miR-221 increases osteosarcoma cell proliferation, invasion and migration partly through the downregulation of PTEN

Zhu,Jianwei; Liu,Fan; Wu,Quanming; Liu,Xiancheng

doi:10.3892/ijmm.2015.2352

miR-221 increases osteosarcoma cell proliferation, invasion and migration partly through the downregulation of PTEN

Authors:
- Jianwei Zhu
- Fan Liu
- Quanming Wu
- Xiancheng Liu
View Affiliations

Affiliations: Department of Orthopedics, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China, Department of Oncology, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
Published online on: September 22, 2015 https://doi.org/10.3892/ijmm.2015.2352
Pages: 1377-1383

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Increasing evidence has demonstrated that microRNAs (miRNAs or miRs) are involved in cancer initiation and progression. Previous studies have indicated that miR-221 is one of the most consistently overexpressed miRNAs in multiple types of cancer. However, the role of miR-221 in osteosarcoma carcinogenesis and progression is not yet fully understood. Thus, the aim of the present study was to examine the expression of miR-221 in osteosarcoma and to determine the effects of miR-221 on the biological behavior of osteosarcoma cells. RT-qPCR revealed that the expression of miR-221 was significantly upregulated in the osteosarcoma tissues and osteosarcoma cell lines (p<0.05). In order to explore the role of miR-221 in osteosarcoma, the expression of miR-221 in the human osteosarcoma cell line MG‑63 was upregulated or downregulated by transfection with miR-221 mimic or miR-221 inhibitor, respectively. The results from RT-qPCR revealed that we had successfully generated MG‑63 cells in which miR-221 was either overexpressed or depleted. To investigate the effects of miR-221 on osteosarcoma cell proliferation, invasion and migration, a tetrazolium-based colorimetric assay, propidium iodide (PI) staining, a transwell migration assay and a wound healing assay were used in the present study. The results revealed that the proliferation, invasion and migration ability of the MG‑63 cells in which miR-221 was overexpressed was enhanced, and the proliferation, invasion and migration ability of the MG‑63 cells in which miR-221 was depleted was suppressed. The correlation between miR-221 and phosphatase and tensin homolog (PTEN) expression was investigated by RT-qPCR and western blot analysis. The results revealed that the downregulation of miR-221 significantly increased the expression of PTEN, whereas the upregulation of miR-221 significantly reduced the expression of PTEN. Taken together, our results suggest that miR-221 enhances the proliferation, invasion and migration ability of osteosarcoma cells partly by suppressing PTEN.

