Vector-Based let-7a miRNAs Function as Suppressors of Multiple Oncogenes in SK-N-MC and SHEP Neuroblastoma Cells
- *Corresponding Author:
- Guan J
Department of Scientific Research
Department of Pathology, PUMC Hospital
CAMS/PUMC, No.1, Shuaifuyuan Street, Dongcheng
District, Beijing 100730 China
E-mail: [email protected]
Received date: September 30, 2015; Accepted date: January 19, 2015; Published date: January 22, 2015
Citation: Guan J, Guo S, Zeng X, Luo Y, Yang T, et al. (2016) Vector-Based let-7a miRNAs Function as Suppressors of Multiple Oncogenes in SK-N-MC and SHEP Neuroblastoma Cells. J Carcinog Mutagene 7:248. doi: 10.4172/2157-2518.1000248
Copyright: © 2015, Guan J. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Objective: This study was designed to determine the anti-tumor effect of vector-based let-7a miRNA in neuroblastoma cell lines.
Methods: A tetracycline (tet)-inducible let-7a expression vector was constructed and used to stably transfect SKN- MC and SHEP neuroblastoma cells. The effects of let-7a overexpression induced by tetracycline on cell proliferation and adhesion were analyzed in vitro by MTT assays and adhesion assays. The mRNA expression of the let-7 target oncogenes NRAS, KRAS, c-myc, and HMGA2 was examined by quantitative real-time reverse transcription polymerase chain reaction. Protein expression of N-RAS, K-RAS, c-Myc, HMGA2, NeuN, and β3- tubulin was analyzed using western blot and immunocytochemistry. Morphology of SK-N-MC and SHEP cells was also observed. Additionally, the let-7a-inducible vector-transfected SHEP cells were subcutaneously injected in nude mice, and tumor growth in vivo was also evaluated.
Results: Let-7a expression was significantly induced in neuroblastoma cells and inhibited cell proliferation and adhesion in both SK-N-MC and SHEP cells. Let-7a up-regulated β3-tubulin and NeuN expression in SK-N-MC and SHEP cells. However, short neurite-like outgrowth was observed only in SK-N-MC cells, but not in SHEP cells. Let-7a overexpression decreased the expression of N-RAS and HMGA2 proteins in both SK-N-MC and SHEP cells. c-Myc expression also decreased in SK-N-MC cells. The tumor sizes of SHEP xenografts in the tet-inducible group were smaller than those in the mice without tetracycline induction.
Conclusion: Taken together, our data demonstrate the anti-tumor effects of vector-based let-7a miRNA overexpression in SK-N-MC and SHEP neuroblastoma cells against multioncogenes. Let-7a miRNA is a potential therapeutic molecule for the treatment of neuroblastoma.