Article : Google Scholar : PubMed/NCBI
33	Mohla S: Under-investigated area in prostate cancer: Cross talk between the bone microenvironment and prostate cancer bone metastasis. J Cell Biochem. 91:684–685. 2004. View Article : Google Scholar : PubMed/NCBI
34	Paget S: The distribution of secondary growths in cancer of the breast. 1889. Cancer Metastasis Rev. 8:98–101. 1989.PubMed/NCBI
35	Engblom C, Pfirschke C, Zilionis R, Da Silva Martins J, Bos SA, Courties G, Rickelt S, Severe N, Baryawno N, Faget J, et al: Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neutrophils. Science. 358(pii): eaal50812017. View Article : Google Scholar : PubMed/NCBI
36	San Martin R, Pathak R, Jain A, Jung SY, Hilsenbeck SG, Piña-Barba MC, Sikora AG, Pienta KJ and Rowley DR: Tenascin-C and Integrin α9 mediate interactions of prostate cancer with the bone microenvironment. Cancer Res. 77:5977–5988. 2017. View Article : Google Scholar : PubMed/NCBI
37	Papaioannou G, Mirzamohammadi F and Kobayashi T: MicroRNAs involved in bone formation. Cell Mol Life Sci. 71:4747–4761. 2014. View Article : Google Scholar : PubMed/NCBI
38	Yun HM, Park KR, Quang TH, Oh H, Hong JT, Kim YC and Kim EC: 2,4,5-Trimethoxyldalbergiquinol promotes osteoblastic differentiation and mineralisation via the BMP and Wnt/β-catenin pathway. Cell Death Dis. 6:e18192015. View Article : Google Scholar : PubMed/NCBI
39	Lee HS, Jung EY, Bae SH, Kwon KH, Kim JM and Suh HJ: Stimulation of osteoblastic differentiation and mineralisation in MC3T3-E1 cells by yeast hydrolysate. Phytother Res. 25:716–723. 2011. View Article : Google Scholar : PubMed/NCBI