MicroRNA-129-5p suppresses cell proliferation, migration and invasion via targeting ROCK1 in osteosarcoma

Han,Chaofan; Wang,Wenbo

doi:10.3892/mmr.2018.8374

MicroRNA-129-5p suppresses cell proliferation, migration and invasion via targeting ROCK1 in osteosarcoma

Authors:
- Chaofan Han
- Wenbo Wang
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Affiliations: Department of Orthopedic Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: January 4, 2018 https://doi.org/10.3892/mmr.2018.8374
Pages: 4777-4784

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Abstract

Osteosarcoma (OS) is the most common primary malignancy of the bone in teenagers and accounts for 20‑35% of all malignant primary bone tumors. Increasing evidence shows that microRNAs (miRNAs) are abnormally expressed in several types of human cancer. miRNAs are necessary to maintain the malignant phenotype of cancer cells and can function as either tumor suppressors or oncogenes. The present study aimed to measure the expression of miRNA‑129‑5p (miR‑129‑5p) in OS, determine the effects of miR‑129‑5p on the malignant behaviors of OS cells, and elucidate the molecular mechanism underlying the oncogenesis and progression of OS. The expression levels of miR‑129‑5p in OS tissues and cell lines were measured using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. SAOS‑2 and U2OS cells were then transfected with miR‑129‑5p mimics or miR‑negative control. The effects of miR‑129‑5p on the proliferation, migration and invasion of SAOS‑2 and U2OS cells in vitro were then evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Transwell migration assay and invasion assays, respectively. In addition, bioinformatics analysis, a luciferase reporter assay, and RT‑qPCR and western blot analyses were used to examine whether Rho‑associated protein kinase 1 (ROCK1) was a direct target of miR‑129‑5p. The mRNA expression of ROCK1 in OS tissues was detected using RT‑qPCR analysis, and the biological roles of ROCK1 in OS cells were also evaluated. The results showed that miR‑129‑5p was significantly downregulated in the OS tissues and cell lines. The re‑expression of miR‑129‑5p suppressed the cell proliferation, migration and invasion of OS cells. In addition, ROCK1 was confirmed as a direct target of miR‑129‑5p. The mRNA expression of ROCK1 was high in OS tissues and inversely correlated with the expression of miR‑129‑5p. The downregulation of ROCK1 inhibited the proliferation, migration and invasion of OS cells. These findings suggested that miR‑129‑5p inhibited cell proliferation, migration and invasion in the development of OS via the negative regulation of ROCK1. The miR‑129‑5p/ROCK1 axis may serve as an efficient target in cancer therapy.

