Protein lysate microarray analysis to identify microRNAs regulating estrogen receptor signaling in breast cancer cell lines

Suvi-Katri Leivonen (Corresponding Author), Rami Mäkelä, Päivi Östling, Pekka Kohonen, Saija Haapa-Paananen, Kristine Kleivi, E. Enerly, Anna Aakula, K. Hellström, Niko Sahlberg, V.N. Kristensen, A.-L. Børresen-Dale, Petri Saviranta, Merja Perälä, Olli Kallioniemi

Research output: Contribution to journalArticleScientificpeer-review

178 Citations (Scopus)

Abstract

Predicting the impact of microRNAs (miRNAs) on target proteins is challenging because of their different regulatory effects at the transcriptional and translational levels. In this study, we applied a novel protein lysate microarray (LMA) technology to systematically monitor for target protein levels after high-throughput transfections of 319 pre-miRs into breast cancer cells. We identified 21 miRNAs that downregulated the estrogen receptor-α (ERα), as validated by western blotting and quantitative real time–PCR, and by demonstrating the inhibition of estrogen-stimulated cell growth. Five potent ERα-regulating miRNAs, miR-18a, miR-18b, miR-193b, miR-206 and miR-302c, were confirmed to directly target ERα in 3′-untranslated region reporter assays. The gene expression signature that they repressed highly overlapped with that of a small interfering RNA against ERα, and across all the signatures tested, was most closely associated with the repression of known estrogen-induced genes. Furthermore, miR-18a and miR-18b showed higher levels of expression in ERα-negative as compared with ERα-positive clinical tumors. In summary, we present systematic and direct functional evidence of miRNAs inhibiting ERα signaling in breast cancer, and demonstrate the high-throughput LMA technology as a novel, powerful technique in determining the relative impact of various miRNAs on key target proteins and associated cellular processes and pathways.
Original languageEnglish
Pages (from-to)3926-3936
Number of pages11
JournalOncogene
Volume28
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint Dive into the research topics of 'Protein lysate microarray analysis to identify microRNAs regulating estrogen receptor signaling in breast cancer cell lines'. Together they form a unique fingerprint.

Cite this