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 November 3, 2008; First 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 1

Rates of cell proliferation and mitotic indices in NRCs and PCK-CCL.

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Rates of cell proliferation and mitotic indices in NRCs and PCK-CCL. (A)...
(A) In PCK-CCL, the rate of cell proliferation (determined by MTS absorbance at 490 nm) was significantly higher at each time point. Data are representative of 5 independent experiments. (B) Mitotic figures (asterisks) in NRCs and PCK-CCL were assessed by immunofluorescent confocal microscopy using an anti–α-tubulin antibody as a marker for microtubules (red). Nuclei were stained with DAPI (blue). Original magnification, ×60. (C) Quantitative analysis shows that in PCK-CCL, the number of mitotic cells (>500 cells counted in each experiment) was significantly greater compared with NRCs. *P < 0.05.