Abstract
Peroxisome proliferator-activated receptor-gamma (PPARγ) regulates the interface between cellular lipid metabolism, redox status and organelle differentiation. Conditional prostatic epithelial knockout of PPARγ in mice resulted in focal hyperplasia which developed into mouse prostatic intraepithelial neoplasia (mPIN). The grade of PIN became more severe with time. Electron microscopy (EM) showed accumulated secondary lysosomes containing cellular organelles and debris suggestive of autophagy. Consistent with this analysis the autophagy marker LC-3 was found to be upregulated in areas of PIN in PPARγ KO tissues. We selectively knocked down PPARγ2 isoform in wild-type mouse prostatic epithelial cells and examined the consequences of this in a tissue recombination model. Histopathologically grafted tissues resembled the conditional PPARγ KO mouse prostates. EM studies of PPARγ- and PPARγ2-deficient epithelial cells in vitro were suggestive of autophagy, consistent with the prostatic tissue analysis. This was confirmed by examining expression of beclin-1 and LC-3. Gene expression profiling in PPARγ-/γ2-deficient cells indicated a major dysregulation of cell cycle control and metabolic signaling networks related to peroxisomal and lysosomal maturation, lipid oxidation and degradation. The putative autophagic phenotypes of PPARγ-deficient cells could be rescued by re-expression of either γ1 or γ2 isoform. We conclude that disruption of PPARγ signaling results in autophagy and oxidative stress during mPIN pathogenesis.
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Abbreviations
- Pca:
-
prostate cancer or prostate carcinoma
- PIN:
-
prostatic intraepithelial neoplasia
- PPRE:
-
peroxisome proliferator response element
- mPrE:
-
a non-malignant mouse prostate epithelial cell line spontaneously immortalized from an adult wild-type C57Bl6 mouse
- mPrE-PPARγ KO (mPrE-γKO):
-
a PPARγ knockout mouse prostate epithelial cell line spontaneously immortalized from an adult PBCre4 tg/0/PPARγ flox/flox mouse prostate
- mPrE-pSIR-PPARγ2 shRNA (mPrE-γ2sh):
-
a PPARγ2 knockdown mouse prostate epithelial cell line transfected by U6-mouse PPARγ2 shRNA in pSIR retroviral vector into mPrE cells
- mPrE-pSIR-empty vector (mPrE-pSIR):
-
a control pSIR vector-transfected mouse prostate epithelial cell line
- mPrE-PPARγKO-PPARγ1 wild-type (WT) (mPrE-γKO-γ1WT):
-
mPrE-γKO overexpressing mouse PPARγ1 WT full-length cDNA
- mPrE-PPARγKO-PPARγ2 wild-type (WT) (mPrE-γKO-γ2WT):
-
mPrE-γKO overexpressing mouse PPARγ2 WT full-length cDNA
- mPrE-PPARγKO-empty vector (EV) (mPrE-γKO-EV) transduction control:
-
mPrE-γKO retrovirally transduced using a control empty vector
- UGS:
-
urogenital sinus
- UGM:
-
urogenital mesenchymal cell
- KO:
-
knockout
- KD:
-
knockdown
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Acknowledgements
The authors appreciate EM core lab members for help with E.M., Vanderbilt MCN III Animal facility for animal care, Vanderbilt Microarray Shared Resource (VMSR) facility for microarray work. The pCMX-mouse PPARγ1 and PPARγ2 wild-type full-length cDNA expression plasmids and PPARγ-Luciferase reporter plasmid (PPRE)3-tk-luciferase were gifts from Dr. V.K.K. Chatterjee, University of Cambridge, Cambridge, UK. The pQCXIP-mouse PPARγ1 and PPARγ2 wild-type full-length cDNA vectors were gifts from Drs. Y. Eugene Chen and Jifeng Zhang, University of Michigan Medical Center. The CK-14 (LL001) antibody was a gift from Dr. EB Lane, Universty of Dundee, Scotland and the α-LC-3 antiserum from Dr. Tamotsu Yoshimori, National Institute of Genetics (NIG), Mishima, Japan. The work was supported by NIH R01 DK67049 and DOD-PCRP, W81XWH-07-1-0479 grants to S.W.H. NIH R01 CA113392 and NIH R01 CA59705 grants to P.R-B, NIH R01 DK055748 to R.J.M. The VUMC Research EM Resource is supported by NIH grants DK20539, CA68485 and DK58404. The Vanderbilt Microarray Shared Resource is supported by the Vanderbilt Ingram Cancer Center (P30 CA68485), the Vanderbilt Digestive Disease Center (P30 DK58404) and the Vanderbilt Vision Center (P30 EY08126). We also thank the Frances Williams Preston Laboratories of the TJ Martell Foundation for support. The content of this paper is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Jiang, M., Fernandez, S., Jerome, W. et al. Disruption of PPARγ signaling results in mouse prostatic intraepithelial neoplasia involving active autophagy. Cell Death Differ 17, 469–481 (2010). https://doi.org/10.1038/cdd.2009.148
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DOI: https://doi.org/10.1038/cdd.2009.148
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