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SUBCELLULAR DAMAGE KINETICS WITHIN CO‐CULTIVATED WI38 and VA13‐TRANSFORMED WI38 HUMAN FIBROBLASTS FOLLOWING 5‐AMINOLEVULINIC ACID‐INDUCED PROTOPORPHYRIN IX FORMATION
Author(s) -
ÜBerriegler K. P.,
Banieghbal E.,
Krammer B.
Publication year - 1995
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1995.tb02408.x
Subject(s) - protoporphyrin ix , endoplasmic reticulum , chemistry , biophysics , protoporphyrin , organelle , photosensitizer , mitochondrion , kinetics , cell , fibroblast , nucleus , apoptosis , microbiology and biotechnology , photodynamic therapy , biochemistry , biology , photochemistry , in vitro , porphyrin , physics , organic chemistry , quantum mechanics
Abstract The generation of the photosensitizer protoporphyrin IX (PpIX) in cells can be induced by externally applied 5‐aminolevulinic acid (ALA), with that bypassing the feedback control mechanism. The aim of the present study was to investigate the onset of destructive changes in living cocultivated WI38 and VA13‐transformed WI38 human fibroblasts following ALA incubation, PpIX production and subsequent irradiation by white halogen light with a dose of 2.2 kJ/m 2 . Specific fluorescence markers such as 3,3′‐dihexyloxacarbocyanine iodide for endoplasmic reticulum (ER) staining and dihydrorhodamine for intact mitochondria mapping combined with a low light imaging system are a versatile and sensitive tool to examine the photoinduced destruction of organelles in living cells, while artifacts are minimized. Mitochondria as primary targets of PpIX undergo a condensation under irradiation and are finally destroyed. Photodynamic treatment induces further a significant decomposition of ER, although PpIX localization could not be determined. Initial destabilization and vesiculation of ER is followed by a porous network with large cisternae (indicating the breakdown of cell integrity and cell/nucleus membrane damage). Normal cocultivated lung fibroblasts showed a delay in destruction compared to the transformed WI38‐VA13 cells. The observed decomposition pattern resembles the morphological pattern of apoptosis.