38 research outputs found
Compositions of Anticancer Drug with Micellar Nanocarriers and Their Cytotoxicity
Asymmetric diblock (DBC) and triblock (TBC) copolymers contained biocompatible chemically complementary polyacrylamide and poly(ethylene oxide) (PAAm-b-PEO-b-PAAm) or its monomethyl ether (MEPEO-b-PAAm), and also partially hydrolyzed triblock copolymer derivative (TBChydr) were used to create micelles of a special type. The micelles obtained are characterized by small CMCs and large values of the Gibbs micellization energy, thus indicating a high stability of DBC, TBC and TBChydr micelles in aqueous solutions and the capabilities of their use to encapsulate and deliver poorly soluble and/or toxic drugs in living organism. Morphological features and size of DBC and TBC micelles were determined by TEM. The electron images demonstrated spherical micelles of a polymolecular type, monomolecular type and separate micelle aggregates. TBC and TBChydr micelles were used to examine in vitro anticancer activity of their compositions with doxorubicin (Dox). The created micelle systems showed the enhanced cytotoxicity as compared to individual Dox against murine leukemia cells of L1210 line, murine transformed fibroblasts of L929 line and human T-leukemia cells of Jurkat line and allow to achieve a high efficacy at low Dox concentrations (0,1÷3 µg·cm-3) that opens the great prospects for essential decrease in drug dose at chemotherapy
Cell Surface Glycans at SLE – Changes During Cells Death, Utilization for Disease Detection and Molecular Mechanism Underlying Their Modification
Visualization of melanoma tumor with lectin-conjugated rare-earth doped fluoride nanocrystals
Aim To develop specific fluorescent markers for melanoma
tumor visualization, which would provide high selectivity
and reversible binding pattern, by the use of carbohydrate-
recognizing proteins, lectins, combined with the
physical ability for imaging deep in the living tissues by utilizing
red and near infrared fluorescent properties of specific
rare-earth doped nanocrystals (NC).
Methods B10F16 melanoma cells were inoculated to
C57BL/6 mice for inducing experimental melanoma tumor.
Tumors were removed and analyzed by lectin-histochemistry
using LABA, PFA, PNA, HPA, SNA, GNA, and
NPL lectins and stained with hematoxylin and eosin. NPL
lectin was conjugated to fluorescent NaGdF4:Eu3+-COOH
nanoparticles (5 nm) via zero length cross-linking reaction,
and the conjugates were purified from unbound substances
and then used for further visualization of histological
samples. Fluorescent microscopy was used to visualize
NPL-NaGdF4:Eu3+ with the fluorescent emission at 600-720
nm range.
Results NPL lectin selectively recognized regions of undifferentiated
melanoblasts surrounding neoangiogenic foci
inside melanoma tumor, PNA lectin recognized differentiated
melanoblasts, and LCA and WGA were bound to tumor
stroma regions. NPL-NaGdF4:Eu3+ conjugated NC were
efficiently detecting newly formed regions of melanoma
tumor, confirmed by fluorescent microscopy in visible and
near infrared mode. These conjugates possessed high photostability
and were compatible with convenient xylenebased
mounting systems and preserved intensive fluorescent
signal at samples storage for at least 6 months.
Conclusion NPL lectin-NaGdF4:Eu3+ conjugated NC permitted
distinct identification of contours of the melanoma
tissue on histological sections using red excitation at 590-
610 nm and near infrared emission of 700-720 nm. These
data are of potential practical significance for development
of glycans-conjugated nanoparticles to be used for in vivo
visualization of melanoma tumor
Detection of novel autoantigens in patients with recurrent miscarriage: description of an approach and preliminary findings
Aim To develop and test a protocol for isolation of potential
auto-antigens from chorionic tissue that may be linked
to recurrent miscarriage (RM).
Methods The strategy included: 1) isolation of IgGs tightly
bound to chorionic tissue of RM patients by protein G
chromatography; 2) construction of affinity columns using
the chorionic antibodies for isolation of auto-antigens; 3)
enrichment of auto-antigens from detergent extracted solution
of chorionic proteins by affinity chromatography; 4)
separation by dodecyl sulfate-electrophoresis followed by
matrix-assisted laser desorption/ionization-time of flight
mass spectrometry identification.
Results Five potential auto-antigens were detected: neutral
alpha-glucosidase AB, endoplasmin, transitional endoplasmic
reticulum ATPase, putative endoplasmin-like protein,
and cytoplasmic actin 2.
