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Radiation-Induced Autophagy Contributes to Cell Death and Induces Apoptosis Partly in Malignant Glioma Cells / Journal of the Korean Cancer Association, 대한암학회지
Cancer Research and Treatment ; : 221-241, 2015.
Article in English | WPRIM | ID: wpr-126956
ABSTRACT

PURPOSE:

Radiation-induced autophagy has been shown to play two different roles, in malignant glioma (MG) cells, cytocidal or cytoprotective. However, neither the role of radiation-induced autophagy for cell death nor the existence of autophagy-induced apoptosis, a well-known cell-death pathway after irradiation, has been verified yet. MATERIALS AND

METHODS:

We observed both temporal and dose-dependent response patterns of autophagy and apoptosis to radiation in MG cell lines. Additionally, we investigated the role of autophagy in apoptosis through knockdown of autophagy-related proteins.

RESULTS:

Autophagic activity measured by staining of acidic vesicle organelles and Western blotting of LC-3 protein increased in proportion to radiation dose from day 1 to 5 after irradiation. Apoptosis measured by annexin-V staining and Western blotting of cleaved poly(ADP-ribose) polymerase demonstrated relatively late appearance 3 days after irradiation that increased for up to 7 days. Blocking of pan-caspase (Z-VAD-FMK) did not affect apoptosis after irradiation, but silencing of Atg5 effectively reduced radiation-induced autophagy, which decreased apoptosis significantly. Inhibition of autophagy in Atg5 knockdown cells was shown to be beneficial for cell survival. Stable transfection of GFP-LC3 cells was observed after irradiation. Annexin-V was localized in cells bearing GFP-LC3 punctuated spots, indicating autophagy in immunofluorescence. Some of these punctuated GFP-LC3 bearing cells formed conglomerated spots and died in final phase.

CONCLUSION:

These findings suggest that autophagy appears earlier than apoptosis after irradiation and that a portion of the apoptotic population that appears later is autophagy-dependent. Thus, autophagy is a pathway to cell death after irradiation of MG cells.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Autophagy / Transfection / Organelles / Cell Line / Cell Survival / Blotting, Western / Fluorescent Antibody Technique / Cell Death / Poly(ADP-ribose) Polymerases / Apoptosis Language: English Journal: Cancer Research and Treatment Year: 2015 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Autophagy / Transfection / Organelles / Cell Line / Cell Survival / Blotting, Western / Fluorescent Antibody Technique / Cell Death / Poly(ADP-ribose) Polymerases / Apoptosis Language: English Journal: Cancer Research and Treatment Year: 2015 Type: Article