miR-34a inhibits the metastasis of osteosarcoma cells by repressing the expression of CD44

Zhao,Haien; Ma,Baoan; Wang,Yucai; Han,Tao; Zheng,Lianhe; Sun,Cong; Liu,Tao; Zhang,Yinglong; Qiu,Xiuchun; Fan,Qingyu

doi:10.3892/or.2013.2234

miR-34a inhibits the metastasis of osteosarcoma cells by repressing the expression of CD44

Authors:
- Haien Zhao
- Baoan Ma
- Yucai Wang
- Tao Han
- Lianhe Zheng
- Cong Sun
- Tao Liu
- Yinglong Zhang
- Xiuchun Qiu
- Qingyu Fan
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Affiliations: Department of Orthopaedic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi 710038, P.R. China
Published online on: January 11, 2013 https://doi.org/10.3892/or.2013.2234
Pages: 1027-1036

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Abstract

Osteosarcoma is the most common type of malignant bone tumor in children and adolescents and approximately 30% of patients develop lung metastasis, which is the leading cause of mortality. In this study, we investigated the role of miR-34a in the invasion and metastasis of osteosarcoma cells by examining its expression level and functional pattern in these cells. miR-34a mimics were transfected into the highly metastatic subline, F5M2, and into the F4 subline with low metastatic potential of the paired human osteosarcoma cell line, SOSP‑9607. Cell viability patterns, cell migration and alterations in gene expression levels were assessed by real-time PCR, and changes in protein levels were assessed by immunocytochemistry and western blot analysis. The ectopic overexpression of miR-34a significantly inhibited the migration and invasive ability of osteosarcoma cells by repressing the expression of CD44. These data suggest that miR-34a plays a tumor suppressor role in the metastasis of osteosarcoma cells by repressing the expression of CD44. Of note, studies have also suggested that the CD44 protein correlates with the metastatic potential of several malignant tumors. Therefore, it can be concluded that through the inhibition of CD44 expression levels, miR-34a plays a significant role in the migration and invasion of osteosarcoma cells.

