RNA-driven JAZF1-SUZ12 gene fusion in human endometrial stromal cells

Oncogenic fusion genes as the result of chromosomal rearrangements are important for understanding genome instability in cancer cells and developing useful cancer therapies. To date, the mechanisms that create such oncogenic fusion genes are poorly understood. Previously we reported an unappreciated RNA-driven mechanism in human prostate cells in which the expression of chimeric RNA induces specified gene fusions in a sequence-dependent manner. One fundamental question yet to be addressed is whether such RNA-driven gene fusion mechanism is generalizable, or rather, a special case restricted to prostate cells. In this report, we demonstrated that the expression of designed chimeric RNAs in human endometrial stromal cells leads to the formation of JAZF1-SUZ12, a cancer fusion gene commonly found in low-grade endometrial stromal sarcomas. The process is specified by the sequence of chimeric RNA involved and inhibited by estrogen or progesterone. Furthermore, it is the antisense rather than sense chimeric RNAs that effectively drive JAZF1-SUZ12 gene fusion. The induced fusion gene is validated both at the RNA and the genomic DNA level. The ability of designed chimeric RNAs to drive and recapitulate the formation of JAZF1-SUZ12 gene fusion in endometrial cells represents another independent case of RNA-driven gene fusion, suggesting that RNA-driven genomic recombination is a permissible mechanism in mammalian cells. The results could have fundamental implications in the role of RNA in genome stability, and provide important insight in early disease mechanisms related to the formation of cancer fusion genes.

Action of the natural compound esculetin on Ca2+ movement and survival in prostate cancer cells

Esculetin is derived from coumarin and is shown to be the main constituent of the Chinese herb Cortex Fraxini. The molecular paths underlying the action of esculetin are intensively studied. The outcome of esculetin on Ca2+ concentration ([Ca2+]i) in prostate cells is unexplored. Fura-2 was used to detect Ca2+ changes. Death was assessed by using WST-1. At doses of 25-100 mM, esculetin evoked [Ca2+]i raises. This signal was lessened by 15% by exclusion of Ca2+. Esculetin (100 μM) induced Mn2+ entry that implied Ca2+ influx. Esculetin-evoked Ca2+ influx was curbed by 50% by nifedipine (1 mM), econazole (0.5 mM) and SKF96365 (5 mM); phorbol 12-myristate 13 acetate (PMA; 1 nM; a protein kinase C [PKC] activator); and GF109203X (2 mM; a PKC inhibitor. In the absence of Ca2+, pretreatment with the endoplasmic reticulum (ER) Ca2+ pump inhibitor thapsigargin (1 mM) eradicated esculetin-induced [Ca2+]i raises. U73122, a phospholipase C (PLC) suppressor got rid of esculetin-caused [Ca2+]i rises. Esculetin (20-70 mM) evoked death which was not restrained by treatment with the Ca2+ binder BAPTA/AM. In summary, in PC3 cells, esculetin stimulated [Ca2+]i raises by Ca2+ influx through PKC-sensitive store-operated Ca2+ entry and PLC-associated ER Ca2+ discharging. Esculetin provoked Ca2+-independent cell death.

In Vitro Analysis of Deoxynivalenol Influence on Steroidogenesis in Prostate

Deoxynivalenol (DON) is a type-B trichothecene mycotoxin produced by Fusarium species, reported to be the most common mycotoxin present in food and feed products. DON is known to affect the production of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) in male rats, consequently affecting reproductive endpoints. Our previous study showed that DON induces oxidative stress in prostate cancer (PCa) cells, however the effect of DON on the intratumor steroidogenesis in PCa and normal prostate cells was not investigated. In this study human normal (PNT1A) and prostate cancer cell lines with different hormonal sensitivity (PC-3, DU-145, LNCaP) were exposed to DON treatment alone or in combination with dehydroepiandrosterone (DHEA) for 48 h. The results of the study demonstrated that exposure to DON alone or in combination with DHEA had a stimulatory effect on the release of estradiol and testosterone and also affected progesterone secretion. Moreover, significant changes were observed in the expression of genes related to steroidogenesis. Taken together, these results indicate that DON might affect the process of steroidogenesis in the prostate, demonstrating potential reproductive effects in humans.

Experimental evaluation of the influence of isobornylphenols on the development of BPH and the redox potential of prostate cells

Introduction. Benign prostatic hyperplasia (BPH) is a common urological disorder in older men. It is characterized by the development of glandularstromal hyperplasia of the prostate with the formation of new glandular structures and subsequent symptoms from the lower urinary tract. It has now been established that the pathogenesis of this disease is multifactorial and one of the possible mechanisms for the development of BPH is oxidative stress. Purpose. Study of the effect of phenols with a bulky isobornyl substituent (2,6-diisobornyl-4-methylphenol and 4-hydroxymethyl-2,6-diisobornylphenol)on the growth of experimental BPH and the antioxidant balance of prostate cells in comparison with Prostamol Uno. Materials and мethods. Experiments were carried out on 50 male Wistar rats. BPH was caused by daily administration of sulpiride (60 days) to male rats of late reproductive age. After 2 months, the animals were weighed and sacrificed in a CO2 chamber. The mass, mass coefficient, volume of the lateral lobe of the pancreas were determined, morphological analysis was performed. Investigated prooxidant and antioxidant activity. The results were processed by the method of variation statistics using the Mann-Whitney nonparametric U test. Results. The efficacy of the investigated drugs in BPH decreased in the following sequence: sulpiride + substance 4-hydroxymethyl-2,6-diisobornylphenol (HDB) → sulpiride + substance Dibornol (DB) → sulpiride + Prostamol Uno (PU). When comparing the results of evaluating the anti-prooxidant status with the therapeutic effect of the studied drugs, it was found that isobornylphenols, which are highly effective as prostatotropic drugs, did not show a more significant effect, compared to PU, on the redox potential of prostatic tissue cells. Conclusions. Drugs DB, HDB, PU have a normalizing effect on the level of severity of redox reactions in the sulpiride model of BPH.

