Photodynamic therapy is a very promising and constantly evolving diagnostic and therapeutic method that is used mainly for malignant and non-malignant tumors treatment. This study deals with the utilization of zinc photosensitizer (λmax ~ 660 nm) from the group of phthalocyanines in photodynamic therapy. The aim of this study is to evaluate in vitro effect of the 5 Jcm-2 zinc phthalocyanine photosensitizer-mediated photodynamic therapy in EC50 concentration (30 nM) on cytoskeletal apparatus of the tumor cell line-HeLa (cervical cancer cells). For the measurement, the tandem mass spectrometry, atomic force and fluorescent confocal microscopy techniques were used. The results showed, that compared to the control cells zinc-derivative mediated photodynamic therapy caused in HeLa cells significant change of the cell height and extensive cytoskeletal actin rearrangement although the levels of beta actin, gamma actin and F-actin did not change significantly. This is probably caused by decreased level of the ARPC2 actin-related protein which is responsible for actin polymerization. Its level decreased 24 hours after therapy by 56%. The cytoskeletal apparatus is one of the basic cellular structures that provides cell shape, cell division and the intracellular transport. After in vitro 5 Jcm(-2) zinc derivative-mediated photodynamic therapy, the cervical carcinoma cells showed a significant damage of the cytoskeletal structure followed by changes of cell shape leading to cell death. Considering these results and low effective concentration (EC50 = 30 nM), the therapy used is potentially very promising antitumor treatment.
Metalloporphyrins are an important group of sensitizers with a porphyrin skeleton. Their photophysical properties are significantly affected by the nature of the central ion. In this work, we focus on the mechanical properties of a cervix carcinoma cell line which underwent photodynamic treatment (PDT) with MgTPPS4 photosensitzer. Atomic force microscopy alongside confocal microscopy was used to quantify and qualify the structural characteristics before and after PDT. Cells before PDT showed a fine actin network and higher elasticity with the median of Young modulus 12.2 kPa. After PDT, the median of Young modulus was 13.4 kPa and a large redistribution in the actin network was observed.
- MeSH
- cytoskelet účinky léků fyziologie účinky záření MeSH
- fotosenzibilizující látky chemie farmakologie MeSH
- HeLa buňky MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- metaloporfyriny chemie farmakologie MeSH
- mikroskopie atomárních sil MeSH
- modul pružnosti MeSH
- světlo MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Oscillating polar entities inside the biological cells, most notably microtubules, are bound to emit electromagnetic radiation. This phenomenon is described by Fröhlich kinetic equations expressing, in terms of quantum occupancy numbers of each discrete collective oscillatory mode, the balance between incoming metabolic energy flow and losses due to linear and non-linear interactions with the thermal environs of the oscillators. Hitherto, radiation losses have not been introduced as part of the balance; it was assumed that they were proportional to the modal occupation numbers. It is demonstrated that this formulation is incorrect and the radiation losses must be taken into account in the kinetic equations explicitly. Results of a numerical study of kinetic equations, enlarged in this sense, are presented for the case of three coupled oscillators which was shown to evince the essential attributes of the Fröhlich systems. Oscillator eigenfrequencies were chosen, alternatively, to fall into the MHz and the THz frequency domains. It was found that large radiation levels destroy the main hallmark of the Fröhlich systems, the energy condensation in the lowest frequency mode. The system then functions as a convertor of metabolic energy into radiation. At more moderate radiation levels, both energy condensation and significant radiation can coexist. Possible consequences for the cell physiology are suggested.
