CDA-2 induces cell differentiation through suppressing Twist/SLUG signaling via miR-124 in glioma
- 603 Downloads
Cell differentiation agent-2 (CDA-2) is an extraction from healthy human urine consisting of primary organic acids and peptides, and it has been demonstrated to inhibit growth and induce differentiation in glioma and other cell lines. But the mechanism of CDA-2 remains unclear. In this study, we demonstrated that CDA-2 inhibited cell growth and induced differentiation of glioma cells, accompanied with decreased expression of SLUG, Twist and Vimentin in both SWO-38 and U251 cell lines. Overexpression of SLUG or Twist greatly eliminated the efficiency of CDA-2 in inducing differentiation. Further study showed that CDA-2 treatment resulted in great changed microRNAs (miRNAs) detected by quantitative PCR, in which miR-124 was one of the most changed miRNAs and its level was increased by fourfold. The result of miRNA target prediction showed that miR-124 could regulate hundreds of genes which were relative to cell differentiation, such as SLUG, Vimentin, actin cytoskeleton, focal adhesion, tight junction. Inhibition of miR-124 up-regulated SLUG, Twist and Vimentin proteins, and partly eliminated the function of CDA-2 on these mesenchymal markers. Our findings demonstrated for the first time that CDA-2 induced cell differentiation through suppressing Twist and SLUG via miR-124 in glioma cells.
KeywordsCDA-2 miR-124 SLUG Twist Glioma
This research was supported by Foundation of Science and Technology Department of Guangdong Province, China (No. 2011B080701021).
- 1.Yao CJ, Lai GM, Chan CF, Yang YY, Liu FC, Chuang SE (2005) Differentiation of pheochromocytoma PC12 cells induced by human urine extract and the involvement of the extracellular signal-regulated kinase signaling pathway. J Altern Complement Med 11:903–908. doi: 10.1089/acm.2005.11.903 PubMedCrossRefGoogle Scholar
- 2.Xin Y, Xu KS (2001) HPLC determination of peptide’s molecular weight in Niaoduosuantai (CDA-II) injection. Chin J Pharm Anal 21:191–193Google Scholar
- 4.Wang HY, Zhong XY, Tu YS, Liu ZZ (2006) Effect of CDA-II on the differentiation and inhibition of human glioma cell lines SWO-38. Chin Pharm Bull 22:184–188Google Scholar
- 5.Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996. doi: 10.1016/j.canrad.2005. 07.001 Google Scholar
- 13.Peng X, Guo W, Liu T, Wang X, Tu X, Xiong D, Chen S, Lai Y, Du H, Chen G, Liu G, Tang Y, Huang S, Zou X (2011) Identification of miRs-143 and -145 that is associated with bone metastasis of prostate cancer and involved in the regulation of EMT. PLoS ONE 6:e20341. doi: 10.1371/journal.pone.0020341 PubMedCrossRefGoogle Scholar
- 14.Andersen CL, Jensen JL, Orntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250. doi: 10.1158/0008-5472.CAN-04-0496 PubMedCrossRefGoogle Scholar
- 16.Uziel T, Karginov FV, Xie S, Parker JS, Wang YD, Gajjar A, He L, Ellison D, Gilbertson RJ, Hannon G, Roussel MF (2009) The miR-17~92 cluster collaborates with the Sonic Hedgehog pathway in medulloblastoma. Proc Natl Acad Sci USA 106:2812–2817. doi: 10.1073/pnas.0809579106 PubMedCrossRefGoogle Scholar
- 17.Brabletz T (2012) MiR-34 and SNAIL: Another double-negative feedback loop controlling cellular plasticity/EMT governed by p53. Cell Cycle 11:215. doi: 10.4161/cc.11.2.18900
- 21.Mikheeva SA, Mikheev AM, Petit A, Beyer R, Oxford RG, Khorasani L, Maxwell JP, Glackin CA, Wakimoto H, González–Herrero I, Sánchez–García I, Silber JR, Horner PJ, Rostomily RC (2010) TWIST1 promotes invasion through mesenchymal change in human glioblastoma. Mol Cancer 9:194. http://www.molecular-cancer.com/content/9/1/194 Google Scholar
- 29.Zhou Y, Chen KS, Gao JB, Han R, Lu JJ, Peng T, Jia YJ (2012) miR-124-1 promotes neural differentiation of rat bone marrow mesenchymal stem cells. Zhong guo Dang Dai Er Ke Za Zhi 14: 215–220. http://www.cjcp.org/EN/Y2012/V14/I3/215