Joanna Jakubowicz-Gil1, Dorota Bądziul2, Ewa Langner3, Iwona Wertel4, Adrian Zając5, Wojciech Rzeski6. 1. Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Lublin, Poland. Electronic address: jjgil@poczta.umcs.lublin.pl. 2. Department of Human Physiology, University of Rzeszow, Rzeszów, Poland. Electronic address: dorota.badziul@wp.pl. 3. Department of Medical Biology, Institute of Agricultural Medicine, Lublin, Poland. Electronic address: ewa.langner@gmail.com. 4. 1st Department of Gynecology, University School of Medicine, Lublin, Poland. Electronic address: iwonawertel@wp.pl. 5. Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Lublin, Poland. Electronic address: a.zajac.umcs@gmail.com. 6. Department of Medical Biology, Institute of Agricultural Medicine, Lublin, Poland; Department of Immunology and Virology, Maria Curie-Sklodowska University, Lublin, Poland. Electronic address: rzeskiw@hektor.umcs.lublin.pl.
Abstract
BACKGROUND: Gliomas are aggressive brain tumors with very high resistance to chemotherapy. Therefore, the aim of the present study was to investigate the effectiveness of sorafenib and Temozolomide in elimination of human glioma cells through apoptosis and autophagy. METHODS: MOGGCCM (anaplastic astrocytoma) and T98G (glioblastoma multiforme) cell lines incubated with sorafenib and/or Temozolomide were used in the experiments. Cell morphology (ER stress, apoptosis, autophagy, and necrosis) was analyzed microscopically while apoptosis and mitochondrial membrane potential were assessed with flow cytometry. Beclin1, LC3, p62, Hsp27, and Hsp72 levels were analyzed by immunoblotting. The activity of caspase 3, 8, and 9 was evaluated fluorometrically. Expression of Hsps was blocked by transfection with specific siRNA. RESULTS: In MOGGCCM cells, Temozolomide most frequently induced autophagy, which was accompanied by decreased p62 and increased beclin1 and LC3II levels. Sorafenib initiated mainly apoptosis. Additional incubation with Temozolomide, synergistically potentiated the pro-apoptotic properties of sorafenib, but it was mediated in a caspase-independent way. In T98G cells, the effect of the analyzed drugs on programmed cell death induction was different from that in MOGGCCM cells. Sorafenib induced autophagy, while Temozolomide initiated mainly apoptosis. After simultaneous drug application, apoptosis dominated, suggesting synergistic action of both drugs. Inhibition of Hsp27 and Hsp72 expression increased the sensitivity of both cell lines to ER stress and, to a lesser extent, to induction of apoptosis, but not autophagy. CONCLUSIONS: Sorafenib and Temozolomide applied in combination are potent apoptosis inducers in T98G and MOGGCCM cells. ER stress precedes the elimination. Blocking of Hsp expression has a greater impact on ER stress rather than apoptosis induction.
BACKGROUND:Gliomas are aggressive brain tumors with very high resistance to chemotherapy. Therefore, the aim of the present study was to investigate the effectiveness of sorafenib and Temozolomide in elimination of humanglioma cells through apoptosis and autophagy. METHODS: MOGGCCM (anaplastic astrocytoma) and T98G (glioblastoma multiforme) cell lines incubated with sorafenib and/or Temozolomide were used in the experiments. Cell morphology (ER stress, apoptosis, autophagy, and necrosis) was analyzed microscopically while apoptosis and mitochondrial membrane potential were assessed with flow cytometry. Beclin1, LC3, p62, Hsp27, and Hsp72 levels were analyzed by immunoblotting. The activity of caspase 3, 8, and 9 was evaluated fluorometrically. Expression of Hsps was blocked by transfection with specific siRNA. RESULTS: In MOGGCCM cells, Temozolomide most frequently induced autophagy, which was accompanied by decreased p62 and increased beclin1 and LC3II levels. Sorafenib initiated mainly apoptosis. Additional incubation with Temozolomide, synergistically potentiated the pro-apoptotic properties of sorafenib, but it was mediated in a caspase-independent way. In T98G cells, the effect of the analyzed drugs on programmed cell death induction was different from that in MOGGCCM cells. Sorafenib induced autophagy, while Temozolomide initiated mainly apoptosis. After simultaneous drug application, apoptosis dominated, suggesting synergistic action of both drugs. Inhibition of Hsp27 and Hsp72 expression increased the sensitivity of both cell lines to ER stress and, to a lesser extent, to induction of apoptosis, but not autophagy. CONCLUSIONS:Sorafenib and Temozolomide applied in combination are potent apoptosis inducers in T98G and MOGGCCM cells. ER stress precedes the elimination. Blocking of Hsp expression has a greater impact on ER stress rather than apoptosis induction.
Authors: Adel Rezaei Moghadam; Simone C da Silva Rosa; Ehsan Samiei; Javad Alizadeh; Jared Field; Philip Kawalec; James Thliveris; Mohsen Akbari; Saeid Ghavami; Joseph W Gordon Journal: Cell Death Discov Date: 2018-10-25
Authors: Víctor M Pérez-García; Luis E Ayala-Hernández; Juan Belmonte-Beitia; Philippe Schucht; Michael Murek; Andreas Raabe; Juan Sepúlveda Journal: PLoS Comput Biol Date: 2019-07-15 Impact factor: 4.475