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1	Bentzen SM, Poulsen HS, Kaae S, Jensen OM, Johansen H, Mouridsen HT, Daugaard S and Arnoldi C: Prognostic factors in osteosarcomas, a regression analysis. Cancer. 62:194–202. 1988. View Article : Google Scholar : PubMed/NCBI
2	Davis AM, Bell RS and Goodwin PJ: Prognostic factors in osteosarcoma: a critical review. J Clin Oncol. 12:423–431. 1994.PubMed/NCBI
3	Mankin HJ, Hornicek FJ, Rosenberg AE, Harmon DC and Gebhardt MC: Survival data for 648 patients with osteosarcoma treated at one institution. Clin Orthop Relat Res. 429:286–291. 2004. View Article : Google Scholar : PubMed/NCBI
4	Filipowicz W, Bhattacharyya SN and Sonenberg N: Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet. 9:102–114. 2008. View Article : Google Scholar : PubMed/NCBI
5	Medina PP and Slack FJ: microRNAs and cancer: an overview. Cell Cycle. 7:2485–2492. 2008. View Article : Google Scholar : PubMed/NCBI
6	Pillai RS, Bhattacharyya SN and Filipowiczm W: Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol. 17:118–126. 2007. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
8	Zhang B, Pan X, Cobb GP and Anderson TA: microRNAs as oncogenes and tumor suppressors. Dev Biol. 302:1–12. 2007. View Article : Google Scholar : PubMed/NCBI
9	Bartel DP: MicroRNAs: target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
10	Caldas C and Brenton JD: Sizing up miRNAs as cancer genes. Nat Med. 11:712–714. 2005. View Article : Google Scholar : PubMed/NCBI
11	He L, He X, Lim LP, de Stanchina E, Xuan Z, et al: A microRNA component of the p53 tumour suppressor network. Nature. 447:1130–1134. 2007. View Article : Google Scholar : PubMed/NCBI
12	Raver-Shapira N, Marciano E, Meiri E, Spector Y, Rosenfeld N, et al: Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. Mol Cell. 26:731–743. 2007. View Article : Google Scholar : PubMed/NCBI
13	Welch C, Chen Y and Stallings RL: MicroRNA-34a functions as a potential tumor suppressor by inducing apoptosis in neuroblastoma cells. Oncogene. 26:5017–5022. 2007. View Article : Google Scholar : PubMed/NCBI
14	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
15	Sun F, Fu H, Liu Q, Tie Y, Zhu J, et al: Downregulation of CCND1 and CDK6 by miR-34a induces cell cycle arrest. FEBS Lett. 582:1564–1568. 2005. View Article : Google Scholar : PubMed/NCBI
16	Wei JS, Song YK, Durinck S, Chen QR, Cheuk AT, et al: The MYCN oncogene is a direct target of miR-34a. Oncogene. 27:5204–5213. 2008. View Article : Google Scholar : PubMed/NCBI
17	Yamakuchi M, Ferlito M and Lowenstein CJ: miR-34a repression of SIRT1 regulates apoptosis. Proc Natl Acad Sci USA. 105:13421–13426. 2008. View Article : Google Scholar : PubMed/NCBI
18	Chen X, Yang TT, Wang W, Sun HH, Ma BA, Li CX, Ma Q, Yu Z and Fan QY: Establishment and characterization of human osteosarcoma cell lines with different pulmonary metastatic potentials. Cytotechnology. 61:37–44. 2009. View Article : Google Scholar : PubMed/NCBI
19	He C, Xiong J, Xu X, Lu W, Liu L, Xiao D and Wang D: Functional elucidation of MiR-34 in osteosarcoma cells and primary tumor samples. Biochem Biophys Res Commun. 388:35–40. 2009. View Article : Google Scholar : PubMed/NCBI
20	Guo Y, Li S, Qu J, Wang S, Dang Y, et al: MiR-34a inhibits lymphatic metastasis potential of mouse hepatoma cells. Mol Cell Biochem. 354:275–282. 2011. View Article : Google Scholar : PubMed/NCBI
21	Liu C, Kelnar K, Liu B, Chen X, Calhoun-Davis T, et al: The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nat Med. 17:211–215. 2011. View Article : Google Scholar : PubMed/NCBI
22	Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP and Burge CB: Prediction of mammalian microRNA targets. Cell. 115:787–798. 2003. View Article : Google Scholar : PubMed/NCBI
23	Krek A, Grün D, Poy MN, Wolf R, Rosenberg L, et al: Combinatorial microRNA target predictions. Nat Genet. 37:495–500. 2005. View Article : Google Scholar
24	John B, Enright AJ, Aravin A, Tuschl T, Sander C and Marks DS: Human MicroRNA targets. PLoS Biol. 2:e2642004. View Article : Google Scholar
25	Coltella N, Manara MC, Cerisano V, Trusolino L, et al: Role of the MET/HGF receptor in proliferation and invasive behavior of osteosarcoma. FASEB J. 17:1162–1164. 2003.PubMed/NCBI
26	Tavazoie SF, Alarcón C, Oskarsson T, Padua D, Wang Q, et al: Endogenous human microRNAs that suppress breast cancer metastasis. Nature. 451:147–152. 2008. View Article : Google Scholar : PubMed/NCBI
27	Leite KR, Sousa-Canavez JM, Reis ST, Tomiyama AH, Camara-Lopes LH, Sañudo A, Antunes AA and Srougi M: Change in expression of miR-let7c, miR-100, and miR-218 from high grade localized prostate cancer to metastasis. Urol Oncol. 29:265–269. 2009. View Article : Google Scholar : PubMed/NCBI
28	Mazar J, DeBlasio D, Govindarajan SS, Zhang S and Perera RJ: Epigenetic regulation of microRNA-375 and its role in melanoma development in humans. FEBS Lett. 585:2467–2476. 2011. View Article : Google Scholar : PubMed/NCBI
29	Kumar MS, Lu J, Mercer KL, Golub TR and Jacks T: Impaired microRNA processing enhances cellular transformation and tumorigenesis. Nat Genet. 39:673–677. 2007. View Article : Google Scholar : PubMed/NCBI
