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Ectopic expression of miR-126*, an intronic product of the vascular endothelial EGF-like 7 gene, regulates prostein translation and invasiveness of prostate cancer LNCaP cells

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Abstract

MicroRNAs (miRNAs) are endogenous noncoding RNAs that down-regulate gene expression by promoting cleavage or translational arrest of target mRNAs. While most miRNAs are transcribed from their own dedicated genes, some map to introns of ‘host’ transcripts, the biological significance of which remains unknown. Here, we show that prostate cells are naturally devoid of EGF-like domain 7 (Egfl7) transcripts and hence also deficient in a miRNA, miR-126*, generated from splicing and processing of its ninth intron. Use of recombinant and synthetic miRNAs or a specific antagomir established a role of miR-126* in silencing prostein in non-endothelial cells. We mapped two miR-126*-binding sites in the 3′UTR of the prostein mRNA required for translational repression. Transfection of synthetic miR-126* into prostate cancer LNCaP cells strongly reduced the translation of prostein. Interestingly, loss of prostein correlated with reduction of LNCaP cell migration and invasion. Thus, the robust expression of prostein protein in the prostate cells results from a combination of transcriptional activation of the prostein gene and absence of intronic miRNA-126* due to the prostate-specific repression of the Egfl7 gene. We conclude that intronic miRNAs from tissue-specific transcripts, or their natural absence, make cardinal contributions to cellular gene expression and phenotype. These findings also open the door to tissue-specific miRNA therapy.

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Abbreviations

bp:

base pair

RNAi:

RNA interference

siRNA:

small interfering RNA

miRNA:

microRNA

RISC:

RNAi-induced silencing complex

RSV:

respiratory syncytial virus

UTR:

untranslated region

(Egfl7):

EGF-like domain 7

GAPDH:

glyceraldehyde 3-phosphate dehydrogenase

HUVEC:

human umbilical vein endothelial cells

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Acknowledgments

This work was supported in part by the US National Institute of Health (NIH) grant AI059267 to S.B. We thank Dr. Ratna Chakrabarti (University of Central Florida) for the LNCaP cells, Dr. Lalita Shevde-Samant (Mitchell Cancer Institute) for the PC3 and DU145 cell lines and advice with the Boyden chamber and wounding assays, Dr. Ramesh Pillai (EMBL Grenoble, France) for the pRL-Con plasmid, and Dr. Lance Ford (Bioo Scientific Corp., Austin, Texas) for helpful discussions on miRNA analysis and assay.

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  1. Department of Biochemistry and Molecular Biology, University of South Alabama, College of Medicine, 307 University Blvd., Mobile, Alabama, 36688-0002, USA

    Alla Musiyenko, Vira Bitko & Sailen Barik

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  1. Alla Musiyenko
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  2. Vira Bitko
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Correspondence to Sailen Barik.

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Musiyenko, A., Bitko, V. & Barik, S. Ectopic expression of miR-126*, an intronic product of the vascular endothelial EGF-like 7 gene, regulates prostein translation and invasiveness of prostate cancer LNCaP cells. J Mol Med 86, 313–322 (2008). https://doi.org/10.1007/s00109-007-0296-9

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  • DOI: https://doi.org/10.1007/s00109-007-0296-9

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