Cisplatin is widely used as an antitumor drug. To reduce its toxic side effects in patients, cisplatin has been bound with procaine in a cisplatin-procaine complex (DPR). The lethal and teratogenic effects of cisplatin alone and of complexed cisplatin were determined in the chick embryo in ovo in order to compare their influence on rapidly proliferating embryonic tissues. The embryotoxic (lethal + teratogenic) effect was examined after a single intra-amniotic injection of one of six different doses, ranging from 0.03 to 30.0 microg, on embryonic days (ED) 3, 4 or 5. The minimal embryotoxic dose was lower for cisplatin alone (0.03-0.3 microg) than for cisplatin in the DPR complex (0.3-3.0 microg), suggesting that cisplatin alone is more embryotoxic than complexed cisplatin. Both substances caused malformations in the surviving embryos evaluated on ED 9. These malformations included microphthalmia, microcephaly, hypoplasia of the upper and lower jaw, cleft beak, and haemocephaly. Moreover, heart septum defects and limb reduction deformities were found after exposure to the DPR complex. The embryotoxicity of complexed cisplatin exhibited a stage-response effect. It was highest on day 3 and gradually decreased until ED 5. Such an apparent stage-response effect was not observed for cisplatin alone. The embryotoxicity of procaine hydrochloride - a component of the complex - was also tested. Procaine hydrochloride alone did not produce any embryotoxic effect, not even after a single injection of the maximal tested dose (100.0 microg per embryo). We also examined the protective effect of procaine hydrochloride, whose separate administration at ED 4 was followed by the injection of 0.3 microg cisplatin. We did not observe any protective effect of procaine hydrochloride if injected separately.

• MeSH
• abnormality vyvolané léky etiologie   MeSH
• anestetika lokální aplikace a dávkování   farmakologie   MeSH
• antitumorózní látky aplikace a dávkování   toxicita   MeSH
• cisplatina aplikace a dávkování   analogy a deriváty   toxicita   MeSH
• kuřecí embryo abnormality   účinky léků   MeSH
• organoplatinové sloučeniny aplikace a dávkování   toxicita   MeSH
• prokain aplikace a dávkování   analogy a deriváty   farmakologie   toxicita   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo abnormality   účinky léků   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• anestetika lokální MeSH
• antitumorózní látky MeSH
• cisplatin-procaine complex MeSH Prohlížeč
• cisplatina MeSH
• organoplatinové sloučeniny MeSH
• prokain MeSH

AIMS: Cisplatin is a widely used chemotherapeutic. However, it is associated with numerous adverse effects. The aim of our study was examination of cisplatin interaction with Na(+)/K(+)-ATPase (NKA, the sodium pump). This enzyme is of crucial importance for all animal cells and particularly for the kidney, which is frequently damaged during chemotherapy. METHODS: The entire NKA was isolated from porcine kidney. Its large cytoplasmic segment connecting transmembrane helices 4 and 5 (C45), was heterologously expressed in E.coli (wild-type or C367S mutant). The ATPase activity was evaluated according to the inorganic phosphate production and the interaction of isolated C45 with cisplatin was studied using chronopotentiometry and mass spectrometry. RESULTS: Our experiments revealed that cisplatin can inhibit NKA. The finding that other platinum-based drugs with a low nephrotoxicity, carboplatin and oxaliplatin, did not inhibit NKA, suggested that NKA/cisplatin interaction is an important factor in cisplatin adverse effects. The inhibitory effect of cisplatin could be prevented by preincubation of the enzyme with reduced glutathione or DTT. Using chronopotentiometry and mass spectrometry, we found that cisplatin is bound to C45. However, our mutagenesis experiment did not confirm that the suggested Cys367 could be the binding site for cisplatin. CONCLUSION: Unintended interactions of drugs present serious limitations to treatment success. Although a large number of membrane pumps have been identified as potential targets of cisplatin, vis-a-vis nephrotoxicity, NKA inhibition seems to be of crucial importance. Experiments with isolated large cytoplasmic segment C45 revealed that it is the main target of cisplatin on NKA and that the reaction with cysteine residues plays an important role in cisplatin/NKA interactions. However, further experiments must be performed to identify the interacting amino acid residues more precisely.