30	Tan S, Li R, Ding K, Lobie PE and Zhu T: miR-198 inhibits migration and invasion of hepatocellular carcinoma cells by targeting the HGF/c-MET pathway. FEBS Lett. 585:2229–2234. 2011. View Article : Google Scholar : PubMed/NCBI
31	Wu L, Cai C, Wang X, Liu M, Li X and Tang H: MicroRNA-142-3p, a new regulator of RAC1, suppresses the migration and invasion of hepatocellular carcinoma cells. FEBS Lett. 585:1322–1330. 2011. View Article : Google Scholar : PubMed/NCBI
32	Cozzolino AM, Pedace L, Castori M, De Simone P, Preziosi N, et al: Analysis of the miR-34a locus in 62 patients with familial cutaneous melanoma negative for CDKN2A/CDK4 screening. Fam Cancer. 11:201–208. 2012. View Article : Google Scholar : PubMed/NCBI
33	Yan D, Zhou X, Chen X, Hu D, Dong X, et al: MicroRNA-34a Inhibits uveal melanoma cell proliferation and migration through downregulation of c-Met. Invest Ophthalmol Vis Sci. 50:1559–1565. 2009. View Article : Google Scholar : PubMed/NCBI
34	Zhao T, Li J and Chen AF: MicroRNA-34a induces endothelial progenitor cell senescence and impedes its angiogenesis via suppressing silent information regulator 1. Am J Physiol Endocrinol Metab. 299:E110–E116. 2010. View Article : Google Scholar : PubMed/NCBI
35	Pang RT, Leung CO, Ye TM, Liu W, Chiu PC, et al: MicroRNA-34a suppresses invasion through downregulation of Notch1 and Jagged1 in cervical carcinoma and choriocarcinoma cells. Carcinogenesis. 31:1037–1044. 2010. View Article : Google Scholar : PubMed/NCBI
36	Doench JG and Sharp PA: Specificity of microRNA target selection in translational repression. Genes Dev. 18:504–511. 2004. View Article : Google Scholar : PubMed/NCBI
37	Aruffo A, Stamenkovic I, Melnick M, Underhill CB and Seed B: CD44 is the principal cell surface receptor for hyaluronate. Cell. 61:1303–1313. 1990. View Article : Google Scholar : PubMed/NCBI
38	Ponta H, Sherman L and Herrlich PA: CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol. 4:33–45. 2003. View Article : Google Scholar : PubMed/NCBI
39	Marhaba R and Zöller M: CD44 in cancer progression: adhesion, migration and growth regulation. J Mol Histol. 35:211–231. 2004. View Article : Google Scholar : PubMed/NCBI
40	Gotte M and Yip GW: Heparanase, hyaluronan, and CD44 in cancers: a breast carcinoma perspective. Cancer Res. 66:10233–10237. 2006. View Article : Google Scholar : PubMed/NCBI
41	Olsson E, Honeth G, Bendahl PO, Saal LH, Gruvberger-Saal S, Ringnér M, et al: CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers. BMC Cancer. 11:4182011. View Article : Google Scholar : PubMed/NCBI
42	Heyse TJ, Malcherczyk D, Moll R, Timmesfeld N, Wapelhorst J, et al: CD44: survival and metastasis in chondrosarcoma. Osteoarthritis Cartilage. 18:849–856. 2010. View Article : Google Scholar : PubMed/NCBI
43	Chen KL, Pan F, Jiang H, Chen JF, Pei L, Xie FW and Liang HJ: Highly enriched CD133⁺CD44⁺ stem-like cells with CD133⁺CD44^high metastatic subset in HCT116 colon cancer cells. Clin Exp Metastasis. 28:751–763. 2011.
44	Zhang C, Li C, He F, Cai Y and Yang H: Identification of CD44⁺CD24⁺ gastric cancer stem cells. J Cancer Res Clin Oncol. 137:1679–1686. 2011.PubMed/NCBI
45	Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, et al: Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells. 27:1006–1020. 2009. View Article : Google Scholar : PubMed/NCBI
46	Afify A, Purnell P and Nguyen L: Role of CD44s and CD44v6 on human breast cancer cell adhesion, migration, and invasion. Exp Mol Pathol. 86:95–100. 2009. View Article : Google Scholar : PubMed/NCBI
47	Kim HS, Park YB, Oh JH, Jeong J, Kim CJ and Lee SH: Expression of CD44 isoforms correlates with the metastatic potential of osteosarcoma. Clin Orthop Relat Res. 396:184–190. 2002. View Article : Google Scholar : PubMed/NCBI
48	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
49	Chen R, Nishimura MC, Bumbaca SM, et al: A hierarchy of self-renewing tumor-initiating cell types in glioblastoma. Cancer Cell. 17:362–375. 2010. View Article : Google Scholar : PubMed/NCBI
50	Jin L, Hope KJ, Zhai Q, Smadja-Joffe F and Dick JE: Targeting of CD44 eradicates human acute myeloid leukemic stem cells. Nat Med. 12:1167–1174. 2006. View Article : Google Scholar : PubMed/NCBI
51	Ji Q, Hao X, Zhang M, Tang W, Yang M, et al: MicroRNA miR-34 inhibits human pancreatic cancer tumor-initiating cells. PLoS One. 4:e68162009. View Article : Google Scholar : PubMed/NCBI
52	Wiggins JF, Ruffino L, Kelnar K, Omotola M, Patrawala L, et al: Development of a lung cancer therapeutic based on the tumor suppressor microRNA-34. Cancer Res. 70:5923–5930. 2010. View Article : Google Scholar : PubMed/NCBI
53	Huang N, Lin J, Ruan J, Su N, Qing R, Liu F, He B, Lv C, Zheng D and Luo R: MiR-219-5p inhibits hepatocellular carcinoma cell proliferation by targeting glypican-3. FEBS Lett. 586:884–891. 2012. View Article : Google Scholar : PubMed/NCBI
54	Kang HW, Wang F, Wei Q, Zhao YF, Liu M, Li X and Tang H: miR-20a promotes migration and invasion by regulating TNKS2 in human cervical cancer cells. FEBS Lett. 586:897–904. 2012. View Article : Google Scholar : PubMed/NCBI
55	Wu L, Li H, Jia CY, Cheng W, Yu M, Peng M, Zhu Y, Zhao Q, Dong YW, Shao K, Wu A and Wu XZ: MicroRNA-223 regulates FOXO1 expression and cell proliferation. FEBS Lett. 586:1038–1043. 2012. View Article : Google Scholar : PubMed/NCBI