The effects of chitosan-loaded JQ1 nanoparticles on OVCAR-3 cell cycle and apoptosis-related gene expression
Ehsan Masoudi1, Mitra Soleimani1, Giti Zarinfard1
1Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Background And Purpose:
Ovarian cancer is the deadliest gynecological cancer. Bromodomain and extra terminal domain (BET) proteins play major roles in the regulation of gene expression at the epigenetic level. Jun Qi (JQ1) is a potent inhibitor of BET proteins. Regarding the short half-life and poor pharmacokinetic profile, JQ1 was loaded into newly developed nano-carriers. Chitosan nanoparticles are one of the best and potential polymers in cancer treatment. The present study aimed to build chitosan-JQl nanoparticles (Ch-J-NPs), treat OVCAR-3 cells with Ch-J-NPs, and evaluate the effects of these nanoparticles on cell cycle and apoptosis-associated genes.
Experimental Approach:
Ch-J-NPs were synthesized and characterized. The size and morphology of Ch-J-NPs were defined by DLS and FE-SEM techniques. OVCAR-3 cells were cultured and treated with Ch-J-NPs. Then, IC50 was measured using MTT assay. The groups were defined and cells were treated with IC50 concentration of Ch-J-NPs, for 48 h. Finally, cells in different groups were assessed for the expression of genes of interest using quantitative RT-PCR.
Findings/Results:
IC50 values for Ch-J-NPs were 5.625 μg/mL. RT-PCR results demonstrated that the expression of genes associated with cell cycle activity (c-MYC, hTERT, CDK1, CDK4, and CDK6) was significantly decreased following treatment of cancer cells with Ch-J-NPs. Conversely, the expression of caspase-3, and caspase-9 significantly increased. BAX (pro-apoptotic) to BCL2 (anti-apoptotic) expression ratio, also increased significantly after treatment of cells with Ch-J-NPs.
Conclusion And Implications:
Ch-J-NPs showed significant anti-cell cyclic and apoptotic effects on OVCAR-3 cells.
Insights
Chitosan-Jun Qi nanoparticles (Ch-J-NPs) effectively target ovarian cancer cells. These nanoparticles inhibit cell cycle progression and induce apoptosis by modulating key gene expression, offering a promising therapeutic strategy.
Area of Science:
- Nanotechnology in Medicine
- Epigenetics and Cancer Therapeutics
- Gynecologic Oncology
Background:
- Ovarian cancer, the deadliest gynecological malignancy, necessitates novel therapeutic approaches.
- Bromodomain and extra terminal domain (BET) proteins regulate gene expression epigenetically; Jun Qi (JQ1) inhibits BET proteins.
- Chitosan nanoparticles offer a viable drug delivery system to overcome JQ1's poor pharmacokinetic profile.
Purpose of the Study:
- To synthesize and characterize chitosan-JQ1 nanoparticles (Ch-J-NPs).
- To evaluate the anti-cancer effects of Ch-J-NPs on OVCAR-3 ovarian cancer cells.
- To assess the impact of Ch-J-NPs on cell cycle and apoptosis-associated gene expression.
Main Methods:
- Ch-J-NPs were synthesized and characterized for size and morphology using DLS and FE-SEM.
- OVCAR-3 cells were treated with varying concentrations of Ch-J-NPs, with IC50 determined via MTT assay.
- Quantitative RT-PCR was employed to analyze the expression of cell cycle and apoptosis-related genes post-treatment.
Main Results:
- The IC50 of Ch-J-NPs was determined to be 5.625 μg/mL.
- Ch-J-NPs significantly downregulated genes involved in cell cycle progression (c-MYC, hTERT, CDK1, CDK4, CDK6).
- Treatment with Ch-J-NPs significantly upregulated pro-apoptotic genes (caspase-3, caspase-9) and increased the BAX/BCL2 ratio.
Conclusions:
- Ch-J-NPs demonstrate significant anti-proliferative effects on OVCAR-3 ovarian cancer cells.
- Ch-J-NPs effectively induce apoptosis in ovarian cancer cells.
- These findings highlight the potential of Ch-J-NPs as a novel therapeutic agent for ovarian cancer.
