Abstract
The objective of this study was to explore the biological roles of microRNA-140 (miR-140) in tumor growth, migration, and metastasis of osteosarcoma (OS) in vivo and in vitro. Between 2007 and 2014, 47 cases of OS samples and normal bone tissue samples adjacent to OS were selected from our hospital. Tissue biopsies from OS patients were used to measure miR-140 levels to obtain a correlation between clinicopathological features and miR-140 expression. In vitro, MG63 human osteosarcoma cells were divided into four groups: blank group, miR-140 mimic group, miR-140 inhibitor group, and negative control (NC; empty plasmid) group. qRT-PCR was used to detect miR-140 expression, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell proliferation, flow cytometry was used to detect cell cycle distribution, and scratch migration assay was used to detect cell migration. In vivo, the relative expression of miR-140 level in OS tissue was lower than that in the adjacent normal bone tissue. miR-140 expression is inversely correlated with tumor size, Enneking stage, and tumor metastasis. In vitro, compared with blank group and NC group, relative miR-140 expression was increased, cell proliferation was inhibited, cell population in G0/G1 phase was increased, cell population in G2/M phase and S phases and proliferation index (PI), and cell migration distance were decreased in the miR-140 mimic group, but the relative expression and all the cell indexes were found opposite trend in the miR-140 inhibitor group. In conclusion, in vivo and vitro findings provided evidence that miR-140 could inhibit the growth, migration, and metastasis of OS cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Gorlick S, Barkauskas DA, Krailo M, Piperdi S, Sowers R, Gill J, et al. Her-2 expression is not prognostic in osteosarcoma; a children's oncology group prospective biology study. Pediatr Blood Cancer. 2014;61(9):1558–64.
Ritter J, Bielack SS. Osteosarcoma. Ann Oncol. 2010;21 Suppl 7:vii320–5.
Martin JW, Chilton-MacNeill S, Koti M, van Wijnen AJ, Squire JA, Zielenska M. Digital expression profiling identifies RUNX2, CDC5L, MDM2, RECQL4, and CDK4 as potential predictive biomarkers for neo-adjuvant chemotherapy response in paediatric osteosarcoma. PLoS One. 2014;9(5):e95843.
Lo WW, Wunder JS, Dickson BC, Campbell V, McGovern K, Alman BA, et al. Involvement and targeted intervention of dysregulated hedgehog signaling in osteosarcoma. Cancer. 2014;120(4):537–47.
Wang X, Du J, Gu P, Jin R, Lin X. Polymeric immunoglobulin receptor expression is correlated with poor prognosis in patients with osteosarcoma. Mol Med Rep. 2014;9(6):2105–10.
Lin F, Shen Z, Tang LN, Zheng SE, Sun YJ, Min DL, et al. KLF8 knockdown suppresses proliferation and invasion in human osteosarcoma cells. Mol Med Rep. 2014;9(5):1613–7.
Ji F, Zhang H, Wang Y, Li M, Xu W, Kang Y, et al. MicroRNA-133a, downregulated in osteosarcoma, suppresses proliferation and promotes apoptosis by targeting Bcl-xl and Mcl-1. Bone. 2013;56(1):220–6.
Cai H, Lin L, Cai H, Tang M, Wang Z. Combined microRNA-340 and ROCK1 mRNA profiling predicts tumor progression and prognosis in pediatric osteosarcoma. Int J Mol Sci. 2014;15(1):560–73.
Takata A, Otsuka M, Yoshikawa T, Kishikawa T, Hikiba Y, Obi S, et al. MicroRNA-140 acts as a liver tumor suppressor by controlling NF-kappaB activity by directly targeting DNA methyltransferase 1 (Dnmt1) expression. Hepatology. 2013;57(1):162–70.
Wolfson B, Eades G, Zhou Q. Roles of microRNA-140 in stem cell-associated early stage breast cancer. World J Stem Cells. 2014;6(5):591–7.
Kai Y, Peng W, Ling W, Jiebing H, Zhuan B. Reciprocal effects between microRNA-140-5p and ADAM10 suppress migration and invasion of human tongue cancer cells. Biochem Biophys Res Commun. 2014;448(3):308–14.
