|Reference : RIP3 Expression Induces a Death Proﬁle Change in U2OS Osteosarcoma Cells After 5-ALA-PDT|
|Scientific journals : Article|
|Life sciences : Biochemistry, biophysics & molecular biology|
|RIP3 Expression Induces a Death Proﬁle Change in U2OS Osteosarcoma Cells After 5-ALA-PDT|
|Coupienne, Isabelle [Université de Liège - ULg > > GIGA-R : Virologie - Immunologie >]|
|Fettweis, Grégory [Université de Liège - ULg > > > GIGA - Utilisateurs machines]|
|Piette, Jacques [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Virologie - Immunologie >]|
|Lasers in Surgery and Medicine|
|Wiley Liss, Inc.|
|[en] photodynamic therapy ; osteosarcoma ; RIP3|
|[en] Background and Objective
The receptor-interacting protein 3 (RIP3) has recently been outlined as a key necrosis mediator but is also thought to participate in the regulation of apoptosis. The aim of this study is to compare the cell death profile induced by 5-aminolevulic acid (5-ALA)-mediated photodynamic therapy (PDT) in the RIP3-deficient cell line U2OS and in U2OS cells in which the expression of RIP3 was restored.
Materials and Methods
RIP3-expressing U2OS cells (RIP3-U2OS) were obtained after transfection and antibiotic selection. Wild type and RIP3-U2OS cells were treated by 5-ALA-PDT. Overall cell viability was evaluated and different parameters characteristic of apoptosis, autophagy, and necrosis were studied.
Surprisingly, the survival of RIP3-U2OS cells was higher compared to that of the wild type cells. In addition, RIP3-U2OS cell death was decreased by a zVAD-fmk pre-treatment. A higher cleavage of caspase-3, 7, 8, 9, and PARP was also detected in these cells, pointing out to the activation of caspase-dependent apoptosis. In parallel, a thrust of autophagy was clearly identified in the RIP3-U2OS cells. Conversely, RIP3-U2OS exhibited a lower level of necrosis than the wild types. Interestingly, necrostatin-1 efficiently decreased necrosis level in RIP3-U2OS but not in wild type cells.
Expression of RIP3 in U2OS cells led to a better survival but also to a death profile change in response to PDT. The apoptotic and autophagic pathways were clearly up-regulated compared to the RIP3-deficient wild type cells. However, induction of necrosis was weaker in the RIP3-U2OS cells. In this context, autophagy is likely to play a protective role against PDT-induced cell death and to allow a better survival of RIP3-U2OS cells. This work also highlights the important role played by RIP3 in the apoptotic pathway, although the modalities are still widely unknown.
|Fonds pour la formation à la Recherche dans l'Industrie et dans l'Agriculture (Communauté française de Belgique) - FRIA ; Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS|
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