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Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells In Vitro
Published on: March 18, 2014
Nanosecond pulsed electric field inhibits proliferation and induces apoptosis in human osteosarcoma
Xudong Miao1, Shengyong Yin2, Zhou Shao2
1The Department of Orthopedics, the Second Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang Province, 310003, China.
Objective:
Recent studies suggest that nanosecond pulsed electric field (nsPEF) is a novel minimal invasive and non-thermal ablation method that can induce apoptosis in different solid tumors. But the efficacy of nsPEF on bone-related tumors or bone metastasis is kept unknown. The current study investigates antitumor effect of nsPEF on osteosarcoma MG-63 cells in vitro.
Method:
MG-63 cells were treated with nsPEF with different electric field strengths (0, 10, 20, 30, 40, and 50 kV/cm) and different pulse numbers (0, 6, 12, 18, 24, and 30 pulses). The inhibitory effect of nsPEF on the growth of MG-63 cells was measured by Cell Counting Kit-8 (CCK-8) assay at different time points (0, 3, 12, 24, and 48 h post nsPEF treatment). The apoptosis was analyzed by Hoechst stain, in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL), and flow cytometric analysis. The expression of osteoprotegerin (OPG), receptor activator of NF-κB ligand (RANKL), and tumor necrosis factor α (TNF-α) was examined by reverse-transcription polymerase chain reaction (RT-PCR) and western blot.
Results:
The CCK-8 assay showed that nsPEF induced a distinct electric field strength- and pulse number-dependent reduction of cell proliferation. For treatment parameter optimizing, the condition 40 kV/cm and 30 pulses at 24 h post nsPEF achieved the most significant apoptotic induction rate. Hoechst, TUNEL, and flow cytometric analysis showed that the cell apoptosis was induced and cells were arrested in the G0/G1 phase. PCR and western blot analysis demonstrated that nsPEF up-regulated OPG expression had no effect on RANKL, increased OPG/RANKL ratio.
Conclusion:
NsPEF inhibits osteosarcoma growth, induces apoptosis, and affects bone metabolism by up-regulating OPG, indicating nsPEF-induced apoptosis in osteosarcoma MG-63 cells. NsPEF has potential to treat osteosarcoma or bone metastasis. When nsPEF is applied on metastatic bone tumors, it might be beneficial by inducing osteoblastic differentiation without cancer proliferation. In the future, nsPEF might be one of the treatments of metastatic bone tumor.
Insights
Nanosecond pulsed electric field (nsPEF) therapy effectively inhibits osteosarcoma cell growth and induces apoptosis. This novel treatment impacts bone metabolism by increasing osteoprotegerin (OPG) levels, showing potential for bone tumor treatment.
Area of Science:
- Oncology
- Biophysics
- Biotechnology
Background:
- Nanosecond pulsed electric field (nsPEF) is a promising non-thermal ablation technique for solid tumors.
- The efficacy of nsPEF on bone-related malignancies, including osteosarcoma, remains largely unexplored.
Purpose of the Study:
- To investigate the antitumor effects of nsPEF on osteosarcoma MG-63 cells in vitro.
- To determine the optimal nsPEF parameters for inducing apoptosis in osteosarcoma cells.
Main Methods:
- MG-63 cells were exposed to varying nsPEF strengths (0-50 kV/cm) and pulse numbers (0-30).
- Cell proliferation was assessed using CCK-8 assays; apoptosis was analyzed via Hoechst staining, TUNEL assays, and flow cytometry.
- Gene and protein expression of OPG, RANKL, and TNF-α were evaluated using RT-PCR and Western blot.
Main Results:
- nsPEF significantly reduced cell proliferation in a dose- and pulse-dependent manner.
- Optimal conditions (40 kV/cm, 30 pulses) induced significant apoptosis and G0/G1 cell cycle arrest.
- nsPEF upregulated OPG expression, increased the OPG/RANKL ratio, and had no effect on RANKL expression.
Conclusions:
- nsPEF effectively inhibits osteosarcoma cell growth and induces apoptosis.
- The treatment modulates bone metabolism markers, specifically upregulating OPG.
- nsPEF demonstrates potential as a therapeutic strategy for osteosarcoma and bone metastases, possibly by promoting osteoblastic differentiation.
