miRNA expression profile in human osteosarcoma: Role of miR-1 and miR-133b in proliferation and cell cycle control

Novello,Chiara; Pazzaglia,Laura; Cingolani,Chiara; Conti,Amalia; Quattrini,Irene; Manara,Maria  Cristina; Tognon,Mauro; Picci,Piero; Benassi,Maria  Serena

doi:10.3892/ijo.2012.1717

miRNA expression profile in human osteosarcoma: Role of miR-1 and miR-133b in proliferation and cell cycle control

Authors:
- Chiara Novello
- Laura Pazzaglia
- Chiara Cingolani
- Amalia Conti
- Irene Quattrini
- Maria Cristina Manara
- Mauro Tognon
- Piero Picci
- Maria Serena Benassi
View Affiliations

Affiliations: Laboratory of Experimental Oncology, Rizzoli Orthopaedic Institute, I-40136 Bologna, Italy, Department of Morphology and Embryology, Section of Cell Biology and Molecular Genetics, School of Medicine and Center of Biotechnology, University of Ferrara, I-44100 Ferrara, Italy
Published online on: November 28, 2012 https://doi.org/10.3892/ijo.2012.1717
Pages: 667-675

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

miRNA profile deregulation affecting downstream signaling pathways activates endpoints that represent potential biomarkers for prognosis and treatment of tumor patients. In the past 20 years conventional therapy for osteosarcoma (OS) reached a survival plateau, highlighting the need for new therapeutic approaches. In this study, microarray unsupervised and supervised analysis identified, respectively, 100 and 40 differentially expressed miRNAs in OS samples with different grades of malignancy compared to normal bone. When analyzing low-grade and high-grade OS by unsupervised analysis, 12 miRNAs were found to be differentially expressed. Real‑time PCR performed on a larger series of OS confirmed a significant lower expression of miR-1, miR‑133b and miR-378* in tumors with respect to control, also showing lower mRNA levels in 31 high-grade OS than in 25 low-grade and in metastatic versus non‑metastatic patients. We demonstrated that miR-1 and miR133b were downregulated in OS cell lines compared to normal osteoblasts. Secondly, by transfection with miRNA precursor molecules, we demonstrated that the ectopic expression of miR-1 and miR-133b in U2-OS cell lines significantly reduced cell proliferation and MET protein expression and negatively regulated cell invasiveness and motility in a short-term assay. Cell cycle distribution revealed block in G1 and delay of cell cycle progression associated with increased apoptosis in miR-1- and miR‑133b‑transfected cells, respectively. Our data assessed specific miRNA profiling deregulation in OS clinical samples and suggest that the expression of miR-1 and miR-133b may control cell proliferation and cell cycle through MET protein expression modulation.

