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Temozolomide and sorafenib as programmed cell death inducers of human glioma cells
Joanna Jakubowicz-Gil1, Dorota Bądziul2, Ewa Langner3
1Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Lublin, Poland.
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.
Insights
Sorafenib and Temozolomide combination therapy effectively eliminates human glioma cells by inducing apoptosis and autophagy. Blocking heat shock proteins enhances sensitivity to endoplasmic reticulum stress and apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Gliomas are aggressive brain tumors known for high resistance to chemotherapy.
- Investigating novel therapeutic strategies is crucial for improving patient outcomes.
Purpose of the Study:
- To evaluate the combined effectiveness of sorafenib and Temozolomide in eliminating human glioma cells.
- To elucidate the roles of apoptosis and autophagy in drug-induced cell death.
Main Methods:
- Utilized MOGGCCM and T98G human glioma cell lines.
- Assessed cell morphology, apoptosis, autophagy, and mitochondrial membrane potential via microscopy and flow cytometry.
- Analyzed protein levels (Beclin1, LC3, p62, Hsps) and caspase activity.
- Investigated the impact of heat shock protein (Hsp) inhibition using siRNA.
Main Results:
- Temozolomide primarily induced autophagy in MOGGCCM cells, while sorafenib induced apoptosis.
- The drug combination synergistically enhanced apoptosis in a caspase-independent manner in MOGGCCM cells.
- In T98G cells, sorafenib induced autophagy and Temozolomide induced apoptosis; their combination led to dominant apoptosis.
- Hsp27 and Hsp72 inhibition increased sensitivity to ER stress and apoptosis induction.
Conclusions:
- Combined sorafenib and Temozolomide are potent inducers of apoptosis in glioma cells.
- Endoplasmic reticulum stress precedes glioma cell elimination.
- Hsp expression blocking impacts ER stress more significantly than apoptosis induction.
