Photodynamic Therapy in Combination with Talaporfin Sodium Induces Mitochondrial Apoptotic Cell Death Accompanied with Necrosis in Glioma Cells

Miki, Yuichi; Akimoto, Jiro; Yokoyama, Sakino; Homma, Tomomi; Tsutsumi, Masateru; Haraoka, Jo; Hirano, Kazuya; Beppu, Masatoshi

doi:10.1248/bpb.b12-00567

Cited by 39 publications

(30 citation statements)

References 29 publications

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“…As we have previously shown, activation of mTOR downstream effectors was reduced after PDT, suggesting a down-regulation of the mTOR complex activity and a subsequent possible rise of the autophagic flux [14]. On the other hand, PDT is known to induce both necrosis and apoptosis in glioblastoma [19,20]. Recently, Receptor Interacting Protein kinase 3 (RIP3), a key protein of the programmed necrosis pathway (necroptosis) was demonstrated to be involved in PDT-induced glioblastoma cell death induced by PDT [21][22][23] and other cell death inducers [24][25][26].…”

Section: Introductionmentioning

confidence: 66%

RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death

Fettweis¹,

Valentin

L’homme³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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Glioblastomas are the deadliest type of brain cancer and are frequently associated with poor prognosis and a high degree of recurrence despite removal by surgical resection and treatment by chemo-and radio-therapy. Photodynamic therapy (PDT) is a treatment well known to induce mainly necrotic and apoptotic cell death in solid tumors. 5-Aminolevulinic acid (5-ALA)-based PDT was recently shown to sensitize human glioblastoma cells (LN-18) to a RIP3 (Receptor Interacting Protein 3)-dependent cell death which is counter-acted by activation of autophagy. These promising results led us to investigate the pathways involved in cell death and survival mechanisms occurring in glioblastoma following PDT. In the present study, we describe a new TSC2 (Tuberous Sclerosis 2)-dependent survival pathway implicating MK2 (MAPKAPK2) kinase and 14-3-3 proteins which conducts to the activation of a pro-survival autophagy. Moreover, we characterized a new RIP3/TSC2 complex where RIP3 is suggested to promote cell death by targeting TSC2-dependent survival pathway. These results highlight (i) a new role of TSC2 to protect glioblastoma against PDT-induced cell death and (ii) TSC2 and 14-3-3 as new RIP3 partners.

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Section: Introductionmentioning

confidence: 66%

RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death

Fettweis¹,

Valentin

L’homme³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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“…Twenty-four hours after NPe6-PDT treatment, T98G cells were lysed with lysis buffer [15], and cell lysate containing 1 μg of protein was analyzed by SDS-PAGE under reducing conditions. The gel was transblotted to a polyvinylidenefluoride membrane and incubated with 5 % skim milk in DPBS(-) for blocking.…”

Section: Detection Of Lc3-i and Lc3-ii Proteins In T98g Cellsmentioning

confidence: 99%

“…Combination of NPe6-PDT with surgical resection was found to achieve better therapeutic results than conventional protocols, particularly in patients with newly diagnosed malignant glioma [14]. We also examined the effect of NPe6-PDT on cell death modalities in glioma cells, with findings showing that NPe6-PDT induces mitochondrial apoptotic cell death accompanied by necrotic cell death [15]. However, despite the involvement of necrosis in glioma cell death following NPe6-PDT [15], the necrotic mechanism remains poorly understood.…”

Section: Introductionmentioning

confidence: 99%

“…We also examined the effect of NPe6-PDT on cell death modalities in glioma cells, with findings showing that NPe6-PDT induces mitochondrial apoptotic cell death accompanied by necrotic cell death [15]. However, despite the involvement of necrosis in glioma cell death following NPe6-PDT [15], the necrotic mechanism remains poorly understood.…”

