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Biological activity of dihydroartemisinin in canine osteosarcoma cell lines
Kenji Hosoya1, Sridhar Murahari, Albert Laio
1Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
Objective:
To evaluate the biological activity of dihydroartemisinin on canine osteosarcoma cell lines in vitro.
Sample Population:
4 canine osteosarcoma cell lines.
Procedures:
Cell viability assays were performed on canine osteosarcoma cell lines OSCA2, OSCA16, OSCA50, and D17 after 24, 48, and 72 hours of treatment with dihydroartemisinin at concentrations of 0.1 to 100 microM. Apoptosis was assessed by use of an ELISA for free nuclosomal DNA fragmentation and by western blot analysis for cleavage of caspase 3. Cell cycle analysis was performed by use of staining with propidium iodide and flow cytometry. Detection of reactive oxygen species (ROS) was conducted in the D17 cell line by use of 6-carboxy-2',7'-dihydrofluorescein diacetate and flow cytometry.
Results:
The concentration of dihydroartemisinin required for 50% inhibition of cell viability (IC50) was achieved in all 4 canine osteosarcoma cell lines and ranged from 8.7 to 43.6 microM. Induction of apoptosis was evident as an increase in nucleosomal DNA fragmentation, cleavage of caspase 3, and an increase in the population in the sub G0/G1 phase of the cell cycle detected by flow cytometry. Exposure to dihydroartemisinin also resulted in a decrease in the G0/G1 population. Iron-dependent generation of ROS was detected in dihydroartemisinin-treated D17 cells; ROS generation increased in a dose-dependent manner.
Conclusions And Clinical Relevance:
Incubation with dihydroartemisinin resulted in biological activity against canine osteosarcoma cell lines, which included induction of apoptosis and arrest of the cell cycle. Clinical trials of dihydroartemisinin in dogs with osteosarcoma should be conducted.
Insights
Dihydroartemisinin demonstrated significant biological activity against canine osteosarcoma cell lines in vitro. This compound induced apoptosis and cell cycle arrest, suggesting potential for future clinical trials in dogs with osteosarcoma.
Area of Science:
- Veterinary Oncology
- Pharmacology
- Cell Biology
Background:
- Canine osteosarcoma is an aggressive bone cancer with limited treatment options.
- Dihydroartemisinin, an antimalarial drug, has shown potential anticancer properties in various studies.
Purpose of the Study:
- To investigate the in vitro biological activity of dihydroartemisinin against canine osteosarcoma cell lines.
- To assess dihydroartemisinin's effects on cell viability, apoptosis, cell cycle progression, and reactive oxygen species (ROS) generation.
Main Methods:
- Four canine osteosarcoma cell lines (OSCA2, OSCA16, OSCA50, D17) were treated with dihydroartemisinin at varying concentrations (0.1–100 µM).
- Cell viability was assessed using cell viability assays.
- Apoptosis was evaluated through DNA fragmentation assays and Western blot analysis of caspase 3.
- Cell cycle distribution and ROS generation were analyzed using flow cytometry.
Main Results:
- Dihydroartemisinin effectively inhibited cell viability in all tested cell lines, with IC50 values ranging from 8.7 to 43.6 µM.
- Apoptosis was induced, evidenced by increased DNA fragmentation and caspase 3 cleavage.
- Dihydroartemisinin treatment led to cell cycle arrest, with an increased sub-G0/G1 population and decreased G0/G1 population.
- Iron-dependent ROS generation was observed in a dose-dependent manner in D17 cells.
Conclusions:
- Dihydroartemisinin exhibits significant in vitro biological activity against canine osteosarcoma cell lines.
- The compound induces apoptosis and cell cycle arrest, mediated in part by ROS generation.
- Clinical trials evaluating dihydroartemisinin for canine osteosarcoma are warranted.
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