The role of actin in the apoptotic cell death of P19 embryonal carcinoma cells.

NERADIL, Jakub, Renata VESELSKÁ and Augustin SVOBODA. The role of actin in the apoptotic cell death of P19 embryonal carcinoma cells. International Journal of Oncology. vol. 27, No 4, p. 1013-1022. ISSN 1019-6439. 2005.

Basic information

• Original name: The role of actin in the apoptotic cell death of P19 embryonal carcinoma cells.
• Name in Czech: Úloha aktinu v apoptóze embryonálních karcinomových buněk linie P19.
• Authors: NERADIL, Jakub (203 Czech Republic, guarantor), Renata VESELSKÁ (203 Czech Republic) and Augustin SVOBODA (203 Czech Republic).
• Edition: International Journal of Oncology, 2005, 1019-6439.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 2.681
• RIV identification code: RIV/00216224:14110/05:00012573
• Organization unit: Faculty of Medicine
• UT WoS: 000232088700018
• Keywords in English: apoptosis; actin cytoskeleton; fractin; caspase-3; all-trans retinoic acid; cell differentiation
• Tags: actin cytoskeleton, all-trans retinoic acid, apoptosis, caspase-3, cell differentiation, fractin

Abstract

The P19 mouse embryonal carcinoma cell line was used as a model for a study of apoptosis accompanying differentiation induced by all-trans retinoic acid (ATRA). Apoptosis was detected both on the basis of morphological features (nuclear fragmentation, blebbing of plasma membrane, and formation of apoptotic bodies), and by using DNA electrophoresis and flow-cytometric measurement of DNA content. Actin cytoskeleton was studied both on morphological and submicroscopic levels. ATRA-treated cells manifested apoptosis-specific changes in the distribution of actin foremost in association with their entry into executive phase of apoptosis, when F-actin cables participated in cell disintegration into apoptotic bodies. Using immunogold labeling, actin was also identified in centers of fragmenting apoptotic nuclei, in the disintegration of which it is likely involved as well. At the same time, a cleavage of actin by active caspase-3 was proved, resulting in the emergence of 32kDa fragment termed fractin. Measurement of F-actin and fractin content using flow cytometry showed an unequivocal decrease of F-actin and synchronous increase of fractin in the apoptotic population as compared to non-treated cells. Therefore, our results proved both actin proteolysis and active involvement of specific actin structures in the final cell disintegration during apoptosis in the P19 cells.

Abstract (in Czech)

The P19 mouse embryonal carcinoma cell line was used as a model for a study of apoptosis accompanying differentiation induced by all-trans retinoic acid (ATRA). Apoptosis was detected both on the basis of morphological features (nuclear fragmentation, blebbing of plasma membrane, and formation of apoptotic bodies), and by using DNA electrophoresis and flow-cytometric measurement of DNA content. Actin cytoskeleton was studied both on morphological and submicroscopic levels. ATRA-treated cells manifested apoptosis-specific changes in the distribution of actin foremost in association with their entry into executive phase of apoptosis, when F-actin cables participated in cell disintegration into apoptotic bodies. Using immunogold labeling, actin was also identified in centers of fragmenting apoptotic nuclei, in the disintegration of which it is likely involved as well. At the same time, a cleavage of actin by active caspase-3 was proved, resulting in the emergence of 32kDa fragment termed fractin. Measurement of F-actin and fractin content using flow cytometry showed an unequivocal decrease of F-actin and synchronous increase of fractin in the apoptotic population as compared to non-treated cells. Therefore, our results proved both actin proteolysis and active involvement of specific actin structures in the final cell disintegration during apoptosis in the P19 cells.

Links

• GA304/00/0403, research and development project: Name: Změny cytoskeletu buněk nádorových linií v průběhu indukované apoptózy v závislosti na stádiu diferenciace.

Investor: Czech Science Foundation, Changes of the cytoskeleton in tumour cell lines during induced apoptosis in relation to different stages of differentiation