Section: Introductionmentioning

confidence: 99%

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Photodynamic therapy using talaporfin sodium induces concentration-dependent programmed necroptosis in human glioblastoma T98G cells

et al. 2015

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Photodynamic therapy (PDT) using photosensitizer induces several types of cell death, such as apoptosis, necrosis, and autophagy, depending on the PDT procedure, photosensitizer type, and cell type. We previously demonstrated that PDT using the photosensitizer talaporfin sodium (mono-L-aspartyl chlorine e6, NPe6; NPe6-PDT) induces both mitochondrial apoptotic and necrotic cell death in human glioblastoma T98G cells. However, details regarding the mechanism of necrosis caused by NPe6-PDT are unclear. Here, we investigated whether or not necroptosis, a recently suggested form of programmed necrosis, is involved in the necrotic cell death of NPe6-PDT-treated T98G cells. Leakage of lactate dehydrogenase (LDH) from the cell layer into conditioned medium was significantly increased by NPe6 (25 and 50 μg/ml)-PDT, indicating that NPe6-PDT induces necrosis in these cells. NPe6 (25 μg/ml)-PDT treatment also induced conversion of microtubule-associated protein 1 light-chain 3 (LC3)-I into phosphatidylethanolamine-conjugated LC3-II accompanying autophagosome formation, indicators of autophagy; however, of note, NPe6 (50 μg/ml)-PDT did not induce such autophagic changes. In addition, both necrostatin-1 (a necroptosis inhibitor) and knockdown of necroptotic pathway-related proteins [e.g., receptor interacting serine-threonine kinase (RIP)-1, RIP-3, and mixed lineage kinase domain-like protein (MLKL)] inhibited leakage of LDH caused by NPe6 (25 μg/ml)-PDT. Taken together, the present findings revealed that NPe6-PDT-induced necrotic cell death is mediated in part by the necroptosis pathway in glioblastoma T98G cells.

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“…Adherent cells were then collected by detaching the cells from the bottom of the culture well through incubation with trypsin-EDTA solution (Sigma-Aldrich, St. Louis, MO, USA) for 2 min. Activity of caspase-3, cell surface-exposed phosphatidylserine and/or PI staining in glioma cells were measured in these collected cells using commercial assay kits (caspase-3: NucView TM 488 caspase-3 assay kit for live cells, exposed phosphatidylserine and PI: MEBCYTO apoptosis kit), as described previously ( Miki et al, 2013).…”

Section: Detection Of Apoptosis and Necrosismentioning

confidence: 99%

Effect of talaporfin sodium-mediated photodynamic therapy on cell death modalities in human glioblastoma T98G cells

et al. 2014

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-While photodynamic therapy (PDT) is an effective treatment for glioma, induction of apoptotic cell death of glioma cells is important for ensuring efficacy and safety of PDT treatment in glioma patients, as necrotic cell death can induce late appearance of obstacles in treatment. Here, we investigated the relationship between type of cell death and PDT treatment conditions involved in laser and photosensitizer dosage in human glioblastoma T98G cells. Photosensitizer talaporfin sodium-mediated PDT (NPe6-PDT) treatment induced laser and NPe6 dose-dependent cell death in T98G cells, whereas almost all cells pretreated with NPe6 at ≥ 30 μg/mL were killed by laser irradiation, regardless of laser dose. Morphological analysis showed that combination of high doses of NPe6 and laser irradiation changes the dominant cell death process from apoptosis to necrosis. Biochemical analysis (detection of caspase-3 activity and staining of cell surface-exposed phosphatidylserine) also showed that increasing laser dose changes the type of cell death from apoptotic to necrotic cell death after high-dose treatment with NPe6. Lactate dehydrogenase leakage assay demonstrated that a laser dose of 5 J/cm 2 induced less leakage than 30 J/cm 2 . Our results suggested that type of glioma cell death in NPe6-PDT changed with fluctuations in laser and NPe6 dose, and that combination of 30 μg/mL NPe6 with 5 J/cm 2 laser is the best treatment condition for inducing an increase in apoptotic cells while keeping rate of necrotic cell death low in this in vitro study.

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Photodynamic Therapy in Combination with Talaporfin Sodium Induces Mitochondrial Apoptotic Cell Death Accompanied with Necrosis in Glioma Cells

Cited by 39 publications

References 29 publications

RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death

RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death

Photodynamic therapy using talaporfin sodium induces concentration-dependent programmed necroptosis in human glioblastoma T98G cells

Effect of talaporfin sodium-mediated photodynamic therapy on cell death modalities in human glioblastoma T98G cells

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