Conclusions We developed a strategy for identification of
auto-antigens in the chorionic tissue of women with RM,
which could be of diagnostic and prognostic value
Identification of a 48kDa form of unconventional myosin 1c in blood serum of patients with autoimmune diseases
AbstractWe searched for protein markers present in blood serum of multiple sclerosis (MS), rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients in comparison to healthy human individuals. We used precipitation/extraction methods and MALDI TOF/TOF mass spectrometry, and identified a protein with Mr ~46kDa as a fragment of human unconventional myosin IC isoform b (Myo1C). Western blotting with specific anti-human Myo1C antibodies confirmed the identity. Screening of blood serum samples from different autoimmune patients for the presence of Myo1c revealed its high level in MS and RA patients, relatively low level in SLE patients, and undetected in healthy donors. These data are suggesting that the level of p46 Myo1C in blood serum is a potential marker for testing of autoimmune diseases
4-Thiazolidinone derivative Les-3833 effectively inhibits viability of human melanoma cells through activating apoptotic mechanisms
Aim To evaluate cytotoxic action of 4-thiazolidinone derivative
Les-3833 and study the mechanisms of its pro-apoptotic
action toward human melanoma cells and human tumor
cell lines of other tissue origin.
Methods The effect of Les-3833 or doxorubicin on the viability
of 9 cell lines was studied using MTT assay, while
human melanoma cells of WM793 line were additionally
examined using light and fluorescent microscopies for
evaluating cytomorphological changes. The Western-blot
and flow cytometric analyses were carried out to study signaling
pathways of melanoma cell cycling and death.
Results Les-3833 was the most efficient against melanoma
cells. Its half maximal inhibitory concentration (IC50)
was 0.22 μg/mL for WM793 cells and 0.3 μg/mL for SK-Mel-
28 melanoma cells. For human lung A549, breast MCF-7,
colon HCT116, and ovarian SKOV3 carcinoma cell lines IC50
was in between 2.5 to >5.0 μg/mL. Les-3833 was relatively
not toxic (IC50 > 5 μg/mL) for human embryonic kidney
HEK293 cells. Results of Annexin V/PI staining of melanoma
cells and activation of caspase 3, PARP, MAPK, and EndoG
protein suggest apoptosis in Les-3833-treated cells. Les-
3833 also induced ROS production in melanoma cells and
their arrest in G0/G1 phase of cell cycle.
Conclusion Novel 4-thiazolidinone derivative Les-3833 is
effective against human melanoma cells in vitro, and such
effect is tumor specific since it is much less pronounced in
human carcinoma and leukemia cells. In melanoma cells
Les-3833 induces apoptosis (morphological changes and
increased pro-apoptotic proteins), ROS production, and arrest
in G0/G1 phase of cell cycle
Antioxidants selenomethionine and D-pantethine decrease the negative side effects of doxorubicin in NL/Ly lymphoma-bearing mice
Aim To investigate the potential tissue-protective effects
of antioxidants selenomethionine and D-pantethine applied
together with doxorubicin (Dx) on NK/Ly lymphomabearing
mice. The impact of this chemotherapy scheme
on animal survival, blood cell profile, hepatotoxicity, glutathione
level, and activity of glutathione-converting enzymes
in the liver was compared with the action of Dx applied
alone.
Methods The hematological profile of animals was studied
by the analysis of blood smears under light microscopy.
Hepatotoxicity of studied drugs was evaluated measuring
the activity of alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) enzymes, De Ritis ratio, and coenzyme
A fractions by McDougal assay. Glutathione level in
animal tissues was measured with Ellman reagent, and the
activity of glutathione reductase, transferase, and peroxidase
was measured using standard biochemical assays.
Results D-pantethine (500 mg/kg) and, to a lower extent,
selenomethionine (600 μg/kg) partially reduced the negative
side effects (leukocytopenia and erythropenia) of Dx (5
mg/kg) in NK/Ly lymphoma bearing animals on the 14th
day of their treatment. This increased animal survival time
from 47-48 to 60+ days and improved the quality of their
life. This ability of D-pantethine and selenomethionine was
realized via hepatoprotective and immunomodulating activities.
D-pantethine also restored the levels of acid-soluble
and free CoA in the liver of tumor-bearing animals,
while selenomethionine caused the recovery of glutathione
peroxidase levels in the liver, which was significantly
diminished under Dx treatment. Both compounds decreased
glutathione level in the liver, which was considerably
induced by Dx.