• MeSH
• antitumorózní látky škodlivé účinky   chemie   farmakologie   MeSH
• cisplatina škodlivé účinky   chemie   farmakologie   MeSH
• ledviny enzymologie   MeSH
• prasata MeSH
• sodíko-draslíková ATPasa antagonisté a inhibitory   chemie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antitumorózní látky MeSH
• cisplatina MeSH
• sodíko-draslíková ATPasa MeSH

Cisplatin is the most widely used chemotherapeutics for cancer treatment, however, its administration is connected to inevitable adverse effects. Previous studies suggested that cisplatin is able to inhibit Na(+)/K(+)-ATPase (NKA), the enzyme responsible for maintaining electrochemical potential and sodium gradient across the plasma membrane. Here we report a crystallographic analysis of cisplatin bound to NKA in the ouabain bound E2P form. Despite a moderate resolution (7.4 Å and 7.9 Å), the anomalous scattering from platinum and a model representation from a recently published structure enabled localization of seven cisplatin binding sites by anomalous difference Fourier maps. Comparison with NKA structures in the E1P conformation suggested two possible inhibitory mechanisms for cisplatin. Binding to Met151 can block the N-terminal pathway for transported cations, while binding to Met171 can hinder the interaction of cytoplasmic domains during the catalytic cycle.

• Klíčová slova
• Adverse effects, Binding sites, Cisplatin, Na(+)/K(+)-ATPase, Sodium pump, X-ray crystallography,
• MeSH
• cisplatina škodlivé účinky   chemie   metabolismus   MeSH
• konformace proteinů MeSH
• krystalografie rentgenová MeSH
• methionin chemie   metabolismus   MeSH
• molekulární modely MeSH
• ouabain chemie   MeSH
• sodíko-draslíková ATPasa antagonisté a inhibitory   chemie   metabolismus   MeSH
• terciární struktura proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cisplatina MeSH
• methionin MeSH
• ouabain MeSH
• sodíko-draslíková ATPasa MeSH

The efficiency of cisplatin (CDDP) is significantly hindered by the development of resistance during the treatment course. To gain a detailed understanding of the molecular mechanisms underlying the development of cisplatin resistance, we comparatively analyzed established a CDDP-resistant neuroblastoma cell line (UKF-NB-4CDDP) and its susceptible parental cells (UKF-NB-4). We verified increased chemoresistance of UKF-NB-4CDDP cells by analyzing the viability, induction of apoptosis and clonal efficiency. To shed more light on this phenomenon, we employed custom cDNA microarray (containing 2234 probes) to perform parallel transcriptomic profiling of RNA and identified that 139 genes were significantly up-regulated due to CDDP chemoresistance. The analyses of molecular pathways indicated that the top up-regulation scoring functions were response to stress, abiotic stimulus, regulation of metabolic process, apoptotic processes, regulation of cell proliferation, DNA repair or regulation of catalytic activity, which was also evidenced by analysis of molecular functions revealing up-regulation of genes encoding several proteins with a wide-spectrum of enzymatic activities. Functional analysis using lysosomotropic agents chloroquine and bafilomycin A1 validated their potential to re-sensitize UKF-NB-4CDDP cells to CDDP. Taken together, the identification of alterations in specific genes and pathways that contribute to CDDP chemoresistance may potentially lead to a renewed interest in the development of novel rational therapeutics and prognostic biomarkers for the management of CDDP-resistant neuroblastoma.

• Klíčová slova
• chemoresistance, cisplatin, lysosomes, microarray, neuroblastoma, transport,
• MeSH
• buněčné klony MeSH
• chemorezistence účinky léků   genetika   MeSH
• chlorochin farmakologie   MeSH
• cisplatina farmakologie   terapeutické užití   MeSH
• genová ontologie MeSH
• genové regulační sítě účinky léků   MeSH
• lidé MeSH
• lyzozomy účinky léků   metabolismus   MeSH
• makrolidy farmakologie   MeSH
• nádorové buněčné linie MeSH
• neuroblastom farmakoterapie   genetika   patologie   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• transkriptom účinky léků   genetika   MeSH
• tvar buňky účinky léků   MeSH
• upregulace účinky léků   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bafilomycin A1 MeSH Prohlížeč
• chlorochin MeSH
• cisplatina MeSH
• makrolidy MeSH

Patients with testicular tumors receiving the Einhorn combination chemotherapy with cisplatinum administration for five consecutive days were evaluated for signs of glomerular filtration rate impairment by means of serum beta 2-microglobulin measurements before and on days 2 and 5 after cessation of cisplatin administration and then before repeated cycles of the same chemotherapeutic regimen. Though no significant changes in the level of serum beta 2-microglobulin could be found, an evident increase in the values was seen indicating that in more aggressive cisplatin administration schedules not only tubular impairment might contribute to possible nephrotoxic effects.

