MicroRNA15a modulates expression of the cell-cycle regulator Cdc25A and affects hepatic cystogenesis in a rat model of polycystic kidney disease
Seung-Ok Lee, … , Angela Stroope, Nicholas LaRusso
Seung-Ok Lee, … , Angela Stroope, Nicholas LaRusso
Published October 23, 2008
Citation Information: J Clin Invest. 2008;118(11):3714-3724. https://doi.org/10.1172/JCI34922.
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Research Article

MicroRNA15a modulates expression of the cell-cycle regulator Cdc25A and affects hepatic cystogenesis in a rat model of polycystic kidney disease

Abstract

Hyperproliferation of bile duct epithelial cells due to cell-cycle dysregulation is a key feature of cystogenesis in polycystic liver diseases (PCLDs). Recent evidence suggests a regulatory role for microRNAs (miRNAs) in a variety of biological processes, including cell proliferation. We therefore hypothesized that miRNAs may be involved in the regulation of selected components of the cell cycle and might contribute to hepatic cystogenesis. We found that the cholangiocyte cell line PCK-CCL, which is derived from the PCK rat, a model of autosomal recessive polycystic kidney disease (ARPKD), displayed global changes in miRNA expression compared with normal rat cholangiocytes (NRCs). More specific analysis revealed decreased levels of 1 miRNA, miR15a, both in PCK-CCL cells and in liver tissue from PCK rats and patients with a PCLD. The decrease in miR15a expression was associated with upregulation of its target, the cell-cycle regulator cell division cycle 25A (Cdc25A). Overexpression of miR15a in PCK-CCL cells decreased Cdc25A levels, inhibited cell proliferation, and reduced cyst growth. In contrast, suppression of miR15a in NRCs accelerated cell proliferation, increased Cdc25A expression, and promoted cyst growth. Taken together, these results suggest that suppression of miR15a contributes to hepatic cystogenesis through dysregulation of Cdc25A.

Authors

Seung-Ok Lee, Tatyana Masyuk, Patrick Splinter, Jesús M. Banales, Anatoliy Masyuk, Angela Stroope, Nicholas LaRusso

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Figure 7

Targeting of miR15a to the 3′-UTR region of Cdc25A mRNA.

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Targeting of miR15a to the 3′-UTR region of Cdc25A mRNA. (A) 2 reporter ...
(A) 2 reporter constructs (Cdc25A WT and Cdc25A mutant) with the potential binding site for miR15a in the 3′-UTR region of Cdc25A were generated. (B) In PCK-CCL cotransfected with the Cdc25A WT construct and miR15a precursor (pre-miR15a), luciferase reporter translation was significantly decreased compared with that in PCK-CCL transfected with the Cdc25A WT construct only. A mutation in the miR15a–binding sequence in the Cdc25A mRNA 3′-UTR eliminated this effect. Moreover, in the presence of scrambled miRNA (scr miR), no changes in luciferase reporter were observed. Data are representative of 7 independent experiments. *P < 0.05.