Song B, Wang Y, Xi Y, Kudo K, Bruheim S, Botchkina GI, et al. Mechanism of chemoresistance mediated by miR-140 in human osteosarcoma and colon cancer cells. Oncogene. 2009;28(46):4065–74.
Nakamura Y, Inloes JB, Katagiri T, Kobayashi T. Chondrocyte-specific microRNA-140 regulates endochondral bone development and targets Dnpep to modulate bone morphogenetic protein signaling. Mol Cell Biol. 2011;31(14):3019–28.
Yang J, Qin S, Yi C, Ma G, Zhu H, Zhou W, et al. MiR-140 is co-expressed with Wwp2-C transcript and activated by Sox9 to target Sp1 in maintaining the chondrocyte proliferation. FEBS Lett. 2011;585(19):2992–7.
Thampi S, Matthay KK, Boscardin WJ, Goldsby R, DuBois SG. Clinical features and outcomes differ between skeletal and extraskeletal osteosarcoma. Sarcoma. 2014;2014:902620.
Zhu J, Feng Y, Ke Z, Yang Z, Zhou J, Huang X, et al. Down-regulation of miR-183 promotes migration and invasion of osteosarcoma by targeting Ezrin. Am J Pathol. 2012;180(6):2440–51.
Hu H, Zhang Y, Cai XH, Huang JF, Cai L. Changes in microRNA expression in the MG-63 osteosarcoma cell line compared with osteoblasts. Oncol Lett. 2012;4(5):1037–42.
Kobayashi E, Hornicek FJ, Duan Z. MicroRNA involvement in osteosarcoma. Sarcoma. 2012;2012:359739.
Song B, Wang Y, Titmus MA, Botchkina G, Formentini A, Kornmann M, et al. Molecular mechanism of chemoresistance by miR-215 in osteosarcoma and colon cancer cells. Mol Cancer. 2010;9:96.
Ziyan W, Shuhua Y, Xiufang W, Xiaoyun L. MicroRNA-21 is involved in osteosarcoma cell invasion and migration. Med Oncol. 2011;28(4):1469–74.
Nie J, Liu L, Zheng W, Chen L, Wu X, Xu Y, et al. MicroRNA-365, down-regulated in colon cancer, inhibits cell cycle progression and promotes apoptosis of colon cancer cells by probably targeting Cyclin D1 and Bcl-2. Carcinogenesis. 2012;33(1):220–5.
Bian K, Fan J, Zhang X, Yang XW, Zhu HY, Wang L, et al. microRNA-203 leads to G1 phase cell cycle arrest in laryngeal carcinoma cells by directly targeting survivin. FEBS Lett. 2012;586(6):804–9.
Novello C, Pazzaglia L, Cingolani C, Conti A, Quattrini I, Manara MC, et al. miRNA expression profile in human osteosarcoma: role of miR-1 and miR-133b in proliferation and cell cycle control. Int J Oncol. 2013;42(2):667–75.
Mao JH, Zhou RP, Peng AF, Liu ZL, Huang SH, Long XH, et al. Microrna-195 suppresses osteosarcoma cell invasion and migration in vitro by targeting FASN. Oncol Lett. 2012;4(5):1125–9.
Jin J, Cai L, Liu ZM, Zhou XS. miRNA-218 inhibits osteosarcoma cell migration and invasion by down-regulating of TIAM1, MMP2 and MMP9. Asian Pac J Cancer Prev. 2013;14(6):3681–4.
Acknowledgments
We would like to thank our researchers for their hard work and reviewers for their valuable advice.
Conflicts of interest
None
Authors’ contributions
GR designed the study. SYF conceived and supervised the study. WMF performed the examination and the analysis. LJB performed the statistical analysis. JJL, WSH, WXL, and GQ interpreted the results. GR, JJL, and SYF drafted and revised the paper. All authors read and approved the final paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gu, R., Sun, YF., Wu, MF. et al. Biological roles of microRNA-140 in tumor growth, migration, and metastasis of osteosarcoma in vivo and in vitro. Tumor Biol. 37, 353–360 (2016). https://doi.org/10.1007/s13277-015-3801-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-015-3801-8