• MeSH
• beta-2-mikroglobulin analýza   MeSH
• cisplatina škodlivé účinky   MeSH
• ledviny účinky léků   MeSH
• lidé MeSH
• testikulární nádory farmakoterapie   MeSH
• vyšetření funkce ledvin MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta-2-mikroglobulin MeSH
• cisplatina MeSH

Neurotoxic effects of cisplatin and the cisplatin-procaine complex cis-diaminechloro-[2-(diethylamino)ethyl 4-aminobenzoate, N4]-chlorideplatinum(II) monohydrochloride monohydrate (DPR) were compared in organotypic cultures of chick embryonic dorsal root ganglia maintained in a semi-solid (soft agar) culture medium. The changes of two characteristics of the neurite outgrowth, the mean radial length of neuritic processes growing out from the ganglia and the area of neurite outgrowth around the ganglion, were used as parameters to evaluate the toxic effect of both compounds. The drugs were administered to the cultures at concentrations ranging from 13 to 120 microM. The half-maximum inhibition concentration (IC50) was determined from the concentration-response curves for both the mean radial length of neurites and the area of neurite outgrowth. An analysis of these parameters revealed that DPR was significantly less neurotoxic than cisplatin. In fact, considering the mean radial length of neurite processes, the IC50s of cisplatin were 56, 65 and 66 microM after 24, 48 and 72 h of exposure, respectively. By contrast, for DPR the IC50s were 116 microM after 24 h, and greater than 120 microM after 48 and 72 h of exposure. When we considered the area index (i.e. the area of neurite outgrowth normalized for the area of the ganglia), the IC50s for cisplatin were 41, 52 and 55 microM after 24, 48 and 72 h of exposure, respectively, whereas for DPR the IC50s were 59 microM after 24 h, and greater than 120 microM after 48 and 72 h of exposure. Our results support previous findings of lower toxicity of DPR to non-neoplastic tissues, as compared to cisplatin.

• MeSH
• antitumorózní látky aplikace a dávkování   škodlivé účinky   MeSH
• cisplatina aplikace a dávkování   škodlivé účinky   analogy a deriváty   MeSH
• inhibiční koncentrace 50 MeSH
• kuřecí embryo MeSH
• organoplatinové sloučeniny aplikace a dávkování   škodlivé účinky   MeSH
• prokain aplikace a dávkování   škodlivé účinky   analogy a deriváty   MeSH
• spinální ganglia účinky léků   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antitumorózní látky MeSH
• cisplatin-procaine complex MeSH Prohlížeč
• cisplatina MeSH
• organoplatinové sloučeniny MeSH
• prokain MeSH

Chemoresistance is a major problem in successful cancer therapy. Lysine-specific demethylase 5B (KDM5B), is a member of the KDM5 family of histone demethylases, whose dysregulation has been observed in numerous types of cancer and plays a role in drug tolerance. The present study examined KDM5B expression in high risk neuroblastoma cell lines. Its level was markedly reduced in cisplatin-resistant cells, UKF-NB-4CDDP, compared with parental sensitive cells UKF-NB-4. Moreover, KDM5B-silencing did not affect either viability nor the response to CDDP in resistant cells, and led to increase of proliferation and migration in CDDP resistant cells but not in sensitive ones. Compliant with these results, short interfering KDM5B transfection resulted in increased S phase in resistant cells. Overall, these findings suggested that KDM5B may be involved in the survival mechanisms of neuroblastoma cells, which makes KDM5B a promising factor for the prediction of sensitivity to CDDP that should therefore be considered for future research.

