Mycophenolate Mofetil (MYC) is a transplant drug used to prevent rejection in heart and kidneys transplant patients. Inosine monophosphate dehydrogenase (IMPDH), an enzyme involved in de novo synthesis of guanosine nucleotides, was considered as a primary target for MYC. Recently, we described that MYC was activates aryl hydrocarbon receptor and it antagonizes glucocorticoid receptor. Here we describe an androgen receptor (AR) as another off-target for MYC. We found that MYC increased basal and dihydrotestosterone (DHT)-inducible AR-dependent luciferase activity in AIZ-AR cells. In the same manner it induced or augmented mRNA level of KLK3 (prostate specific antigen; PSA) in 22Rv1 cells. Herein it displayed a hormetic effect on proliferation activity, since it significantly stimulated proliferation in lower concentrations but inhibited in higher (>1 µg/ml) concentrations in the presence of DHT. In contrast, MYC suppressed DHT-inducible KLK3 mRNA expression and cell proliferation in androgen-dependent LNCaP cells. MYC augmented DHT-inducible nuclear translocation of AR and increased the expression of MAPK8/9 (JNK46/54) resulting in the drop of their phosphorylation status. Moreover, MYC sensitized DHT-treated 22Rv1 cells to JNK-IN-8 mediated growth inhibition with the drop of IC50 from 1425 nM to 84 nM within 24 hrs. In conclusion, we suggest that, castrate-resistant prostate cancers progression might be retarded with the combination of MYC and chemical JNK inhibitors, involving AR-dependent mechanism.

• Publikační typ
• časopisecké články MeSH

The activation of the aryl hydrocarbon receptor (AhR) by xenobiotic compounds was demonstrated to result in the degradation of the androgen receptor (AR). Since prostate cancer is often dependent on AR, it has become a significant therapeutic target. As a result of the emerging concept of bacterial mimicry, we tested whether compounds with indole scaffolds capable of AhR activation have the potential to restrict AR activity in prostate cancer cells. Altogether, 22 indolic compounds were tested, and all of them activated AhR. However, only eight decreased DHT-induced AR luciferase activity. All indoles, which met the AhR-activating and AR-suppressing criteria, decreased the expression of DHT-inducible AR target genes, specifically KLK3 and FKBP5 mRNAs. The reduced AR binding to the KLK3 promoter was confirmed by a chromatin immunoprecipitation (ChIP) assay. In addition, some indoles significantly decreased AR protein and mRNA level. By using CRISPR/Cas9 AhR knockout technology, no relationship between AhR and AR, measured as target gene expression, was observed. In conclusion, some indoles that activate AhR possess AR-inhibiting activity, which seems to be related to the downregulation of AR expression rather than to AR degradation alone. Moreover, there does not seem to be a clear relationship that would connect AhR activation with AR activity suppression in 22Rv1 cells.

• MeSH
• androgenní receptory genetika   metabolismus   MeSH
• indoly farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty * genetika   metabolismus   MeSH
• receptory aromatických uhlovodíků * genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Současné možnosti diagnostiky karcinomu prostaty neumožňují odlišit agresivní (signifikantní tumory) od klinicky latentních (nesignifikantních forem). Práce se zaměřuje na stanovení hladiny klinicky využitelných tumorových markerů karcinomu prostaty: alfa-metyl CoA-racemáza (AMACR), caveolin-1, metalothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostatický specifický antigen (PSA), ZIP1 a ZnT-1 na úrovni prostatické tkáně (reprezentované buněčnými liniemi 22Rv1 a PNT1A) a v séru pacientů s histologicky ověřeným adenokarcinomem (82 karcinomových a 51 kontrolních sér). Na úrovni mRNA bylo zjištěno snížení exprese Cav-1, NF-?B, c-FOS a c-JUN a zvýšení exprese metalothioneinu, AMACR, PSA, Ki-67, MMP-9 a zinkových transportérů ZIP1 a ZnT-1. V séru je popsáno signifikantní zvýšení metalothioneinu nad hladinu 1 µM. Hladina caveolinu-1 se významně zvyšuje u high-grade tumorů, což jej staví do pozice možného markeru agresivní formy karcinomu prostaty.

Current methods for diagnosis of prostate carcinoma do not enable us to distinguish aggressive tumours (significant tumours) from clinically latent tumours (non-significant tumours). This study aims to determine levels of potential clinically important tumour markers such as alpha-methyl CoA-racemase (AMACR), caveolin-1, metallothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostate-specific antigen (PSA), ZIP1 and ZnT-1 in prostatic tissue represented by 22Rv1 (tumour) and PNT1A (healthy) cell lines and in blood serum of patients with histologically evaluated adenocarcinoma (82 tumour patients and 51 healthy volunteers). Based on mRNA expression, it was found that Cav-1, NF-?B, c-FOS and c-JUN were down-regulated and MT, AMACR, PSA, Ki-67, MMP-9 and zinc transporters ZIP1 and ZnT-1 were up-regulated. In serum, the level of MT was significantly enhanced above 1 µM. Caveolin-1 levels were significantly increased in high-grade tumours, which points to the possibility of using this protein as a marker for the aggressive form of prostate carcinoma.

