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MicroRNAs (miRNAs) are a class of small non-coding RNAs (20 to 24 nucleotides) that post-transcriptionally modulate gene expression. A key oncomir in carcinogenesis is  miR-21 , which is consistently up-regulated in a wide range of cancers. However, few functional studies are available for  miR-21 , and few targets have been identified. In this study, we explored the role of  miR-21  in human breast cancer cells and tissues, and searched for  miR-21  targets.    Methods  We used  in vitro  and  in vivo  assays to explore the role of  miR-21  in the malignant progression of human breast cancer, using  miR-21  knockdown. Using LNA silencing combined to microarray technology and target prediction, we screened for potential targets of  miR-21  and validated direct targets by using luciferase reporter assay and Western blot. Two candidate target genes (EIF4A2 and ANKRD46) were selected for analysis of correlation with clinicopathological characteristics and prognosis using immunohistochemistry on cancer tissue microrrays.    Results  Anti-miR-21 inhibited growth and migration of MCF-7 and MDA-MB-231 cells  in vitro , and tumor growth in nude mice. Knockdown of  miR-21  significantly increased the expression of  ANKRD46  at both mRNA and protein levels. Luciferase assays using a reporter carrying a putative target site in the 3' untranslated region of  ANKRD46  revealed that  miR-21  directly targeted  ANKRD46 .  miR-21  and EIF4A2 protein were inversely expressed in breast cancers (r s  = -0.283,  P  = 0.005, Spearman's correlation analysis).    Conclusions  Knockdown of  miR-21  in MCF-7 and MDA-MB-231 cells inhibits  in vitro  and  in vivo  growth as well as  in vitro  migration.  ANKRD46  is newly identified as a direct target of  miR-21  in BC. These results suggest that inhibitory strategies against  miR-21  using peptide nucleic acids (PNAs)-antimiR-21 may provide potential therapeutic applications in breast cancer treatment.

Knockdown of miR-21 in human breast cancer cell lines inhibits proliferation, in vitro migration and in vivotumor growth