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Tissue-Specific Stem Cells
Version of Record online: 20 AUG 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 9, pages 1985–1998, September 2012
How to Cite
Fonsato, V., Collino, F., Herrera, M. B., Cavallari, C., Deregibus, M. C., Cisterna, B., Bruno, S., Romagnoli, R., Salizzoni, M., Tetta, C. and Camussi, G. (2012), Human Liver Stem Cell-Derived Microvesicles Inhibit Hepatoma Growth in SCID Mice by Delivering Antitumor MicroRNAs. STEM CELLS, 30: 1985–1998. doi: 10.1002/stem.1161
Author contributions: V.F. and F.C.: experiment design, collection and assembly of data, data analysis and interpretation, and manuscript writing; M.B.H., M.C.D., S.B., and C.C.: collection and/or assembly of data and data analysis and interpretation; B.C.: collection and/or assembly of data; R.R. and M.S.: provision of study material and data analysis and interpretation; C.T.: data analysis and interpretation, financial support, and manuscript writing; G.C.: conception and design, financial support, data analysis and interpretation, and manuscript writing. V.F. and F.C. contributed equally to this work.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS June 26, 2012.
Re-use of this article is permitted in accordance with the Terms and Conditions set out at http://wileyonlinelibrary.com/onlineopen#OnlineOpen_Terms
- Issue online: 20 AUG 2012
- Version of Record online: 20 AUG 2012
- Accepted manuscript online: 26 JUN 2012 03:43PM EST
- Manuscript Accepted: 5 JUN 2012
- Manuscript Received: 17 FEB 2012
- Associazione Italiana per la Ricerca sul Cancro (A.I.R.C.). Grant Number: IG8912
- Italian Ministry of University and Research (MIUR) Prin08
- Regione Piemonte, Project Oncoprot, and Piattaforme Biotecnologiche, Pi-Stem project
Additional Supporting Information may be found in the online version of this article.
|SC_12-0166_sm_supplFigure1.tif||4135K||Supplemental Figure 1. (A) qRT-PCR analysis (left panel) of fibroblasts transfected with pCMV-Sport 6 plasmid carrying the human CD29 (fibr-CD29) full length cDNA or of fibroblasts electroporated in the absence of DNA as control (fibr-CTR). Data were normalized to 1 for fibr-CTR and to GAPDH mRNA. * P < 0.01 vs fibr-CTR. Right panel, Western Blot analysis of CD29 in fibr-CTR, in MVs derived from fibr-CTR, in fibr-CD29 and in MVs derived from fibr-CD29. CD29 transfected fibroblasts released MVs expressing CD29. (B) Nanoparticle tracking analysis (NTA) showing that the RNase treatment (right panel) did not affect MV morphology and size. Left panel shows untreated MV-HLSC. Curve 1 describes the relationship between particle number distribution (left Y axis) and particle size (X axis); curve 2 shows cumulative percentage distribution of particles (percentile in right Y axis) and particle size (X axis). Mean size and particle concentration values were calculated by the NTA software that allow analysis of video images of the particle movement under Brownian motion captured by Nanosight LM10 and calculation of the diffusion coefficient, sphere equivalent, and hydrodynamic radius of particles by using the Strokes-Einstein equation. (C) Measure of selected miRNAs in MVHLSC treated 5U/ml (white bar) of RNase in respect to control MV-HLSC (black bar) expressed as 2-(ΔCt) as described in Methods. Data were normalized to 1 for MV-HLSC and expressed as mean ± SD of 4 different experiments. * P < 0.01 vs MV-HLSC. (D) Measure in MV-HLSC of the levels of different miRNAs relative to the small nucleolar RNA, RNU48 expressed as Log 2-(ΔCt).|
|SC_12-0166_sm_supplFigure2.pdf||911K||Supplemental Figure 2. Evaluation of in vivo tumor proliferation and apoptosis (A) Representative micrographs of PCNA staining of HepG2 tumors. Scale bar, 50 μm. (B) Representative micrographs of Tunel staining. Scale bar, 50 μm.|
|SC_12-0166_sm_supplFigure3.tif||344K||Supplemental Figure 3. Comparative expression of specific miRNAs and correspondent targets between HepG2 and normal human hepatocytes. (A) Log of relative expression of miR451, miR223, miR31 and miR21 in normal human hepatocytes (Hep; white bar) in respect to HepG2, evaluated by qRT-PCR. (B) qRTPCR analysis of miR451 and miR31 correspondent targets in HepG2 (grey bar) and in Hep (white bar). Data (A and B) were normalized to 18S ribosomal 1 mRNA and to 1 for HepG2 and expressed as mean ± SD of 4 different experiments. * P < 0.05 vs HepG2.|
|SC_12-0166_sm_supplTable1.pdf||60K||Supplementary Table 1|
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