n-caproylsphingosine and Prostatic-Neoplasms

The nomenclature for the serine/threonine protein phosphatases was established by Professor Sir Philip Cohen over 30 years ago. (1) At that time protein phosphatase 1 was known to have two small inhibitory proteins (I-1 and I-2) and be regulated by sub-cellular location whereas no protein inhibitor had yet been discovered for the related multi-subunit phosphatase PP2A. That paradigm subsequently changed, and several PP2A protein inhibitors have been discovered. (2) The protein I2PP2A (SET) is considered to be oncogenic, i.e., PP2A is a tumor suppressor, and is overexpressed in many tumor cell types (ref. 3, and refs. therein). I2PP2A also has other targets besides PP2A, e.g., DNA exonucleases and modification of histone acetylation. (4) PP2A activity is known to be regulated by the bioactive lipid ceramide, and this occurs through both I2PP2A inhibition and PP2A de-repression and through ceramide actions on subunits of the PP2A enzyme complex. (5)(,) (6) In the present manuscript the authors examined the expression of I2PP2A in prostate cancer and prostate epithelial cells. They determined whether ceramide could decrease accumulation of the oncogene c-Myc through inhibition of I2PP2A and activation of PP2A. As I2PP2A is also an inhibitor of histone acetylation they determined whether ceramide could block the epigenetic action of I2PP2A.

Topics: Antineoplastic Agents; Ceramides; DNA-Binding Proteins; Histone Chaperones; Humans; Male; Prostatic Neoplasms; Transcription Factors

Inhibitor 2 of protein phosphatase 2A (I2PP2A), a biological inhibitor of the cellular serine/threonine protein phosphatase PP2A, is associated with numerous cellular processes that often lead to the formation and progression of cancer. In this study we hypothesized that targeting the inhibition of I2PP2A's multiple functions in prostate cancer cells might prevent cancer progression. We have investigated the effect of the small chain C6-ceramide, known to be a bioactive tumor suppressor lipid, on I2PP2A function, thereby affecting c-Myc signaling and histone acetylation in cells. Our data indicated that C6-ceramide treatment of prostate cancer cells induces cell death in PC-3, DU145, and LNCaP cells, but not normal prostate epithelial cells. C6-ceramide was able to disrupt the association between PP2A and I2PP2A. C6-ceramide inhibits I2PP2A's upregulation of c-Myc and downregulation of histone acetylation in prostate cancer cells. Our data indicated that targeting cancer related signaling pathways through I2PP2A using ceramide as an anti-I2PP2A agent could have beneficial effects as a therapeutic approach to prevent prostate cancer.

Topics: Acetylation; Antineoplastic Agents; Cell Line, Tumor; Cell Survival; Ceramides; DNA-Binding Proteins; Epithelial Cells; Gene Knockdown Techniques; Histone Chaperones; Histones; Humans; Male; Molecular Targeted Therapy; Phosphoprotein Phosphatases; Prostatic Neoplasms; Protein Phosphatase 2C; Protein Processing, Post-Translational; RNA, Small Interfering; Transcription Factors

Stress-induced activation and metabolism of plasma membrane sphingolipids results in intracellular ceramide accumulation and has been shown to induce apoptosis in human prostate cancer cells. This effect has been observed using synthetic ceramide analogs, such as C6-ceramide; however, the effects of naturally-occurring sphingolipids, such as C18-ceramide and sphingomyelin (CerPCho), on apoptosis and prostate cancer cell proliferation have not been examined. The results of the present study demonstrate that natural (CerPCho, C18-ceramide) and synthetic (C6-ceramide) sphingolipids reduced PC-3 cell proliferation by 15 +/- 1.8, 17 +/- 2.5, and 46 +/- 2.1%, respectively (P < 0.05). These reductions in proliferation were due, in part, to increased cellular apoptosis. Treatment of PC-3 cells with CerPCho and C18-ceramide significantly increased apoptosis by 3.0 +/- 0.8 and 3.6 +/- 0.6%, respectively, compared to the untreated control, while the synthetic C6-ceramide significantly increased apoptosis by 55.7 +/- 0.4%. C6-ceramide-induced apoptosis was associated with cell cycle arrest in the G(2)/M phase, decreased extracellular signal-regulated kinase (ERK1/2) signaling and activation of the cell cycle regulatory protein, retinoblastoma (pRb). Treatment of PC-3 cells with C18-ceramide and CerPCho did not alter cell cycle distribution, pRb or ERK1/2 activation. Taken together, these results suggest that natural and synthetic sphingolipids induce apoptosis in PC-3 cells via distinct signaling mechanisms and potencies.

Topics: Androgens; Apoptosis; Cell Cycle Proteins; Cell Proliferation; Ceramides; Diet; Humans; Male; Mitogen-Activated Protein Kinase 3; Prostatic Neoplasms; Sphingolipids; Tumor Cells, Cultured

Among the many processes regulating cell death, ceramide signaling is a vital component. We previously determined that acid ceramidase (AC) is upregulated in 60% of primary prostate cancer (PCa) tissues, suggesting that AC may play a role in tumor development. In order to determine the significance of AC elevation, stable clones of DU145 cells with AC overexpression (AC-EGFP) were generated. Compared to controls (EGFP), AC-EGFP cells exhibited enhanced cell proliferation and migration. Subcutaneous injection of AC-EGFP cells into Nu/Nu mice resulted in larger tumor volumes compared to EGFP controls. Moreover, using the MTS viability assay, AC-EGFP cells were more resistant to cell death induced by doxorubicin, cisplatin, etoposide, gemcitabine or C6-ceramide. Conversely, knock down of AC using siRNA, sensitized AC-EGFP cells to these drugs. In addition, mass spectroscopic analysis of sphingolipids indicated that long chain ceramide levels were decreased in AC-EGFP cells treated with either doxorubicin or etoposide. In conclusion, this study implicates AC as a critical regulator of PCa progression by affecting not only tumor cell proliferation and migration but also responses to drug therapy, suggesting AC as a potential therapeutic target in advanced PCa.

Topics: Animals; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Ceramides; Disease Progression; Drug Resistance, Neoplasm; Galactosylgalactosylglucosylceramidase; Gene Expression Regulation, Neoplastic; Humans; Male; Mice; Mice, Nude; Prostatic Neoplasms