• Klíčová slova
• nádorová onemocnění, imunodetekce, molekulárně-biologické techniky,
• MeSH
• 2D gelová elektroforéza využití   MeSH
• diagnostické techniky molekulární metody   MeSH
• elektrochemie metody   MeSH
• financování organizované MeSH
• kaveolin 1 diagnostické užití   MeSH
• lidé MeSH
• nádorové biomarkery krev   MeSH
• nádorové buněčné linie chemie   MeSH
• nádory prostaty genetika   krev   MeSH
• polymerázová řetězová reakce využití   MeSH
• prostatický specifický antigen izolace a purifikace   krev   MeSH
• senzitivita a specificita MeSH
• stupeň závažnosti nemoci MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH

MicroRNAs (miRNAs) are a large class of single-stranded RNA molecules involved in post-transcriptional gene silencing. miRNAs not only regulate various developmental and physiological processes but also are involved in cancer development. Additionally, they can be considered as biomarkers of some pathological processes. The aim of this study was to determine the expression levels of selected miRNA and zinc(II)-related genes (ZIP-1, BAX, MT2A and MT1A) in the non-tumor PNT1A prostate cell line in comparison with the prostate cancer cell lines 22Rv1, PC-3 and LNCaP after zinc(II) treatment. Using bioinformatic approaches we selected miRNAs with putative binding sites in the 3'UTR regions in Metallothionein 1A and 2A as miRNA 23a, 141, 224, 296-3p, 320, 375 and 376. We observed significantly higher expression of miRNA 23a in all tumor lines compared to non-tumor PNT1A (13.6-fold in 22Rv1, 7.3-fold in PC-3, 8.3-fold in LNCaP, p<0.01). We also observed that the 22Rv1 cell line has significantly higher expression of miRNA 224 in comparison to other cell lines. In addition, all tumor cell lines expressed significantly higher levels of miRNA 375 in comparison to non-tumor PNT1A (87.1‑fold in 22Rv1, nearly 2,000-fold in PC-3, 56.3‑fold in LNCaP, p<0.01). Nevertheless, miRNA 375 and 23a expression levels strongly suggest their potential to contribute to the diagnosis of prostate cancer and miRNA 224 eventually may be suitable for classification of primary tumors. The expression of miRNA 224 in 22Rv1 cell line was negatively correlated with increasing zinc(II) concentration only. Our experiments revealed significant negative correlation of miRNA 376 and MT2A in 22Rv1 and a negative correlation between miRNA 224 and MT1A in PC-3 cells which may denote possible direct regulation of MT genes by specific miRNAs in prostate cancer.

• MeSH
• dospělí MeSH
• genetická transkripce účinky léků   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• mikro RNA genetika   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty genetika   metabolismus   MeSH
• regulace genové exprese u nádorů * účinky léků   MeSH
• síran zinečnatý farmakologie   MeSH
• zinek metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Doxorubicin is an effective chemotherapeutic drug, however, its toxicity is a significant limitation in therapy. Encapsulation of doxorubicin inside liposomes or ferritin cages decreases cardiotoxicity while maintaining anticancer potency. We synthesized novel apoferritin- and liposome-encapsulated forms of doxorubicin ("Apodox" and "lip-8-dox") and compared its toxicity with doxorubicin and Myocet on prostate cell lines. Three different prostatic cell lines PNT1A, 22Rv1, and LNCaP were chosen. The toxicity of the modified doxorubicin forms was compared to conventional doxorubicin using the MTT assay, real-time cell impedance-based cell growth method (RTCA), and flow cytometry. The efficiency of doxorubicin entrapment was 56% in apoferritin cages and 42% in the liposome carrier. The accuracy of the RTCA system was verified by flow-cytometric analysis of cell viability. The doxorubicin half maximal inhibition concentrations (IC50) were determined as 170.5, 234.0, and 169.0 nM for PNT1A, 22Rv1, and LNCaP, respectively by RTCA. Lip8-dox is less toxic on the non-tumor cell line PNT1A compared to doxorubicin, while still maintaining the toxicity to tumorous cell lines similar to doxorubicin or epirubicin (IC50 = 2076.7 nM for PNT1A vs. 935.3 and 729.0 nM for 22Rv1 and LNCaP). Apodox IC50 was determined as follows: 603.1, 1344.2, and 931.2 nM for PNT1A, 22Rv1, and LNCaP.