Conclusions Antioxidants selenomethionine and D-pantethine
partially reversed the negative side effects of Dx in
NK/Ly lymphoma-bearing mice and significantly increased
the therapeutic efficiency of this drug in tumor treatment
Specific antioxidant compounds differentially modulate cytotoxic activity of doxorubicin and cisplatin: in vitro and in vivo study
Aim To use the antioxidant compounds (sodium selenite,
selenomethionine, D-pantethine) for modulation of cytotoxic
effect of doxorubicin and cisplatin toward wild type
and drug-resistant mutants of several human tumor cells.
Similar treatments were applied in vivo toward adult male
Wistar rats.
Methods Human tumor cells of different lines (HCT-116,
Jurkat and HL-60) with various mechanisms of drug-resistance
were treated with doxorubicin or cisplatin, alone or
in combination with sodium selenite, selenomethionine,
or D-pantethine. Cell viability, induction of apoptosis, and
production of O2
- radicals were measured. Activity of redox
potential modulating enzymes was measured in the liver
and blood plasma of adult male Wistar rats subjected to
similar treatments.
Results All antioxidants used in physiologically harmless
concentration inhibited cytotoxic action of doxorubicin
toward tumor cells sensitive to chemotherapy treatment
by 15%-30%, and slightly enhanced cytotoxic effect of this
medicine toward drug-resistant malignant cells. At the
same time, there was no significant effect of these antioxidants
on cisplatin action. Such effects were accompanied
by a complete inhibition of production of superoxide radicals
induced by doxorubicin. The results of in vivo study in
adult male Wistar rats were in agreement with the results
of in vitro study of human tumor cells.
Conclusion Protective effect of specific antioxidant agents
during cytotoxic action of doxorubicin was demonstrated
in vitro in drug-sensitive human tumor cells and in adult
male Wistar rats, while there was no protective effect in
drug-resistant sub-lines of these tumor cells during action
of doxorubicin and cisplatin
Luminescent SiO2 nanoparticles for cell labeling: combined water dispersion polymerization and 3D condensation controlled by oligoperoxide surfactant-initiator
Hybrid polymer coated silica nanoparticles (NPs) were synthesized using low temperature graft (co)polymerization of trimethoxysilane propyl methacrylate (MPTS) initiated by surface-active oligoperoxide metal complex (OMC) in aqueous media. These NPs were characterized by means of kinetic, solid-state NMR, TEM and FTIR techniques. Two processes, namely the radical graft-copolymerization due to presence of double bonds and 3D polycondensation provided by the intra- or/and intermolecular interaction of organosilicic fragments, occurred simultaneously. The relative contribution of the reactions depending on initiator concentration and pH value leading to the formation of low cured polydisperse microparticles or OMC coated SiO2 NPs of controlled curing degree was studied. The availability of free-radical forming peroxide fragments on the surface of SiO2 NPs provides an opportunity for seeded polymerization leading to the formation of the functional polymer coated NPs with controlled particle structure, size, and functionality. Encapsulation of the luminescent dye (Rhodamine 6G) in SiO2 core of functionalized NPs provided a noticeable increase in their resistance to photo-bleaching and improved biocompatibility. These luminescent NPs were not only attached to murine leukemia L1210 cells but also tolerated by the mammalian cells. Their potential use for labeling of the mammalian cells is considered
Landomycins as Glutathione-Depleting Agents and Natural Fluorescent Probes for Cellular Michael Adduct-Dependent Quinone Metabolism
Landomycins are angucyclines with promising antineoplastic activity produced by Streptomyces bacteria. The aglycone landomycinone is the distinctive core, while the oligosaccharide chain differs within derivatives. Herein, we report that landomycins spontaneously form Michael adducts with biothiols, including reduced cysteine and glutathione, both cell-free or intracellularly involving the benz[a]anthraquinone moiety of landomycinone. While landomycins generally do not display emissive properties, the respective Michael adducts exerted intense blue fluorescence in a glycosidic chain-dependent manner. This allowed label-free tracking of the short-lived nature of the mono-SH-adduct followed by oxygen-dependent evolution with addition of another SH-group. Accordingly, hypoxia distinctly stabilized the fluorescent mono-adduct. While extracellular adduct formation completely blocked the cytotoxic activity of landomycins, intracellularly it led to massively decreased reduced glutathione levels. Accordingly, landomycin E strongly synergized with glutathione-depleting agents like menadione but exerted reduced activity under hypoxia. Summarizing, landomycins represent natural glutathione-depleting agents and fluorescence probes for intracellular anthraquinone-based angucycline metabolism