Methods to Quantify MicroRNAs in the Myc Gene Network for Posttranscriptional Gene Repression

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2013 Sep 6

PMID 24006063

Citations 2

Authors

Rui Song

Nicole Sponer

Lin He

Affiliations

Soon will be listed here.

Abstract

As a global transcription factor, Myc regulates both protein-coding genes and noncoding microRNA genes. Myc-activated or repressed miRNAs are involved in various pathways to affect tumorigenesis, mediate apoptosis, proliferation, angiogenesis, metastasis, and metabolism downstream of Myc. Functional characterization of miRNAs in the Myc network requires the accurate detection and quantification of miRNA expression levels. Here, we describe two widely used methodologies to determine miRNA expression, including miRNA real-time PCR and miRNA northern analysis.

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References

Hong L, Lai M, Chen M, Xie C, Liao R, Kang Y . The miR-17-92 cluster of microRNAs confers tumorigenicity by inhibiting oncogene-induced senescence. Cancer Res. 2010; 70(21):8547-57. PMC: 2970743. DOI: 10.1158/0008-5472.CAN-10-1938. View

Cimmino A, Calin G, Fabbri M, Iorio M, Ferracin M, Shimizu M . miR-15 and miR-16 induce apoptosis by targeting BCL2. Proc Natl Acad Sci U S A. 2005; 102(39):13944-9. PMC: 1236577. DOI: 10.1073/pnas.0506654102. View

Pall G, Hamilton A . Improved northern blot method for enhanced detection of small RNA. Nat Protoc. 2008; 3(6):1077-84. DOI: 10.1038/nprot.2008.67. View

Meyer N, Penn L . Reflecting on 25 years with MYC. Nat Rev Cancer. 2008; 8(12):976-90. DOI: 10.1038/nrc2231. View

Chou Y, Lin H, Lien Y, Wang Y, Hong C, Kao Y . EGFR promotes lung tumorigenesis by activating miR-7 through a Ras/ERK/Myc pathway that targets the Ets2 transcriptional repressor ERF. Cancer Res. 2010; 70(21):8822-31. DOI: 10.1158/0008-5472.CAN-10-0638. View

Tazawa H, Tsuchiya N, Izumiya M, Nakagama H . Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells. Proc Natl Acad Sci U S A. 2007; 104(39):15472-7. PMC: 2000550. DOI: 10.1073/pnas.0707351104. View

He L, Hannon G . MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet. 2004; 5(7):522-31. DOI: 10.1038/nrg1379. View

Ma L, Young J, Prabhala H, Pan E, Mestdagh P, Muth D . miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol. 2010; 12(3):247-56. PMC: 2845545. DOI: 10.1038/ncb2024. View

Kota J, Chivukula R, ODonnell K, Wentzel E, Montgomery C, Hwang H . Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model. Cell. 2009; 137(6):1005-17. PMC: 2722880. DOI: 10.1016/j.cell.2009.04.021. View

10.

He L, He X, Lim L, de Stanchina E, Xuan Z, Liang Y . A microRNA component of the p53 tumour suppressor network. Nature. 2007; 447(7148):1130-4. PMC: 4590999. DOI: 10.1038/nature05939. View

11.

Mu P, Han Y, Betel D, Yao E, Squatrito M, Ogrodowski P . Genetic dissection of the miR-17~92 cluster of microRNAs in Myc-induced B-cell lymphomas. Genes Dev. 2009; 23(24):2806-11. PMC: 2800095. DOI: 10.1101/gad.1872909. View

12.

Gao P, Tchernyshyov I, Chang T, Lee Y, Kita K, Ochi T . c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature. 2009; 458(7239):762-5. PMC: 2729443. DOI: 10.1038/nature07823. View

13.

He L, Thomson J, Hemann M, Hernando-Monge E, Mu D, Goodson S . A microRNA polycistron as a potential human oncogene. Nature. 2005; 435(7043):828-33. PMC: 4599349. DOI: 10.1038/nature03552. View

14.

Ro S, Park C, Jin J, Sanders K, Yan W . A PCR-based method for detection and quantification of small RNAs. Biochem Biophys Res Commun. 2006; 351(3):756-63. PMC: 1934510. DOI: 10.1016/j.bbrc.2006.10.105. View

15.

Dews M, Homayouni A, Yu D, Murphy D, Sevignani C, Wentzel E . Augmentation of tumor angiogenesis by a Myc-activated microRNA cluster. Nat Genet. 2006; 38(9):1060-5. PMC: 2669546. DOI: 10.1038/ng1855. View

16.

Chang T, Wentzel E, Kent O, Ramachandran K, Mullendore M, Lee K . Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. Mol Cell. 2007; 26(5):745-52. PMC: 1939978. DOI: 10.1016/j.molcel.2007.05.010. View

17.

Johnson C, Esquela-Kerscher A, Stefani G, Byrom M, Kelnar K, Ovcharenko D . The let-7 microRNA represses cell proliferation pathways in human cells. Cancer Res. 2007; 67(16):7713-22. DOI: 10.1158/0008-5472.CAN-07-1083. View

18.

Raver-Shapira N, Marciano E, Meiri E, Spector Y, Rosenfeld N, Moskovits N . Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. Mol Cell. 2007; 26(5):731-43. DOI: 10.1016/j.molcel.2007.05.017. View

19.

ODonnell K, Wentzel E, Zeller K, Dang C, Mendell J . c-Myc-regulated microRNAs modulate E2F1 expression. Nature. 2005; 435(7043):839-43. DOI: 10.1038/nature03677. View

20.

Dang C . MYC on the path to cancer. Cell. 2012; 149(1):22-35. PMC: 3345192. DOI: 10.1016/j.cell.2012.03.003. View