• MeSH
• apoferritiny * MeSH
• doxorubicin aplikace a dávkování   chemie   toxicita   MeSH
• inhibiční koncentrace 50 MeSH
• koně MeSH
• lidé MeSH
• liposomy * MeSH
• nádorové buněčné linie MeSH
• nosiče léků MeSH
• příprava léků MeSH
• protinádorová antibiotika aplikace a dávkování   chemie   toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Androgen receptor (AR) signalling is triggered by androgens that have lipophilic nature. Since it was indicated that graphene oxide (GO) might facilitate passive diffusion of lipophilic compounds probably via Trojan horse-like mechanism, we tested the hypothesis if this suggestion would apply for androgens as well. Thus, we investigated if GO affects dihydrotestosterone (DHT)-triggered signalling of AR in two prostate cancer-derived cell lines, 22Rv1 and LNCaP. These cell lines differ in number of AR variants, i.e. there are two variants in 22Rv1 cells (full length and truncated) but only one in LNCaP cells (full length). Graphene oxide had no effect on basal luciferase activity but significantly decreased DHT-inducible AR-dependent luciferase activity in stably transfected cells. In 22Rv1 cells, it induced concentration-dependent decrease of DHT-inducible KLK3 mRNA and PSA protein after 24 h. While there was no effect on UBE2C mRNA (regulated by truncated variant), there was synergistic effect of DHT and GO on UBE2C protein level. Translocation of full-length AR (AR-FL) was potentiated by GO in the presence of DHT in 22Rv1 cells but it was suppressed in LNCaP cells. DHT-stimulated enrichment of AR-FL on KLK3 promoter was not significantly affected by GO in any tested cell line neither was KLK3 mRNA at 4 h of incubation. In conclusion, GO affects DHT-triggered signalling in both types of cells in similar manner, but ligand-triggered redistribution of AR-FL is affected differently. One of the reasons may be the presence of truncated variant of androgen receptor.

• MeSH
• androgenní receptory * genetika   MeSH
• androgeny MeSH
• dihydrotestosteron farmakologie   MeSH
• grafit MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

This study shows the effects of spices, and their phenolic and flavonoid compounds, on prostate cell lines (PNT1A, 22RV1 and PC3). The results of an MTT assay on extracts from eight spices revealed the strongest inhibitory effects were from black pepper and caraway seed extracts. The strongest inhibitory effect on prostatic cells was observed after the application of extracts of spices in concentration of 12.5 mg·mL-1. An LC/MS analysis identified that the most abundant phenolic and flavonoid compounds in black pepper are 3,4-dihydroxybenzaldehyde and naringenin chalcone, while the most abundant phenolic and flavonoid compounds in caraway seeds are neochlorogenic acid and apigenin. Using an MTT assay for the phenolic and flavonoid compounds from spices, we identified the IC50 value of ~1 mmol·L-1 PNT1A. The scratch test demonstrated that the most potent inhibitory effect on PNT1A, 22RV1 and PC3 cells is from the naringenin chalcone contained in black pepper. From the spectrum of compounds assessed, the naringenin chalcone contained in black pepper was identified as the most potent inhibitor of the growth of prostate cells.

• MeSH
• antikarcinogenní látky chemie   farmakologie   MeSH
• buněčné linie MeSH
• chromatografie kapalinová MeSH
• fenoly chemie   farmakologie   MeSH
• flavonoidy chemie   farmakologie   MeSH
• hmotnostní spektrometrie MeSH
• hojení ran účinky léků   MeSH
• koření analýza   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• prostata MeSH
• rostlinné extrakty chemie   farmakologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Sarcosine (N-methylglycine) was previously delineated as a substantial oncometabolite of prostate cancer (PCa) and its metabolism seems to be significantly involved in PCa development and behavior. METHODS: We focused on investigation whether the exposure of prostate cells (PNT1A, 22Rv1, and PC-3) to sarcosine-related amino acids (glycine, dimethylglycine, and sarcosine) affects their aggressiveness (cell mobility and division rates, using real-time cell based assay). The effect of supplementation on expression of glycine-N-methyltransferase (GNMT) mRNA was examined using qRT-PCR. Finally, post-treatment amino acids patterns were determined with consequent statistical processing using the Ward's method, factorial ANOVA and principal component analysis (P < 0.05). RESULTS: The highest migration induced sarcosine and glycine in metastatic PC-3 cells (a decrease in relative free area about 53% and 73%). The highest cell division was achieved after treatment of 22Rv1 and PC-3 cells with sarcosine (time required for division decreased by 65% or 45%, when compared to untreated cells). qRT-PCR revealed also significant effects on expression of GNMT. Finally, amino acid profiling shown specific amino acid patterns for each cell line. In both, treated and untreated PC-3 cells significantly higher levels of serine, glutamic acid, and aspartate, linked with prostate cancer progression were found. CONCLUSIONS: Sarcosine-related amino acids can exceptionally affect the behavior of benign and malignant prostate cells.