Molecular and Cellular Reports A Pharmacological Study of the Action of Esculetin on Human Prostate Cancer Cells

Esculetin is a derivative of coumarin, and is the dominant, vigorous component of the conventional Chinese medicine Cortex Fraxini. Recently, the molecular pathway study and clinical use of Cortex Fraxini and esculetin are becoming intensive. In vitro, esculetin has been shown to provoke apoptotic responses via mitochondrial routes and other cellular responses in diverse cell types. The action of esculetin on cytosolic Ca2+ concentration ([Ca2+]i) in prostate cells is unknown. [Ca2+]i were assayed by applying fura-2, a fluorescent Ca2+-sensitive probe. WST-1 was used to measure cell death. Esculetin at doses of 25–100 µM provoked [Ca2+]i raises. Removing external Ca2+ decreased the response by 15%. Esculetin (100 µM) provoked Mn2+ entry implying Ca2+ influx. Esculetin-provoked Ca2+ influx was suppressed by half by protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate, PMA) and inhibitor (GF109203X); and by three inhibitors of store-operated Ca2+ channels: nifedipine, econazole and SKF96365. In the absence of Ca2+, pretreatment with the endoplasmic reticulum (ER) Ca2+ pump suppressor thapsigargin completely suppressed esculetin-provoked [Ca2+]i raises. Suppression of phospholipase C (PLC) with U73122 eliminated esculetin-provoked [Ca2+]i raises. Esculetin at 20–70 µM caused death of cells, which was not prevented by incubation with the Ca2+ binder 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). In sum, in PC3 prostate cells, esculetin provoked [Ca2+]i raises by provoking PLC-associated Ca2+ discharge from ER and Ca2+ influx via PKC-sensitive store-operated Ca2+ influx. Additionally, esculetin provoked Ca2+-dissociated cell death.

Bisphenol S Promotes the Progression of Prostate Cancer by Regulating the Expression of COL1A1 and COL1A2.

In recent decades, Bisphenol S (BPS), which have been considered as alternatives for Bisphenol A (BPA), have become widely used in personal care products, paper products and food. Clarifying the relationship between bisphenol and tumors is of great significance for the treatment and prevention of diseases. In this work, we discovered a new method to predict the correlation between bisphenol interactive genes and tumors. The transcriptome profile and interactive genes of bisphenol were obtained from the Cancer Genome Atlas and Genotype-Tissue Expression, Comparative Toxicology Genomics and PharmMapper database. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis showed that interactive genes were mainly enriched in prostate cancer. Gene targetd prediction and gene set variation analysis proved that bisphenol exert potential effects on prostate cancer. The operating characteristic curves and survival analysis showed that role of COL1A1 and COL1A2 in predicting the prognosis of prostate cancer. Cell counting kit-8 assay demonstrated that the cells with BPS-treated could remarkably promote the cell proliferation ability in both PC-3 and LNCap cells. Wound healing assay and the transwell assay demonstrated that the cells with BPS-treated could significantly promote the cell invasion capacity of prostate cells. Two key genes, COL1A1 and COL1A2, were significantly upregulated with BPS-treated in the PC-3 and LNCap cells.

Ras/ERK and PI3K/AKT signaling differentially regulate oncogenic ERG mediated transcription in prostate cells

The TMPRSS2/ERG gene rearrangement occurs in 50% of prostate tumors and results in expression of the transcription factor ERG, which is normally silent in prostate cells. ERG expression promotes prostate tumor formation and luminal epithelial cell fates when combined with PI3K/AKT pathway activation, however the mechanism of synergy is not known. In contrast to luminal fates, expression of ERG alone in immortalized normal prostate epithelial cells promotes cell migration and epithelial to mesenchymal transition (EMT). Migration requires ERG serine 96 phosphorylation via endogenous Ras/ERK signaling. We found that a phosphomimetic mutant, S96E ERG, drove tumor formation and clonogenic survival without activated AKT. S96 was only phosphorylated on nuclear ERG, and differential recruitment of ERK to a subset of ERG-bound chromatin associated with ERG-activated, but not ERG-repressed genes. S96E did not alter ERG genomic binding, but caused a loss of ERG-mediated repression, EZH2 binding and H3K27 methylation. In contrast, AKT activation altered the ERG cistrome and promoted expression of luminal cell fate genes. These data suggest that, depending on AKT status, ERG can promote either luminal or EMT transcription programs, but ERG can promote tumorigenesis independent of these cell fates and tumorigenesis requires only the transcriptional activation function.