- Klíčová slova
- PDT, detekce buněčného poškození, buněčná linie,
- MeSH
- biofyzika metody přístrojové vybavení výchova MeSH
- buněčné linie účinky záření MeSH
- cytoskelet účinky záření MeSH
- financování organizované MeSH
- fotochemoterapie metody přístrojové vybavení využití MeSH
- reaktivní formy kyslíku diagnostické užití MeSH
- školy lékařské MeSH
Coherent vibration states in biological systems excited in nonlinear electrically polar structures by metabolic energy supply were postulated by H. Frohlich. Frohlich's requirements for coherent vibrations and generation of electromagnetic field are satisfied by microtubules whose subunits are electric dipoles. Static electric field around mitochondria and "wasted energy" efflux from them provide nonlinear conditions and coherent excitation. Numerical models are used for analysis of coherent vibration states. A hypothesis is presented that dysfunction of mitochondria (i.e., extinction of the zones of the static electric field and of the efflux of "wasted energy") and disintegration of the cytoskeleton on the pathway of cancer transformation result in disturbances of coherence of the cellular electrically polar oscillations and of the generated electromagnetic field with consequences in cellular organization and interactions between cells. Local invasion, detachment, and metastasis of cancer cells are subsequent events of disturbed electromagnetic interactions.
- MeSH
- biofyzikální jevy MeSH
- biologické modely MeSH
- cytoskelet metabolismus účinky záření MeSH
- elektromagnetická pole škodlivé účinky MeSH
- lidé MeSH
- mitochondrie metabolismus účinky záření MeSH
- nádorová transformace buněk MeSH
- nádorové buněčné linie MeSH
- nádory patologie MeSH
- nelineární dynamika MeSH
- vibrace MeSH
- Check Tag
- lidé MeSH
K562 is the chronic myelogenous leukemia (CML)-derived cell line that expresses high levels of chimeric oncoprotein Bcr-Abl. The deregulated (permanent) kinase activity of Bcr-Abl leads to continuous proliferation of K562 cells and their resistance to the apoptosis promotion by conventional drugs. The photodynamic treatment (PDT) based on the application of 5-aminolevulinic acid (ALA) and irradiation with blue light (ALA-PDT) resulted in the suppression of K562 cells proliferation. It was followed by a necrosis-like cell death [K. Kuzelová, D. Grebenová, M. Pluskalová, I. Marinov, Z. Hrkal, J. Photochem. Photobiol. B 73 (2004) 67-78]. ALA-PDT led to the perturbation of the Hsp90/p23 multichaperone complex of which the Bcr-Abl is the client protein. Bcr-Abl protein was suppressed whereas the bcr-abl mRNA level was not affected. Further on, we observed several changes in the cytoskeleton organization. We detected ALA-PDT-mediated disruption of filamental actin structure using FITC-Phalloidin staining. In connection with this we uncovered certain cytoskeleton organizing proteins involved in the cell response to the treatment. Among these proteins, Septin2, which plays a role in maintaining actin bundles, was suppressed. Another one, PDZ-LIM domain protein 1 (CLP36) was altered. This protein acts as an adaptor molecule for LIM-kinase which phosphorylates and thus inactivates cofilin. Cofilin was indeed dephosphorylated and could thus be activated and operate as an actin-depolymerizing factor. We propose the scheme of molecular response of K562 cells to ALA-PDT.
- MeSH
- buněčná smrt účinky léků účinky záření MeSH
- buňky K562 patologie účinky léků účinky záření MeSH
- časové faktory MeSH
- cytoskelet patologie účinky léků účinky záření MeSH
- DNA vazebné proteiny metabolismus MeSH
- faloidin chemie MeSH
- financování organizované MeSH
- fluorescein-5-isothiokyanát MeSH
- fotosenzibilizující látky farmakologie MeSH
- kyselina aminolevulová farmakologie MeSH
- lidé MeSH
- Lim-kinasy MeSH
- messenger RNA metabolismus MeSH
- mikrofilamentové proteiny metabolismus MeSH
- molekulární chaperony metabolismus MeSH
- onkogenní proteiny metabolismus MeSH
- proteinkinasy fyziologie MeSH
- proteiny tepelného šoku HSP90 metabolismus MeSH
- regulace genové exprese MeSH
- světlo MeSH
- transkripční faktory MeSH
- transportní proteiny metabolismus MeSH
- tyrosinkinasy metabolismus účinky léků účinky záření MeSH
- Check Tag
- lidé MeSH
- MeSH
- cytoskelet účinky záření ultrastruktura MeSH
- lasery MeSH
- lidé MeSH
- nádorové buňky kultivované účinky záření ultrastruktura MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- techniky in vitro MeSH