View Figures

View References

1.	Gorlick R and Khanna C: Osteosarcoma. J Bone Miner Res. 25:683–691. 2010. View Article : Google Scholar
2.	Gougelet A, Pissaloux D, Besse A, et al: Micro-RNA profiles in osteosarcoma as predictive tool for ifosfamide response. Int J Cancer. 129:680–690. 2011. View Article : Google Scholar : PubMed/NCBI
3.	John B, Enright AJ, Aravin A, et al: Human MicroRNA targets. PloS Biol. 2:e3632004. View Article : Google Scholar
4.	Lagos-Quintana M, Rauhut R, Yalcin A, et al: Identification of tissue-specific microRNAs from mouse. Curr Biol. 12:735–39. 2002. View Article : Google Scholar : PubMed/NCBI
5.	Lee RC and Ambros V: An extensive class of small RNAs in Caenorhabditis elegans. Science. 294:862–64. 2001. View Article : Google Scholar : PubMed/NCBI
6.	Sempere LF, Freemantle S, Pitha-Rowe I, et al: Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation. Genome Biol. 5:R132004. View Article : Google Scholar
7.	Lim LP, Lau NC, Weinstein EG, et al: The microRNAs of Caenorhabditis elegans. Genes Dev. 17:991–1008. 2003.
8.	Esquela-Kerscher A and Slack FJ: Oncomirs - microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar
9.	Calin GA, Sevignani C, Dumitru CD, et al: Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA. 101:2999–3004. 2004. View Article : Google Scholar : PubMed/NCBI
10.	Wu W, Sun M, Zou GM and Chen J: MicroRNA and cancer: current status and prospective. Int J Cancer. 120:953–960. 2007. View Article : Google Scholar : PubMed/NCBI
11.	Montanini L, Lasagna L, Barili V, et al: MicroRNA cloning and sequencing in osteosarcoma cell lines: differential role of miR-93. Cell Oncol (Dordr). 35:29–41. 2012. View Article : Google Scholar : PubMed/NCBI
12.	He C, Xiong J, Hu X, et al: 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
13.	Kauppinen S, Vester B and Wengel J: Locked nucleic acid: high-affinity targeting of camplementary RNA for RNomics. Handb Exp Pharmacol. 173:405–422. 2006. View Article : Google Scholar : PubMed/NCBI
14.	Yang YH, Dudoit S, Luu P, et al: Normalization for cDNA microarray data :a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res. 30:e152002. View Article : Google Scholar : PubMed/NCBI
15.	Yan D, Dong Xda E, Chen X, et al: MicroRNA-1/206 targets c-Met and inhibits rhabdomyosarcoma development. J Biol Chem. 284:29596–29604. 2009. View Article : Google Scholar : PubMed/NCBI
16.	Hu G, Chen D, Li X, et al: miR-133b regulates the MET proto-oncogene and inhibits the growth of colorectal cancer cells in vitro and in vivo. Cancer Biol Ther. 10:190–197. 2010. View Article : Google Scholar : PubMed/NCBI
17.	Hattinger CM, Pasello M, Ferrari S, et al: Emergine drugs for high-grade osteosarcoma. Export Opin Emerg Drugs. 15:615–634. 2010. View Article : Google Scholar : PubMed/NCBI
18.	Zhang H, Cai X, Wang Y, et al: microRNA-143, down-regulated in osteosarcoma, promotes apoptosis and suppresses tumorigenicity by targeting Bcl-2. Oncol Rep. 24:1363–1369. 2010.PubMed/NCBI
19.	Ziyan W, Shuhua Y, Xiufang W and Xiaoyun L: MicroRNA-21 is involved in osteosarcoma cell invasion and migration. Med Oncol. 28:1469–1474. 2011. View Article : Google Scholar : PubMed/NCBI
20.	Song B, Wang Y, Titmus MA, et al: Molecular mechanism of chemoresistance by miR-215 in osteosarcoma and colon cancer cells. Mol Cancer. 9:962010. View Article : Google Scholar : PubMed/NCBI
21.	Bar N and Dikstein R: MiR-22 forms a regulatory loop in PTEN/AKT pathway and modulates signaling kinetics. PloS One. 27:e108592010. View Article : Google Scholar : PubMed/NCBI
22.	Huang Z, Huang S, Wang Q, et al: MicroRNA-95 promotes cell proliferation and targets sorting Nexin 1 in human colorectal carcinoma. Cancer Res. 71:2582–589. 2011. View Article : Google Scholar : PubMed/NCBI
23.	Kumarswamy R, Mudduluru G, Ceppi P, et al: MicroRNA-30a inhibits epithelial-to-mesenchymal transition by targeting Snail and is downregulated in non small cell lung cancer. Int J Cancer. 130:2044–2053. 2012. View Article : Google Scholar : PubMed/NCBI
24.	Lee DY, Deng Z, Wang CH and Yang BB: MicroRNA-378 promotes cell survival, tumor growth, and angiogenesis by targeting SuFu and Fus-1 expression. Proc Natl Acad Sci USA. 104:20350–20355. 2007. View Article : Google Scholar : PubMed/NCBI
25.	Chen JF, Mandel EM, Thomson JM, et al: The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nat Genet. 38:228–233. 2006. View Article : Google Scholar : PubMed/NCBI
26.	Carleton M, Cleary MA and Linsley PS: MicroRNAs and cell cycle regulation. Cell Cycle. 6:2127–2132. 2007. View Article : Google Scholar : PubMed/NCBI
27.	Crawford M, Batte K, Yu L, et al: MicroRNA 133B targets pro-survival molecules MCL-1 and BCL2L2 in lung cancer. Biochem Biophys Res Commun. 388:483–489. 2009. View Article : Google Scholar : PubMed/NCBI
28.	Taulli R, Bersani F, Foglizzo V, et al: The muscle-specific microRNA miR-206 blocks human rhabdomyosarcoma growth in xenotransplanted mice by promoting myogenic differentiation. J Clin Invest. 119:2366–2378. 2009.PubMed/NCBI