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1	Ottaviani G and Jaffe N: The epidemiology of osteosarcoma. Cancer Treat Res. 152:3–13. 2009. View Article : Google Scholar : PubMed/NCBI
2	Longhi A, Errani C, De Paolis M, Mercuri M and Bacci G: Primary bone osteosarcoma in the pediatric age: State of the art. Cancer Treat Rev. 32:423–436. 2006. View Article : Google Scholar : PubMed/NCBI
3	Jin J, Cai L, Liu ZM and Zhou XS: miRNA-218 inhibits osteosarcoma cell migration and invasion by down-regulating of TIAM1, MMP2 and MMP9. Asian Pac J Cancer Prev. 14:3681–3684. 2013. View Article : Google Scholar : PubMed/NCBI
4	Cho Y, Jung GH, Chung SH, Kim JY, Choi Y and Kim JD: Long-term survivals of stage IIb osteosarcoma: A 20-year experience in a single institution. Clin Orthop Surg. 3:48–54. 2011. View Article : Google Scholar : PubMed/NCBI
5	Bramer JA, van Linge JH, Grimer RJ and Scholten RJ: Prognostic factors in localized extremity osteosarcoma: A systematic review. Eur J Surg Oncol. 35:1030–1036. 2009. View Article : Google Scholar : PubMed/NCBI
6	Pierz KA, Womer RB and Dormans JP: Pediatric bone tumors: Osteosarcoma ewing's sarcoma and chondrosarcoma associated with multiple hereditary osteochondromatosis. J Pediatr Orthop. 21:412–418. 2001. View Article : Google Scholar : PubMed/NCBI
7	Poletajew S, Fus L and Wasiutynski A: Current concepts on pathogenesis and biology of metastatic osteosarcoma tumors. Ortop Traumatol Rehabil. 13:537–545. 2011.(In English, Polish). View Article : Google Scholar : PubMed/NCBI
8	Kong YW, Ferland-McCollough D, Jackson TJ and Bushell M: microRNAs in cancer management. Lancet Oncol. 13:e249–e258. 2012. View Article : Google Scholar : PubMed/NCBI
9	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
10	Kim VN: MicroRNA biogenesis: Coordinated cropping and dicing. Nat Rev Mol Cell Biol. 6:376–385. 2005. View Article : Google Scholar : PubMed/NCBI
11	Garzon R, Calin GA and Croce CM: MicroRNAs in cancer. Annu Rev Med. 60:167–179. 2009. View Article : Google Scholar : PubMed/NCBI
12	Liu J: Control of protein synthesis and mRNA degradation by microRNAs. Curr Opin Cell Biol. 20:214–221. 2008. View Article : Google Scholar : PubMed/NCBI
13	Carrington JC and Ambros V: Role of microRNAs in plant and animal development. Science. 301:336–338. 2003. View Article : Google Scholar : PubMed/NCBI
14	Tan X, Fan S, Wu W and Zhang Y: MicroRNA-26a inhibits osteosarcoma cell proliferation by targeting IGF-1. Bone Res. 3:150332015. View Article : Google Scholar : PubMed/NCBI
15	Zhao G, Dong L, Shi H, Li H, Lu X, Guo X and Wang J: MicroRNA-1207-5p inhibits hepatocellular carcinoma cell growth and invasion through the fatty acid synthase-mediated Akt/mTOR signalling pathway. Oncol Rep. 36:1709–1716. 2016. View Article : Google Scholar : PubMed/NCBI
16	Wu D, Niu X, Pan H, Zhou Y, Qu P and Zhou J: MicroRNA-335 is downregulated in bladder cancer and inhibits cell growth, migration and invasion via targeting ROCK1. Mol Med Rep. 13:4379–4385. 2016. View Article : Google Scholar : PubMed/NCBI
17	Croce CM: Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet. 10:704–714. 2009. View Article : Google Scholar : PubMed/NCBI
18	Yu Y, Zhao Y, Sun XH, Ge J, Zhang B, Wang X and Cao XC: Down-regulation of miR-129-5p via the Twist1-Snail feedback loop stimulates the epithelial-mesenchymal transition and is associated with poor prognosis in breast cancer. Oncotarget. 6:34423–34436. 2015. View Article : Google Scholar : PubMed/NCBI
19	Jiang Z, Wang H, Li Y, Hou Z, Ma N, Chen W, Zong Z and Chen S: miR-129-5p is down-regulated and involved in migration and invasion of gastric cancer cells by targeting interleukin-8. Neoplasma. 63:673–680. 2016. View Article : Google Scholar : PubMed/NCBI
20	Li J, Wang H, Ke H and Ni S: miR-129 regulates MMP9 to control metastasis of non-small cell lung cancer. Tumour Biol. 36:5785–5790. 2015. View Article : Google Scholar : PubMed/NCBI
21	Duan L, Hao X, Liu Z, Zhang Y and Zhang G: miR-129-5p is down-regulated and involved in the growth, apoptosis and migration of medullary thyroid carcinoma cells through targeting RET. FEBS Lett. 588:1644–1651. 2014. View Article : Google Scholar : PubMed/NCBI
22	Tan G, Cao X, Dai Q, Zhang B, Huang J, Xiong S, Zhang Yy, Chen W, Yang J and Li H: A novel role for microRNA-129-5p in inhibiting ovarian cancer cell proliferation and survival via direct suppression of transcriptional co-activators YAP and TAZ. Oncotarget. 6:8676–8686. 2015. View Article : Google Scholar : PubMed/NCBI
23	Karaayvaz M, Zhai H and Ju J: miR-129 promotes apoptosis and enhances chemosensitivity to 5-fluorouracil in colorectal cancer. Cell Death Dis. 4:e6592013. View Article : Google Scholar : PubMed/NCBI
24	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
25	Liu X, Choy E, Hornicek FJ, Yang S, Yang C, Harmon D, Mankin H and Duan Z: ROCK1 as a potential therapeutic target in osteosarcoma. J Orthop Res. 29:1259–1266. 2011. View Article : Google Scholar : PubMed/NCBI
26	Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W and Tuschl T: Identification of tissue-specific microRNAs from mouse. Curr Biol. 12:735–739. 2002. View Article : Google Scholar : PubMed/NCBI
27	Ma N, Chen F, Shen SL, Chen W, Chen LZ, Su Q, Zhang LJ, Bi J, Zeng WT, Li W, et al: MicroRNA-129-5p inhibits hepatocellular carcinoma cell metastasis and invasion via targeting ETS1. Biochem Biophys Res Commun. 461:618–623. 2015. View Article : Google Scholar : PubMed/NCBI
28	Zhai J, Qu S, Li X, Zhong J, Chen X, Qu Z and Wu D: miR-129 suppresses tumor cell growth and invasion by targeting PAK5 in hepatocellular carcinoma. Biochem Biophys Res Commun. 464:161–167. 2015. View Article : Google Scholar : PubMed/NCBI
29	Wang QY, Tang J, Zhou CX and Zhao Q: The down-regulation of miR-129 in breast cancer and its effect on breast cancer migration and motility. Sheng Li Xue Bao. 64:403–411. 2012.(In Chinese). PubMed/NCBI
30	Luo J, Chen J and He L: mir-129-5p Attenuates Irradiation-Induced Autophagy and Decreases Radioresistance of Breast Cancer Cells by Targeting HMGB1. Med Sci Monit. 21:4122–4129. 2015. View Article : Google Scholar : PubMed/NCBI
31	Zhang C, Zhang S, Zhang Z, He J, Xu Y and Liu S: ROCK has a crucial role in regulating prostate tumor growth through interaction with c-Myc. Oncogene. 33:5582–5591. 2014. View Article : Google Scholar : PubMed/NCBI
32	Rossman KL, Der CJ and Sondek J: GEF means go: Turning on RHO GTPases with guanine nucleotide-exchange factors. Nat Rev Mol Cell Biol. 6:167–180. 2005. View Article : Google Scholar : PubMed/NCBI
33	Patel RA, Forinash KD, Pireddu R, Sun Y, Sun N, Martin MP, Schönbrunn E, Lawrence NJ and Sebti SM: RKI-1447 is a potent inhibitor of the Rho-associated ROCK kinases with anti-invasive and antitumor activities in breast cancer. Cancer Res. 72:5025–5034. 2012. View Article : Google Scholar : PubMed/NCBI
34	Wang Y, Zhao W and Fu Q: miR-335 suppresses migration and invasion by targeting ROCK1 in osteosarcoma cells. Mol Cell Biochem. 384:105–111. 2013. View Article : Google Scholar : PubMed/NCBI