• MeSH
• aminokyseliny farmakologie   MeSH
• buněčné linie MeSH
• glycin-N-methyltransferasa genetika   metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty metabolismus   patologie   MeSH
• pohyb buněk účinky léků   MeSH
• prostata účinky léků   metabolismus   patologie   MeSH
• sarkosin metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Current diagnostic techniques are inefficient in distinguishing latent and low-risk forms of prostate cancer from high-risk forms. The present study is focused on determination of putative tumor markers of aggressive high-grade forms of prostate cancer. Potential markers were determined in blood sera of 133 patients (82 cases and 51 controls) and in cell lines (Gleason score 9-derived 22Rv1 and normal tissue derived PNT1A) on mRNA and protein levels. Alpha-methylacyl-CoA racemase (AMACR), metallothionein classes 1A and 2A (MT1A and MT2A) were determined and compared to prostate specific antigen (PSA) levels. On mRNA level, significantly increased expression of MT2A (2.4-fold), PSA (2.6-fold) and AMACR (8.4-fold) and insignificantly (1.9-fold) elevated MT1A in 22Rv1 compared to non-tumor PNT1A were determined. On protein level, significant enhancement of free PSA and total PSA in tumor cell line was evident. AMACR protein was 1.5-fold elevated in tumor line (below the level of significance). Contrary to mRNA, significantly (p = 0.01) reduced level of MT protein in tumor lines was determined. In the case of serum level, significantly enhanced MT level (4.5-fold) in patients' sera was found. No significant changes were observed in the case of AMACR. These findings indicate possible alternative role of MT to PSA prostate cancer marker. In addition, level of AMACR is distinctly higher in the Gleason score 9 in serum of patients and MT shows a descending trend in relation to Gleason score.

• MeSH
• adenokarcinom genetika   metabolismus   patologie   MeSH
• dospělí MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• metalothionein genetika   metabolismus   MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nádory prostaty genetika   metabolismus   patologie   MeSH
• prognóza MeSH
• prostatický specifický antigen genetika   metabolismus   MeSH
• racemasy a epimerasy genetika   metabolismus   MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• stupeň nádoru MeSH
• western blotting MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Prostate cancer (PCa) is the most common cancer and the second leading cause of cancer-related deaths of men in Western countries. Androgen deprivation therapy is initially successful, however eventually fails, and tumors progress to the more aggressive castration-resistant PCa (CRPC). Yet, androgen receptor (AR) usually remains as a major regulator of tumor cell proliferation in CRPC. Interleukin-23 (IL-23) was recently shown to promote the development of CRPC by driving AR transcription. Here we used the androgen-sensitive LNCaP, castration-resistant C4-2, and 22Rv1 cells. Interestingly, cellular senescence is induced in these human cell lines by treatment with the AR antagonists enzalutamide (ENZ) or darolutamide (ODM), which might be one underlying mechanism for inhibition of PCa cell proliferation. Treatment with IL-23 alone did not change cellular senescence levels in these cell lines, whereas IL-23 inhibited significantly cellular senescence levels induced by ENZ or ODM in both CRPC cell lines C4-2 and 22Rv1 but not in LNCaP cells. This indicates a response of IL-23 specific in CRPC cells. Generating LNCaP and C4-2 three-dimensional (3D) spheroids and treatment with AR antagonists resulted in the reduced spheroid volume and thus growth inhibition. However, the combination of AR antagonists with IL-23 did not affect the antagonist-mediated reduction of spheroid volumes. This observation was confirmed with proliferation assays using adherent monolayer cell cultures. Taken together, the data indicate that IL-23 treatment reduces the AR antagonists-induced level of cellular senescence of CRPC cells, which could be one possible mechanism for promoting castration resistance.

• MeSH
• antagonisté androgenních receptorů farmakologie   terapeutické užití   MeSH
• benzamidy farmakologie   terapeutické užití   MeSH
• fenylthiohydantoin farmakologie   terapeutické užití   MeSH
• interleukin-23 metabolismus   MeSH
• lidé MeSH
• nitrily farmakologie   terapeutické užití   MeSH
• protokoly protinádorové kombinované chemoterapie farmakologie   terapeutické užití   MeSH
• pyrazoly farmakologie   terapeutické užití   MeSH
• stárnutí buněk MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH