Malignant cell proliferation and accumulation depends on the balance between the rates of cell production and cell death. Recent evidence indicates that apoptosis is important in the development of cancer. Apoptosis is strictly controlled by various regulators, which can take part in the apoptotic process, proliferation and differentiation alike. Apoptosis was induced in myeloid cell line ML-2 by camptothecin, an inhibitor of topoisomerase I. After 18 hours of induction by camptothecin 50% of cells were apoptotic. The apoptotic effect of CAM was reversible in the cells studied. The induction of apoptosis influenced the expression of apoptosis and cell cycle regulators as detected by cDNA arrays, RT-PCR or Western blotting. According to cDNA arrays e.g. bax, bfl1, bak, pRb2, c-jun, jun-B were upregulated, and cdk4, cyclin B1, wee1, CRAF1, DP1 were downregulated. A number of other regulators like p21 and cdc25A, as well as some other genes linked with apoptosis, as p53 and the bcl-2 family, were up- or down-regulated as determined by real-time PCR. Changes in gene expression were found not only in the group of regulators of apoptosis and the cell cycle, but also among regulators of differentiation.

• MeSH
• apoptóza účinky léků   genetika   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné dělení účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• down regulace MeSH
• fytogenní protinádorové látky farmakologie   MeSH
• kamptothecin farmakologie   MeSH
• lidé MeSH
• myeloidní leukemie genetika   patologie   MeSH
• nádorové buňky kultivované MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• stanovení celkové genové exprese * MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fytogenní protinádorové látky MeSH
• kamptothecin MeSH

Camptothecin (CPT), an alkaloid, was first discovered from plants and has potent anti-tumor activity. Since then, CPT analogs (namely Irinotecan and Topotecan) have been approved by the FDA for cancer treatments. Curcumin, on the other hand, is a widely used photosensitizer in photodynamic therapy (PDT) treatment. In our previous work, we have reported a straightforward strategy to construct a drug self-delivery system in which two-molecular species Irinotecan and Curcumin can self-assembly into a complex of ion pairs, namely ICN, through intermolecular non-covalent interactions. We found that ICN has slightly better chemotherapy efficacy than its individual components with much fewer side effects. In this paper, we aim to combine the chemotherapy and the PDT of ICN to further improve its anti-tumor performance. The efficient cellular uptake of ICNs was observed by confocal microscopy. Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay was used to detect the generation of singlet oxygen species. We found that the cell viability was 9% with both chemotherapy and PDT, and 31% with chemotherapy alone for the case with an ICN concentration of 10 μM, which demonstrated that the anti-tumor efficacy against the HT-29 cancer cell line was enhanced substantially with the combination therapy strategy. The study with an in vivo mouse model has further verified that the chemo-PDT dual therapy can inhibit tumor growth by 84% and 18.8% comparing with the control group and the chemotherapy group, respectively. Our results demonstrated that the new strategy using self-assembly and carrier-free nanoparticles with their chemo-PDT dual therapy may provide new opportunities to develop future combinatorial therapy methods in treating cancer.

• Klíčová slova
• Anti-tumor efficacy, Combination therapy, Drug delivery system, Photodynamic therapy,
• MeSH
• apoptóza účinky léků   účinky záření   MeSH
• buňky HT-29 MeSH
• diarylheptanoidy chemie   MeSH
• fotochemoterapie metody   MeSH
• intracelulární prostor účinky léků   metabolismus   účinky záření   MeSH
• kamptothecin chemie   farmakologie   MeSH
• kombinovaná terapie MeSH
• lidé MeSH
• protinádorové látky chemie   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• diarylheptanoidy MeSH
• kamptothecin MeSH
• protinádorové látky MeSH

Various DNA-targeting agents may initiate p53-dependent as well as p53-independent response and subsequent apoptosis via alternative cellular systems which include for instance p73, caspase-2 or Bcl-2 family proteins. The scope of involvement of individual molecules in this process and the mechanisms governing their potential interplay are still not entirely understood, in particular in highly aggressive cancers such as in malignant melanoma. In this work we investigated the role and involvement of both p53-dependent and -independent mechanisms in selected melanoma cell lines with differing status of p53 using a model DNA topoisomerase I inhibitor camptothecin (CPT). Here we report that CPT induced in Bowes melanoma cells apoptosis which is essentially p53 and mitochondria-dependent but with some involvement of caspase-2 and p73. Conversely, in mutant p53 melanoma cells overall levels of CPT-induced apoptosis are significantly lower, with p73 and caspase-2 signaling playing important roles. In addition, in these cells the expression of micro RNAs family 34 (miR-34) were low compared to wild-type p53 cells. The ectopic expression of wild type p53 than restored apoptotic response of cells to CPT despite the fact that the expression of miR-34 and miR-155 were not influenced. These results suggest that CPT induces multivariate cellular stress responses including activation of DNA-damage response-p53 pathway as well as p53-independent signaling and their mutual crosstalk play the decisive role in the efficient triggering of apoptosis in melanoma cells.

• MeSH
• apoptóza účinky léků   MeSH
• cysteinové endopeptidasy genetika   metabolismus   MeSH
• DNA vazebné proteiny genetika   metabolismus   MeSH
• fytogenní protinádorové látky farmakologie   MeSH
• inhibitory topoisomerasy I farmakologie   MeSH
• jaderné proteiny genetika   metabolismus   MeSH
• kamptothecin farmakologie   MeSH
• kaspasa 2 genetika   metabolismus   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• melanom genetika   metabolismus   patologie   MeSH
• mikro RNA genetika   metabolismus   MeSH
• mitochondrie účinky léků   genetika   metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové supresorové proteiny genetika   metabolismus   MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• poškození DNA účinky léků   MeSH
• protein p73 MeSH
• protoonkogenní proteiny c-bcl-2 genetika   metabolismus   MeSH
• signální transdukce * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CASP2 protein, human MeSH Prohlížeč
• cysteinové endopeptidasy MeSH
• DNA vazebné proteiny MeSH
• fytogenní protinádorové látky MeSH
• inhibitory topoisomerasy I MeSH
• jaderné proteiny MeSH
• kamptothecin MeSH
• kaspasa 2 MeSH
• mikro RNA MeSH
• nádorové supresorové proteiny MeSH
• nádorový supresorový protein p53 MeSH
• protein p73 MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• TP73 protein, human MeSH Prohlížeč

The mixed dissociation constants of four anticancer drugs--camptothecine, 7-ethyl-10-hydroxycamptothecine, 10-hydroxycamptothecine and 7-ethylcamptothecine, including diprotic and triprotic molecules at various ionic strengths I of range 0.01 and 0.4, and at temperatures of 25 and 37 degrees C--were determined with the use of two different multiwavelength and multivariate treatments of spectral data, SPECFIT32 and SQUAD(84) nonlinear regression analyses and INDICES factor analysis. A proposed strategy for dissociation constants determination is presented on the acid-base equilibria of camptothecine. Indices of precise modifications of the factor analysis in the program INDICES predict the correct number of components, and even the presence of minor ones, when the data quality is high and the instrumental error is known. The thermodynamic dissociation constant pK(a)(T) was estimated by nonlinear regression of {pK(a), I} data at 25 and 37 degrees C: for camptothecine pK(a,1)(T)=2.90(7) and 3.02(8), pK(a,2)(T)=10.18(30) and 10.23(8); for 7-ethyl-10-hydroxycamptothecine, pK(a,1)(T)=3.11(2) and 2.46(6), pK(a,2)(T)=8.91(4) and 8.74(3), pK(a,3)(T)=9.70(3) and 9.47(8); for 10-hydroxycamptothecine pK(a,1)(T)=2.93(4) and 2.84(5), pK(a,2)(T)=8.93(2) and 8.92(2), pK(a,3)(T)=9.45(10) and 9.98(4); and for 7-ethylcamptothecine pK(a,1)(T)=3.10(4) and 3.30(16), pK(a,2)(T)=9.94(9) and 10.98(18). Goodness-of-fit tests for various regression diagnostics enabled the reliability of the parameter estimates found to be proven. Pallas and Marvin predict pK(a) being based on the structural formulae of drug compounds in agreement with the experimental value.

• MeSH
• fytogenní protinádorové látky chemie   MeSH
• irinotekan MeSH
• kamptothecin analogy a deriváty   chemie   MeSH
• koncentrace vodíkových iontů MeSH
• metoda nejmenších čtverců MeSH
• spektrofotometrie ultrafialová MeSH
• termodynamika MeSH
• titrace MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 10-hydroxycamptothecin MeSH Prohlížeč
• 7-ethylcamptothecin MeSH Prohlížeč
• fytogenní protinádorové látky MeSH
• irinotekan MeSH
• kamptothecin MeSH

Various design and fabrication strategies of carrier-based drug delivery systems have been quickly established and applied for cancer therapy in recent years. These systems contribute greatly to current cancer treatments but further development needs to be made to eliminate obstacles such as low drug loading capacity and severe side effects. To achieve better drug delivery, we propose an innovative strategy for the construction of easy manufactured drug self-delivery systems based on molecular structures, which can be used for the co-delivery of curcuminoids and all the nitrogen-containing derivatives of camptothecin for better targeted cancer therapy with minimized side effects. The formation mechanism investigation demonstrates that the rigid planar structures of camptothecin derivatives and curcuminoids with relevant leaving hydrogens make it possible for them to be assembled into nanoparticles under suitable conditions. These nanoparticles show stabilized particle sizes (100 nm) under various conditions and tunable surface charges which increase from around -10 mV in a normal physiological condition (pH 7.4) to +40 mV under acidic tumor environments. In addition, in vivo mice experiments have demonstrated that, compared to irinotecan (a derivative of camptothecin) itself, the co-delivered irinotecan curcumin nanoparticles exhibited significantly enhanced lung and gallbladder targeting, improved macrophage-clearance escape and ameliorated colorectal cancer treatment with an eradication of life-threatening diarrhea, bringing hope for better targeted chemotherapy and clinical translation. Lastly, the strategy of structure based design of drug self-delivery systems may inspire more research and discoveries of similar self-delivered nano systems for wider pharmaceutical applications.

• Klíčová slova
• Charge conversion, Curcumin, Diarrhea, Irinotecan, Self-delivery, Targeting therapy,
• MeSH
• kamptothecin MeSH
• léčivé přípravky * MeSH
• lékové transportní systémy * MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory * farmakoterapie   MeSH
• nanočástice * MeSH
• protinádorové látky * terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kamptothecin MeSH
• léčivé přípravky * MeSH
• protinádorové látky * MeSH

Yeast cell viability was evaluated microscopically following exposure to heat shock for 30 min at 53 degrees C. The cells were previously grown in the presence of potential stressors (anticancer drugs; e.g., 5-fluorouracil, methotrexate, cisplatin, bleomycin, mitomycin-C and camptothecin-11). The induction of thermotolerance was documented by significantly increased viability after heat shock. This effect, which was reversed by cycloheximide, was comparable to that observed following exposure to a mild heat stress. These data demonstrate that pretreatment with sub-toxic concentrations of some of the clinically used antineoplastic agents conferres thermotolerance to yeast, possibly through the synthesis of protein components.

• MeSH
• bleomycin farmakologie   MeSH
• cisplatina farmakologie   MeSH
• fluoruracil farmakologie   MeSH
• irinotekan MeSH
• kamptothecin analogy a deriváty   farmakologie   MeSH
• methotrexát farmakologie   MeSH
• mitomycin farmakologie   MeSH
• protinádorové látky farmakologie   MeSH
• Saccharomyces účinky léků   fyziologie   MeSH
• vysoká teplota * MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bleomycin MeSH
• cisplatina MeSH
• fluoruracil MeSH
• irinotekan MeSH
• kamptothecin MeSH
• methotrexát MeSH
• mitomycin MeSH
• protinádorové látky MeSH

Caspases are key enzymatic components of the intracellular apoptotic machinery, and their role in mammalian systems is often studied using fluoromethylketone (FMK) inhibitors. Despite many advantages of such approach, efficiency of the inhibitor and membrane permeability speed are often questioned. This work therefore focuses on an exact evaluation of caspase-3 FMK inhibition dynamics in camptothecin-induced mesenchymal micromasses. Two parameters of caspase-3 FMK inhibitor were investigated: first, the stability of the inhibitory potential in the time course of cultivation and, simultaneously, the dynamics of caspase-3 FMK inhibition after camptothecin-induced apoptosis peak. A photon-counting chemiluminescence approach was applied for quantification of active caspase-3. The sensitivity of the photon-counting method allowed for evaluation of active caspase-3 concentration in femtogram amounts per cell. The inhibitor penetrated the cells within the first minute after its application, and the peak of caspase-3 started to decline to the blank level after 30 min. The inhibitory effect of the FMK inhibitor was unchanged during the entire 48 h of cultivation.

• MeSH
• apoptóza účinky léků   MeSH
• inbrední kmeny myší MeSH
• inhibitory kaspas farmakologie   MeSH
• kamptothecin farmakologie   MeSH
• kaspasa 3 metabolismus   fyziologie   MeSH
• kultivované buňky MeSH
• luminiscenční měření metody   MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• inhibitory kaspas MeSH
• kamptothecin MeSH
• kaspasa 3 MeSH

The colorectal cancer ranks high among the malignant tumours in incidence and mortality and irinotecan is standardly used in palliative treatment of metastatic disease in every therapeutic line. Unfortunately, the treatment with irinotecan is often associated with severe toxicities, especially neutropenia and diarrhea. The majority of the toxic manifestation is caused by the insufficient deactivation (glucuronidation) of irinotecan active metabolite SN-38 by UGT1A enzyme. The elevated SN-38 plasma concentration is responsible for the hematological and gastrointestinal toxicity that can become life-threatening. The patients carrying the mutation of the gene encoding UGT1A enzyme lack the ability of bilirubin glucuronidation, and suffer from the inherited un-conjugated hyperbilirubinemia (Gilbert syndrome, Crigler-Najjar type 1 and 2 syndrome). The mutations in other enzyme systems also play role in the etiopathogenesis of the irinotecan toxicity: CYP3A (cytochrome P-450), ABC family of transmembrane transporters (adenosine-triphosphate binding cassette). The goal of the contemporary research is to determine the predictive factors that will enable the individual adjustment of the individual drug dosage while minimising the adverse effects and maintaining the treatment benefit.

• MeSH
• farmakogenetika MeSH
• fytogenní protinádorové látky škodlivé účinky   farmakokinetika   terapeutické užití   MeSH
• irinotekan MeSH
• kamptothecin škodlivé účinky   analogy a deriváty   farmakokinetika   terapeutické užití   MeSH
• kolorektální nádory genetika   patologie   MeSH
• lidé MeSH
• neutropenie chemicky indukované   MeSH
• průjem chemicky indukované   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• fytogenní protinádorové látky MeSH
• irinotekan MeSH
• kamptothecin MeSH

The dynamics of cell morphology, in particular membrane blebbing, was studied after induction of apoptosis by etoposide or camptothecin in four human stabilized cell lines (Hep2, Bowes, HT-29 and HL-60). Time lapse videomicroscopy and F-actin staining revealed various dynamic parameters of this process including its duration, maximal extent, stages and final endpoints in individual cells. Although generally occurring in predictable patterns, our results indicate a relatively significant variability both in appearance and temporal organization of blebbing not only between different cell lines but also within them.

• MeSH
• apoptóza účinky léků   MeSH
• buněčná membrána diagnostické zobrazování   účinky léků   MeSH
• cytoskelet účinky léků   MeSH
• etoposid farmakologie   MeSH
• kamptothecin farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie diagnostické zobrazování   účinky léků   MeSH
• protinádorové látky farmakologie   MeSH
• ultrasonografie MeSH
• videomikroskopie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• etoposid MeSH
• kamptothecin MeSH
• protinádorové látky MeSH

DNA damage in embryos shapes the development of an organism. Understanding life stage-specific differences between fish species is essential for ecological risk assessment measures. We explored DNA damage sensitivity in two nonmodel fish species, sterlet (Acipenser ruthenus) and common carp (Cyprinus carpio). Embryos of these species were exposed to a model genotoxicant, camptothecin (CPT), during cleavage (2-cell) stage and gastrulation. Results revealed a species-specific DNA damage sensitivity only at cleavage stage. 3 nM CPT caused lethality in sterlet embryos while carp embryos hatched normally. Multiple nuclear abnormalities were observed in sterlet embryos by early gastrula stage. However, carp embryos exhibited nuclear abnormalities and DNA fragmentation at neurula stage only when exposed to 7 nM CPT. Moreover, increased expression of tp53 in carp embryos at gastrula stage suggests activation of apoptosis mechanism. These findings suggest that carp embryos activate DNA damage response more efficiently than sterlet embryos at same developmental stage.

• Klíčová slova
• DNA repair, Development, Fish, Nuclear abnormality, Species-specific,
• MeSH
• apoptóza účinky léků   MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• druhová specificita * MeSH
• embryo nesavčí * účinky léků   MeSH
• fragmentace DNA MeSH
• kamptothecin * toxicita   analogy a deriváty   MeSH
• kapři * embryologie   genetika   MeSH
• mutageny toxicita   MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• poškození DNA * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• kamptothecin * MeSH
• mutageny MeSH
• nádorový supresorový protein p53 MeSH