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 8

Effect of miR15a overexpression in cystic cholangiocytes on miR15a levels, cell proliferation, and cyst growth.

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Effect of miR15a overexpression in cystic cholangiocytes on miR15a level...
(A) Quantitative PCR shows that in PCK-CCL stably transfected with the miR15a precursor (PCK-CCL–pSR-miR15a), levels of miR15a were increased compared with those in untransfected PCK-CCL or PCK-CCL transfected with EV (PCK-CCL–pSR-EV). (B) Cell proliferation was inhibited in PCK-CCL–pSR-miR15a compared with that in PCK-pSR-EV. Data are representative of 3 independent experiments. (C) Light microscopy analysis of PCK-CCL–pSR-EV and PCK-CCL–pSR-miR15a revealed the presence of cystic structures in 3D culture. Cysts formed by PCK-CCL–pSR-EV (top panels) continued to grow progressively over time while cystic structures formed by PCK-CCL–pSR-miR15a (bottom panels) did not expand. Scale bars: 250 μm. (D) Quantitative analysis of cystic areas demonstrates that rates of cyst growth in PCK-CCL–pSR-miR15a cultures (n = 13) were inhibited significantly compared with those in PCK-CCL–pSR-EV cultures (n = 25). *P < 0.05.