These authors contributed equally to this work.
MicroRNA-494 within an oncogenic microRNA megacluster regulates G1/S transition in liver tumorigenesis through suppression of mutated in colorectal cancer
Version of Record online: 22 NOV 2013
© 2013 Authors. Hepatology published by Wiley on behalf of the American Association for the Study of Liver Diseases
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
Volume 59, Issue 1, pages 202–215, January 2014
How to Cite
Lim, L., Balakrishnan, A., Huskey, N., Jones, K. D., Jodari, M., Ng, R., Song, G., Riordan, J., Anderton, B., Cheung, S.-T., Willenbring, H., Dupuy, A., Chen, X., Brown, D., Chang, A. N. and Goga, A. (2014), MicroRNA-494 within an oncogenic microRNA megacluster regulates G1/S transition in liver tumorigenesis through suppression of mutated in colorectal cancer. Hepatology, 59: 202–215. doi: 10.1002/hep.26662
Potential conflict of interest: Nothing to report.
The work was supported by fellowship supported by A*STAR (to L.L.) and by the Susan G. Komen Foundation (to A.G.), the National Institutes of Health (CA136717 and CA170447; to A.G.), the University of California San Francisco (UCSF) Program for Breakthrough Biological Research and a V-Foundation award (to A.G.), and the UCSF Liver Center (P30 DK026743).
- Issue online: 20 DEC 2013
- Version of Record online: 22 NOV 2013
- Accepted manuscript online: 2 AUG 2013 06:57AM EST
- Manuscript Accepted: 26 JUL 2013
- Manuscript Received: 9 JAN 2013
Additional Supporting Information may be found in the online version of this article.
|hep26662-sup-0001-suppfig1.tif||1097K||Supporting Information Figure 1. Doxycycline-regulated expression of MYC or/and RAS oncogenes give rise to distinct liver tumors. A. Western blot confirming the expression of MYC and RAS in oncogene driven liver tumors. B. Table summarizing the histological characteristics of tumors driven by respective oncogenes.|
|hep26662-sup-0002-suppfig2.tif||402K||Supporting Information Figure 2. Genomic organization of the Dlk1-Dio3 locus showing location of representative miRNAs|
|hep26662-sup-0003-suppfig3.tif||1009K||Supporting Information Figure 3. A. Schematic of miRNA subcluster cloning from the Dlk1-Dio3 region. B. Western blot showing expression of MYC and RAS in LT2M and LT2MR cell lines. C. Verification of miRNA subscluster expression in LT2MR stable cell lines. qRT-PCR was performed to detect expression of representative miRNAs.|
|hep26662-sup-0004-suppfig4.tif||324K||Supporting Information Figure 4. Verification of miR-494 and 495 expression in mouse liver tumors. qRT-PCR was performed to detect expression of respective miRNAs.|
|hep26662-sup-0005-suppfig5.tif||329K||Supporting Information Figure 5. Quantification of miR-494 levels in engineered LT2MR cell lines by qRT-PCR.|
|hep26662-sup-0006-suppfig6.tif||553K||Supporting Information Figure 6. A. Quantification of miR-494 expression in Hepa1-6 cell lines transfected with either a control or miR-494 antimiR, by qRT-PCR. B. Proliferation assay performed on Hepa1-6 cells transfected with either control or miR-494 antimiR. C. Cell cycle analysis following staining for DNA content performed on Hepa1-6 cells transfected with either control or miR-494 antimiR. D. Western blot analysis of MCC, p27 and p21 expression of Hepa1-6 cells cells transfected with either control or miR-494 antimiR.|
|hep26662-sup-0007-suppfig7.tif||355K||Supporting Information Figure 7. Luciferase reporter assay on TACC2, TGFB2, RB1, RB1CC1, CDC73 and WEE1 3′ UTRs.|
|hep26662-sup-0008-suppfig8.tif||3166K||Supporting Information Figure 8. A. Quantification of miR-494 levels in Huh7 and Hep3B cells relative to normal liver tissue. B. Verification of miR-494 expression in tumors treated with either control or miR-494 antimiR. miR-451 expression was tested as a negative control. qRT-PCR was performed to detect expression of respective miRNAs. C. Quantification of TUNEL staining performed on control and miR-494 antimiR treated animals. Representative pictures are shown.|
|hep26662-sup-0009-supptable1.doc||547K||Supporting Information Table 1. Differential expression of miRNAs (fold change -1 > log2 >1 and p<0.05) sorted by tumor specific expression in transgenic mouse tumor models driven by MYC, RAS or MYC+RAS. miRNAs that are upregulated, downregulated and a part of the 12q cluster are listed in separate worksheets for all three genotypes.|
|hep26662-sup-0010-supptable2.docx||36K||Supporting Information Table 2. Increased expression of miR-494 and miR-495 in transgenic mouse liver tumors. Values are expressed as fold difference compared to normal LT2 liver tissue.|
|hep26662-sup-0011-supptable3.doc||13K||Supporting Information Table 3. Primer sequences used for cloning of 12q subcluster miRNAs and 3′UTR reporter constructs.|
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