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Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan localisation in cultured tumour cells.

Teiten MH, Bezdetnaya L, Morlière P, Santus R, Guillemin F - Br. J. Cancer (2003)

Bottom Line: Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution.Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan-mediated PDT in the MCF-7 cell line.To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan accumulation.

View Article: PubMed Central - PubMed

Affiliation: Unité de Recherche en Thérapie Photodynamique, Centre Alexis Vautrin, F-54511 Vandoeuvre-les-Nancy Cedex, France.

ABSTRACT
Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution. Foscan subcellular localisation in the MCF-7 human adenocarcinoma cell line has been studied by means of confocal microscopy and microspectrofluorometry. The fluorescence topographic profiles recorded after cells costained with Foscan and organelle-specific fluorescent probes revealed that Foscan presents low localisation in lysosomes and a weak accumulation in mitochondria. Alternatively, the Foscan fluorescence topographic profile turned out to colocalise perfectly with that obtained for the endoplasmic reticulum (ER) and the Golgi apparatus. The patterns of fluorescence derived from confocal microscopy studies were consistent with predominant localisation of Foscan in these organelles. Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan-mediated PDT in the MCF-7 cell line. To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan accumulation.

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Intracellular fluorescence spectra of Rh123 alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated for 3 h with 1 μg ml−1 Foscan® and then with 10 μM Rh123 for 45 min. The spectrum in (B) was recorded at position 13.5 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 545 nm (Rh123) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×63, exposure time 4 s.
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fig3: Intracellular fluorescence spectra of Rh123 alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated for 3 h with 1 μg ml−1 Foscan® and then with 10 μM Rh123 for 45 min. The spectrum in (B) was recorded at position 13.5 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 545 nm (Rh123) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×63, exposure time 4 s.

Mentions: Intracellular fluorescence spectra of LY alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated overnight with 125 μg ml−1 LY and Foscan® (1 μg ml−1, 3 h). The spectrum in (B) was recorded at position 16.6 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 550 nm (LY) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×40, exposure time 4 s.


Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan localisation in cultured tumour cells.

Teiten MH, Bezdetnaya L, Morlière P, Santus R, Guillemin F - Br. J. Cancer (2003)

Intracellular fluorescence spectra of Rh123 alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated for 3 h with 1 μg ml−1 Foscan® and then with 10 μM Rh123 for 45 min. The spectrum in (B) was recorded at position 13.5 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 545 nm (Rh123) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×63, exposure time 4 s.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376781&req=5

fig3: Intracellular fluorescence spectra of Rh123 alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated for 3 h with 1 μg ml−1 Foscan® and then with 10 μM Rh123 for 45 min. The spectrum in (B) was recorded at position 13.5 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 545 nm (Rh123) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×63, exposure time 4 s.
Mentions: Intracellular fluorescence spectra of LY alone (A). Intracellular fluorescence spectra (B) and fluorescence topographic profiles (C) of MCF-7 cells incubated overnight with 125 μg ml−1 LY and Foscan® (1 μg ml−1, 3 h). The spectrum in (B) was recorded at position 16.6 μm on the X-axis of the profile in (C). Fluorescence topographic profiles in (C) were recorded at 550 nm (LY) and at 654 nm (Foscan®). Excitation wavelength 435 nm, objective magnification×40, exposure time 4 s.

Bottom Line: Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution.Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan-mediated PDT in the MCF-7 cell line.To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan accumulation.

View Article: PubMed Central - PubMed

Affiliation: Unité de Recherche en Thérapie Photodynamique, Centre Alexis Vautrin, F-54511 Vandoeuvre-les-Nancy Cedex, France.

ABSTRACT
Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution. Foscan subcellular localisation in the MCF-7 human adenocarcinoma cell line has been studied by means of confocal microscopy and microspectrofluorometry. The fluorescence topographic profiles recorded after cells costained with Foscan and organelle-specific fluorescent probes revealed that Foscan presents low localisation in lysosomes and a weak accumulation in mitochondria. Alternatively, the Foscan fluorescence topographic profile turned out to colocalise perfectly with that obtained for the endoplasmic reticulum (ER) and the Golgi apparatus. The patterns of fluorescence derived from confocal microscopy studies were consistent with predominant localisation of Foscan in these organelles. Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan-mediated PDT in the MCF-7 cell line. To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan accumulation.

Show MeSH
Related in: MedlinePlus