• Klíčová slova
• chemoresistance, cisplatin, histone methylation, lysine-specific demethylase 5B, neuroblastoma,
• Publikační typ
• časopisecké články MeSH

Cisplatin is a widely used chemotherapeutic agent that is clinically approved to fight both carcinomas and sarcomas. It has relatively high efficiency in treating ovarian cancers and metastatic testicular cancers. It is generally accepted that the major mechanism of cisplatin anti-cancer action is DNA damage. However, cisplatin is also effective in metastatic cancers and should, therefore, affect slow-cycling cancer stem cells in some way. In this review, we focused on the alternative effects of cisplatin that can support a good therapeutic response. First, attention was paid to the effects of cisplatin at the cellular level such as changes in intracellular pH and cellular mechanical properties. Alternative cellular targets of cisplatin, and the effects of cisplatin on cancer cell metabolism and ER stress were also discussed. Furthermore, the impacts of cisplatin on the tumor microenvironment and in the whole organism context were reviewed. In this review, we try to reveal possible causes of the unexpected effectiveness of this anti-cancer drug.

• MeSH
• antitumorózní látky farmakologie   terapeutické užití   MeSH
• biomechanika účinky léků   MeSH
• cisplatina farmakologie   terapeutické užití   MeSH
• koncentrace vodíkových iontů účinky léků   MeSH
• lidé MeSH
• metabolické sítě a dráhy účinky léků   MeSH
• nádorové mikroprostředí účinky léků   MeSH
• nádory farmakoterapie   metabolismus   patologie   MeSH
• stres endoplazmatického retikula účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• antitumorózní látky MeSH
• cisplatina MeSH

Metallothionein-3 has poorly characterized functions in neuroblastoma. Cisplatin-based chemotherapy is a major regimen to treat neuroblastoma, but its clinical efficacy is limited by chemoresistance. We investigated the impact of human metallothionein-3 (hMT3) up-regulation in neuroblastoma cells and the mechanisms underlying the cisplatin-resistance. We confirmed the cisplatin-metallothionein complex formation using mass spectrometry. Overexpression of hMT3 decreased the sensitivity of neuroblastoma UKF-NB-4 cells to cisplatin. We report, for the first time, cisplatin-sensitive human UKF-NB-4 cells remodelled into cisplatin-resistant cells via high and constitutive hMT3 expression in an in vivo model using chick chorioallantoic membrane assay. Comparative proteomic analysis demonstrated that several biological pathways related to apoptosis, transport, proteasome, and cellular stress were involved in cisplatin-resistance in hMT3 overexpressing UKF-NB-4 cells. Overall, our data confirmed that up-regulation of hMT3 positively correlated with increased cisplatin-chemoresistance in neuroblastoma, and a high level of hMT3 could be one of the causes of frequent tumour relapses.

• MeSH
• chemorezistence účinky léků   genetika   MeSH
• cisplatina farmakologie   MeSH
• kuřecí embryo MeSH
• lidé MeSH
• metalothionein 3 biosyntéza   genetika   MeSH
• nádorové buněčné linie MeSH
• nádorové proteiny biosyntéza   genetika   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cisplatina MeSH
• metalothionein 3 MeSH
• nádorové proteiny MeSH

Ototoxic effects of cisplatin (1.5 mg/kg), cisplatin with mannitol (1.5 mg/kg cisplatin with 15 mg/kg mannitol) and carboplatin (6 mg/kg) were compared in guinea pigs treated 5 days a week for 5 consecutive weeks. Auditory thresholds were measured in awake animals by means of the compound action potential recorded from electrodes implanted near the round window. The endocochlear potential (EP) was measured in the first turn through the round window, and hair cell counts were estimated by the surface specimen technique. The most pronounced ototoxicity (threshold shifts and hair cell loss) was observed with pure cisplatin. Cisplatin with mannitol appeared to be less toxic than pure cisplatin, and carboplatin the least toxic. Even carboplatin produced significant threshold shifts, particularly at high frequencies, but they were not accompanied by hair cell loss. Values of EP were not different from those in normal control animals.

• MeSH
• akustická stimulace MeSH
• cisplatina aplikace a dávkování   škodlivé účinky   farmakologie   MeSH
• fixní kombinace léků MeSH
• implantované elektrody MeSH
• injekce intraperitoneální MeSH
• karboplatina aplikace a dávkování   škodlivé účinky   farmakologie   MeSH
• kochlea účinky léků   MeSH
• mannitol MeSH
• morčata MeSH
• sluch * MeSH
• sluchový práh MeSH
• vláskové buňky účinky léků   MeSH
• vnitřní ucho účinky léků   MeSH
• zvířata MeSH
• Check Tag
• morčata MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cisplatina MeSH
• fixní kombinace léků MeSH
• karboplatina MeSH
• mannitol MeSH