View Figures

View References

1	Endo-Munoz L, Evdokiou A and Saunders NA: The role of osteoclasts and tumour-associated macrophages in osteosarcoma metastasis. Biochim Biophys Acta. 1826.434–442. 2012.
2	Lamora A, Talbot J, Bougras G, Amiaud J, Leduc M, Chesneau J, Taurelle J, Stresing V, Le Deley MC, Heymann MF, et al: Overexpression of smad7 blocks primary tumor growth and lung metastasis development in osteosarcoma. Clin Cancer Res. 20:5097–5112. 2014. View Article : Google Scholar : PubMed/NCBI
3	Harting MT and Blakely ML: Management of osteosarcoma pulmonary metastases. Semin Pediatr Surg. 15:25–29. 2006. View Article : Google Scholar : PubMed/NCBI
4	Jia S and Li B: Osteosarcoma of the jaws: case report on synchronous multicentric osteosarcomas. J Clin Diagn Res. 8:ZD01–ZD03. 2014.PubMed/NCBI
5	He JP, Hao Y, Wang XL, Yang XJ, Shao JF, Guo FJ and Feng JX: Review of the molecular pathogenesis of osteosarcoma. Asian Pac J Cancer Prev. 15:5967–5976. 2014. View Article : Google Scholar : PubMed/NCBI
6	Poletajew S, Fus L and Wasiutyński A: Current concepts on pathogenesis and biology of metastatic osteosarcoma tumors. Ortop Traumatol Rehabil. 13:537–545. 2011. View Article : Google Scholar
7	Egas-Bejar D, Anderson PM, Agarwal R, Corrales-Medina F, Devarajan E, Huh WW, Brown RE and Subbiah V: Theranostic profiling for actionable aberrations in advanced high risk osteosarcoma with aggressive biology breveals high molecular diversity: the human fingerprint hypothesis. Oncoscience. 1:167–179. 2014. View Article : Google Scholar : PubMed/NCBI
8	Wu CW, Cheng YW, Hsu NY, Yeh KT, Tsai YY, Chiang CC, Wang WR and Tung JN: MiRNA-221 negatively regulated downstream p27Kip1 gene expression involvement in pterygium pathogenesis. Mol Vis. 20:1048–1056. 2014.PubMed/NCBI
9	Wolter JM, Kotagama K, Pierre-Bez AC, Firago M and Mangone M: 3′LIFE: a functional assay to detect miRNA targets in high-throughput. Nucleic Acids Res. 42:e1322014. View Article : Google Scholar
10	Gaál Z and Oláh E: MicroRNA-s and their role in malignant hematologic diseases. Orv Hetil. 153:2051–2059. 2012.In Hungarian. View Article : Google Scholar
11	Lin S, Pan L, Guo S, Wu J, Jin L, Wang JC and Wang S: Prognostic role of microRNA-181a/b in hematological malignancies: a meta-analysis. PLoS One. 8:e595322013. View Article : Google Scholar : PubMed/NCBI
12	De Sarkar N, Roy R, Mitra JK, Ghose S, Chakraborty A, Paul RR, Mukhopadhyay I and Roy B: a quest for miRNA bio-marker: a track back approach from gingivo buccal cancer to two different types of precancers. PLoS One. 9:e1048392014. View Article : Google Scholar : PubMed/NCBI
13	Xu C, Ping Y and Li X, Zhao H, Wang L, Fan H, Xiao Y and Li X: Prioritizing candidate disease miRNAs by integrating phenotype associations of multiple diseases with matched miRNA and mRNA expression profiles. Mol Biosyst. 10:2800–2809. 2014. View Article : Google Scholar : PubMed/NCBI
14	Ye S, Yang L, Zhao X, Song W, Wang W and Zheng S: Bioinformatics method to predict two regulation mechanism: TF-miRNA-mRNA and lncRNA-miRNA-mRNA in pancreatic cancer. Cell Biochem Biophys. 70:1849–1858. 2014. View Article : Google Scholar : PubMed/NCBI
15	Kushwaha D, Ramakrishnan V, Ng K, Steed T, Nguyen T, Futalan D, Akers JC, Sarkaria J, Jiang T, Chowdhury D, et al: A genome-wide miRNA screen revealed miR-603 as a MGMT-regulating miRNA in glioblastomas. Oncotarget. 5:4026–4039. 2014. View Article : Google Scholar : PubMed/NCBI
16	Kalinowski FC, Brown RA, Ganda C, Giles KM, Epis MR, Horsham J and Leedman PJ: microRNA-7: a tumor suppressor miRNA with therapeutic potential. Int J Biochem Cell Biol. 54:312–317. 2014. View Article : Google Scholar : PubMed/NCBI
17	Garofalo M, Quintavalle C, Romano G, Croce CM and Condorelli G: miR221/222 in cancer: their role in tumor progression and response to therapy. Curr Mol Med. 12:27–33. 2012. View Article : Google Scholar
18	le Sage C, Nagel R, Egan DA, Schrier M, Mesman E, Mangiola A, Anile C, Maira G, Mercatelli N, Ciafrè SA, et al: Regulation of the p27^Kip1 tumor suppressor by miR-221 and miR-222 promotes cancer cell proliferation. EMBO J. 26:3699–3708. 2007. View Article : Google Scholar : PubMed/NCBI
19	Fornari F, Gramantieri L, Ferracin M, Veronese A, Sabbioni S, Calin GA, Grazi GL, Giovannini C, Croce CM, Bolondi L and Negrini M: MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma. Oncogene. 27:5651–5661. 2008. View Article : Google Scholar : PubMed/NCBI
20	Miller TE, Ghoshal K, Ramaswamy B, Roy S, Datta J, Shapiro CL, Jacob S and Majumder S: MicroRNA-221/222 confers tamoxifen resistance in breast cancer by targeting p27^Kip1. J Biol Chem. 283:29897–29903. 2008. View Article : Google Scholar : PubMed/NCBI
21	Garofalo M, Di Leva G, Romano G, Nuovo G, Suh SS, Ngankeu A, Taccioli C, Pichiorri F, Alder H, Secchiero P, et al: miR-221&222 regulate TRAIL resistance and enhance tumorigenicity through PTEN and TIMP3 downregulation. Cancer Cell. 16:498–509. 2009. View Article : Google Scholar : PubMed/NCBI
22	Howe EN, Cochrane DR and Richer JK: The miR-200 and miR-221/222 microRNA families: opposing effects on epithelial identity. J Mammary Gland Biol Neoplasia. 17:65–77. 2012. View Article : Google Scholar : PubMed/NCBI
23	Galardi S, Mercatelli N, Giorda E, Massalini S, Frajese GV, Ciafrè SA and Farace MG: miR-221 and miR-222 expression affects the proliferation potential of human prostate carcinoma cell lines by targeting p27Kip1. J Biol Chem. 282:23716–23724. 2007. View Article : Google Scholar : PubMed/NCBI
24	He XX, Guo AY, Xu CR, Chang Y, Xiang GY, Gong J, Dan ZL, Tian DA, Liao JZ and Lin JS: Bioinformatics analysis identifies miR-221 as a core regulator in hepatocellular carcinoma and its silencing suppresses tumor properties. Oncol Rep. 32:1200–1210. 2014.PubMed/NCBI
25	Liu S, Sun X, Wang M, Hou Y, Zhan Y, Jiang Y, Liu Z, Cao X, Chen P, Liu Z, et al: A microRNA miR-221- and miR-222-mediated feedback loop, via PDLIM2, maintains constitutive activation of nuclear factor kappaB and STAT3 in colorectal cancer cells. Gastroenterology. 147:847–859. 2014. View Article : Google Scholar
26	Gan R, Yang Y, Yang X, Zhao L, Lu J and Meng QH: Downregulation of miR-221/222 enhances sensitivity of breast cancer cells to tamoxifen through upregulation of TIMP3. Cancer Gene Ther. 21:290–296. 2014. View Article : Google Scholar : PubMed/NCBI
27	Felli N, Fontana L, Pelosi E, Botta R, Bonci D, Facchiano F, Liuzzi F, Lulli V, Morsilli O, Santoro S, et al: MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation. Proc Natl Acad Sci USA. 102:18081–18086. 2005. View Article : Google Scholar : PubMed/NCBI
28	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
29	Ye X, Bai W, Zhu H, Zhang X, Chen Y, Wang L, Yang A, Zhao J and Jia L: MiR-221 promotes trastuzumab-resistance and metastasis in HER2-positive breast cancers by targeting PTEN. BMB Rep. 47:268–273. 2014. View Article : Google Scholar :
30	De Tullio G, De Fazio V, Sgherza N, Minoia C, Serratì S, Merchionne F, Loseto G, Iacobazzi A, Rana A, Petrillo P, et al: Challenges and opportunities of microRNAs in lymphomas. Molecules. 19:14723–14781. 2014. View Article : Google Scholar : PubMed/NCBI
31	Xu Y, Huang Z and Liu Y: Reduced miR-125a-5p expression is associated with gastric carcinogenesis through the targeting of E2F3. Mol Med Rep. 10:2601–2608. 2014.PubMed/NCBI
32	Tao K, Yang J, Guo Z, Hu Y, Sheng H, Gao H and Yu H: Prognostic value of miR-221-3p, miR-342-3p and miR-491-5p expression in colon cancer. Am J Transl Res. 6:391–401. 2014.PubMed/NCBI
33	Wei Y, Lai X, Yu S, Chen S, Ma Y, Zhang Y, Li H, Zhu X, Yao L and Zhang J: Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells. Breast Cancer Res Treat. 147:423–431. 2014. View Article : Google Scholar : PubMed/NCBI
34	Yang X, Yang Y, Gan R, Zhao L, Li W, Zhou H, Wang X, Lu J and Meng QH: Down-regulation of mir-221 and mir-222 restrain prostate cancer cell proliferation and migration that is partly mediated by activation of SIRT1. PLoS One. 9:e988332014. View Article : Google Scholar : PubMed/NCBI
35	Falkenberg N, Anastasov N, Rappl K, Braselmann H, Auer G, Walch A, Huber M, Höfig I, Schmitt M, Höfler H, et al: MiR-221/-222 differentiate prognostic groups in advanced breast cancers and influence cell invasion. Br J Cancer. 109:2714–2723. 2013. View Article : Google Scholar : PubMed/NCBI
36	Nassirpour R, Mehta PP, Baxi SM and Yin MJ: miR-221 promotes tumorigenesis in human triple negative breast cancer cells. PLoS One. 8:e621702013. View Article : Google Scholar : PubMed/NCBI
37	Chen G, Dang YW and Luo DZ: Effect of miR-221 on the viability and apoptosis of hepatocellular carcinoma HepG2 cells. Zhonghua Gan Zang Bing Za Zhi. 19:582–587. 2011.In Chinese. PubMed/NCBI
38	Hsieh TH, Chien CL, Lee YH, Lin CI, Hsieh JY, Chao ME, Liu DJ, Chu SS, Chen W, Lin SC, et al: Downregulation of SUN2, a novel tumor suppressor, mediates miR-221/222-induced malignancy in central nervous system embryonal tumors. Carcinogenesis. 35:2164–2174. 2014. View Article : Google Scholar : PubMed/NCBI
39	Sarkar S, Dubaybo H, Ali S, Goncalves P, Kollepara SL, Sethi S, Philip PA and Li Y: Down-regulation of miR-221 inhibits proliferation of pancreatic cancer cells through up-regulation of PTEN, p27^kip1, p57^kip2, and PUMA. Am J Cancer Res. 3:465–477. 2013.
40	Hwang MS, Yu N, Stinson SY, Yue P, Newman RJ, Allan BB and Dornan D: miR-221/222 targets adiponectin receptor 1 to promote the epithelial-to-mesenchymal transition in breast cancer. PLoS One. 8:e665022013. View Article : Google Scholar : PubMed/NCBI
41	Sun T, Wang X, He HH, Sweeney CJ, Liu SX, Brown M, Balk S, Lee GS and Kantoff PW: MiR-221 promotes the development of androgen independence in prostate cancer cells via downregulation of HECTD2 and RAB1A. Oncogene. 33:2790–2800. 2014. View Article : Google Scholar :
42	Jikuzono T, Kawamoto M, Yoshitake H, Kikuchi K, Akasu H, Ishikawa H, Hirokawa M, Miyauchi A, Tsuchiya S, Shimizu K and Takizawa T: The miR-221/222 cluster, miR-10b and miR-92a are highly upregulated in metastatic minimally invasive follicular thyroid carcinoma. Int J Oncol. 42:1858–1868. 2013.PubMed/NCBI
43	Acunzo M, Visone R, Romano G, Veronese A, Lovat F, Palmieri D, Bottoni A, Garofalo M, Gasparini P, Condorelli G, et al: miR-130a targets MET and induces TRAIL-sensitivity in NSCLC by downregulating miR-221 and 222. Oncogene. 31:634–642. 2012.
44	Wang H, Xu C, Kong X, Li X, Kong X, Wang Y, Ding X and Yang Q: Trail resistance induces epithelial-mesenchymal transition and enhances invasiveness by suppressing PTEN via miR-221 in breast cancer. PLoS One. 9:e990672014. View Article : Google Scholar : PubMed/NCBI