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Optimization of Western blotting analysis for the isolation and detection of membrane xenobiotic transporter ABCG2

100%

Szczygieł M., Markiewicz M., Szafraniec M., Zuziak R., Urbańska K., Fiedor L.

2017

vol. 64

issue 3

437-443

All organisms are exposed to numerous stress factors, which include harmful xenobiotics. The diversity of these compounds is enormous, thus in the course of evolution diverse biological defense mechanisms at various levels of organization have developed. One of them engages an evolutionarily conserved family of transporters from the ABC superfamily, found in most species - from bacteria to humans. An important example of such a transporter is the breast cancer resistance protein (BCRP/ABCG2), a typical integral membrane protein. It plays a key role in the absorption, distribution and elimination of a wide variety of xenobiotics, including drugs used in chemotherapy, and is involved in multidrug resistance. It also protects against phototoxic chlorophyll derivatives of dietary origin. BCRP is a hemitransporter which consists of one transmembrane domain, made of six alpha-helices forming a characteristic pore structure, and one ATP-binding domain, which provides the energy from ATP hydrolysis, required for active transport of the substrates. The isolation of BCRP is still not an easy task, because its insolubility in water and the presence of membrane rafts pose serious methodological and technical challenges during the purification. The aim of this study was to optimize the methods for detection and isolation of BCRP-enriched fractions obtained from animal tissue samples. In this report we describe an optimization of isolation of a BCRP-enriched membrane fraction, which is suitable for further protein quantitative and qualitative analysis using the molecular biology tools.

Tritolylporphyrin dimer as a new potent hydrophobic sensitizer for photodynamic therapy of melanoma.

84%

Drzewiecka A., Urbańska K., Matuszak Z., Pineiro M., Arnaut L. G., Habdas J., Ratuszna A., Stochel G.

2001

vol. 48

issue 1

277-282

We report the synthesis, photochemical and photophysical properties and preliminary studies on biological effect of a new tritolylporphyrin dimer (T-D). Absorption and emission properties of T-D suggest its possible use in photodynamic therapy. T-D is capable of singlet oxygen production with 0.8 quantum yield. It also has a high photostability. The photodynamic properties of the dimer were examined following the growth of SKMEL 188 (human melanoma) cells irradiated with red light (cut off <630 nm). The surviving fraction of the cells decreased about 3-fold (vs. non-irradiated cells) for an 81 J/cm2 dose. Our results suggest that tritolylporphyrine dimer T-D may be an interesting hydrophobic sensitizer for photodynamic therapy.

Metastasis inhibition after proton beam, β- and γ-irradiation of melanoma growing in the hamster eye

84%

Romanowska-Dixon B., Elas M., Swakoń J., Sowa U., Ptaszkiewicz M., Szczygieł M., Krzykawska M., Olko P., Urbańska K.

2013

vol. 60

issue 3

307-311

Standard ocular tumor treatment includes brachytherapy, as well as proton therapy, particularly for large melanoma tumors. However, the effects of different radiation types on the metastatic spread is not clear. We aimed at comparing ruthenium (106Ru, emitting β electrons) and iodine (125I, γ-radiation) brachytherapy and proton beam therapy of melanoma implanted into the hamster eye on development of spontaneous lung metastases. Tumors of Bomirski Hamster Melanoma (BHM) implanted into the anterior chamber of the hamster eye grew aggressively and completely filled the anterior chamber within 8-10 days. Metastases, mainly in the lung, were found in 100% of untreated animals 30 days after enucleation. Tumors were irradiated at a dose of 3-10 Gy with a 106Ru plaque and at a dose of 6-14 Gy using a 125I plaque. The protons were accelerated using the AIC-144 isochronous cyclotron operating at 60 MeV. BHM tumors located in the anterior chamber of the eye were irradiated with 10 Gy, for the depth of 3.88 mm. All radiation types caused inhibition of tumor growth by about 10 days. An increase in the number of metastases was observed for 3 Gy of β-irradiation, whereas at 10 Gy an inhibition of metastasis was found. γ-radiation reduced the metastatic mass at all applied doses, and proton beam therapy at 10 Gy also inhibited the metastastic spread. These results are discussed in the context of recent clinical and molecular data on radiation effects on metastasis.

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3 Acta Biochimica Polonica

1 2017

1 2013

1 2001

3 Urbańska K.

2 Szczygieł M.

1 Arnaut L. G.

1 Drzewiecka A.

1 Elas M.

1 Fiedor L.

1 Habdas J.

1 Krzykawska M.

1 Markiewicz M.

1 Matuszak Z.

1 Olko P.

1 Pineiro M.

1 Ptaszkiewicz M.

1 Ratuszna A.

1 Romanowska-Dixon B.

1 Sowa U.

1 Stochel G.

1 Swakoń J.

1 Szafraniec M.

1 Zuziak R.

Search results

Optimization of Western blotting analysis for the isolation and detection of membrane xenobiotic transporter ABCG2

Tritolylporphyrin dimer as a new potent hydrophobic sensitizer for photodynamic therapy of melanoma.

Metastasis inhibition after proton beam, β- and γ-irradiation of melanoma growing in the hamster eye