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Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells
Published on: June 16, 2019
P53 status, and G2/M cell cycle arrest, are determining factors in cell-death induction mediated by ELF-EMF in
Romina Mehdizadeh1, Alireza Madjid Ansari2, Flora Forouzesh1
1Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Abstract:
The average survival of patients with glioblastoma is 12-15 months. Therefore, finding a new treatment method is important, especially in cases that show resistance to treatment. Extremely low-frequency electromagnetic fields (ELF-EMF) have characteristics and capabilities that can be proposed as a new cancer treatment method with low side effects. This research examines the antitumor effect of ELF-EMF on U87 and U251 glioblastoma cell lines. Flowcytometry determined the viability/apoptosis and distribution of cells in different phases of the cell cycle. The size of cells was assessed by TEM. Important cell cycle regulation genes mRNA expression levels were investigated by real-time PCR. ELF-EMF induced apoptosis in U87cells much more than U251 (15% against 2.43%) and increased G2/M cell population in U87 (2.56%, p value < 0.05), and S phase in U251 (2.4%) (data are normalized to their sham exposure). The size of U87 cells increased significantly after ELF-EMF exposure (overexpressing P53 in U251 cells increased the apoptosis induction by ELF-EMF). The expression level of P53, P21, and MDM2 increased and CCNB1 decreased in U87. Among the studied genes, MCM6 expression decreased in U251. Increasing expression of P53, P21 and decreasing CCNB1, induction of cell G2/M cycle arrest, and consequently increase in the cell size can be suggested as one of the main mechanisms of apoptosis induction by ELF-EMF; furthermore, our results demonstrate the possible footprint of P53 in the apoptosis induction by ELF-EMF, as U87 carry the wild type of P53 and U251 has the mutated form of this gene.
Insights
Extremely low-frequency electromagnetic fields (ELF-EMF) show promise as a novel glioblastoma treatment. ELF-EMF induced significant apoptosis and cell cycle arrest in U87 cells, highlighting its potential therapeutic effects.
Area of Science:
- Oncology
- Biophysics
Background:
- Glioblastoma (GBM) has a poor prognosis with average survival of 12-15 months.
- Treatment resistance necessitates novel therapeutic strategies for GBM.
- Extremely low-frequency electromagnetic fields (ELF-EMF) present a potential low-side-effect cancer treatment modality.
Purpose of the Study:
- To investigate the antitumor effects of ELF-EMF on glioblastoma cell lines (U87 and U251).
- To elucidate the mechanisms underlying ELF-EMF-induced cell death and cell cycle modulation.
Main Methods:
- Cell viability, apoptosis, and cell cycle distribution were assessed using flow cytometry.
- Cell size was evaluated using transmission electron microscopy (TEM).
- mRNA expression levels of key cell cycle regulatory genes were quantified via real-time PCR.
Main Results:
- ELF-EMF significantly increased apoptosis in U87 cells (15%) compared to U251 cells (2.43%).
- ELF-EMF induced G2/M cell cycle arrest in U87 cells and S phase accumulation in U251 cells.
- U87 cells exhibited increased size and overexpression of P53, P21, and MDM2, with decreased CCNB1 expression.
- U251 cells showed decreased MCM6 expression, and P53 overexpression enhanced ELF-EMF-induced apoptosis.
Conclusions:
- ELF-EMF exhibits significant antitumor effects on glioblastoma cells, primarily through apoptosis induction and cell cycle arrest.
- The p53 pathway appears to play a crucial role in mediating ELF-EMF's apoptotic effects, particularly in cells with wild-type p53.
- ELF-EMF represents a promising therapeutic avenue for glioblastoma, warranting further investigation.
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