Chloroquine Enhances Mda-7- Induced Apoptosis via miR-7 and HSP70 Modulation in Glioblastoma
Seyedeh Maliheh Babazadeh1, Mohammad Reza Zolfaghari1, Mohsen Zargar1
1Department of Microbiology, Qo.C., Islamic Azad University, Qom, Iran.
Background:
Melanoma differentiation-associated gene-7 (Mda-7) selectively suppresses growth and induces apoptosis in various tumor cells without harming normal cells. Inhibition of autophagy has been shown to enhance the efficacy of many cancer therapies. However, its effect on the anticancer activity of Ad/Mda-7 in GBM has remained unclear. This study investigated the combined effect of an autophagy inhibitor (CQ) and Mda-7 in U87 cancer cells.
Methods:
Cell proliferation was assessed using the MTT assay. Apoptosis rates, autophagy induction, and ROS levels were measured using flow cytometry. Caspase-9 and β-actin protein levels were analyzed by Western blotting. ELISA was employed to quantify HSP70 and TRAIL level in the culture medium. Real-time PCR evaluated the expression levels of cell death-related genes (P38 MAPK, Bax, and TRAIL) and specific miRNAs (miR-7, miR-122, and miR-21) in treated cells.
Results:
Combined treatment with Ad/Mda-7 and the autophagy inhibitor CQ significantly reduced cell viability and proliferation. Ad/Mda-7 induced LC3-II protein expression in U87 cancer cells, which was further increased by autophagy inhibition through CQ. The combination treatment also increased apoptosis rates, elevated ROS levels, and decreased HSP70 protein expression, highlighting its synergistic anticancer effects. Increasing the expression of miR-7 and miR-122 indicated that the elevated levels of these endogenous miRNAs may help improve the treatment process.
Conclusion:
Our findings indicate that the combination of Ad/Mda-7 and CQ synergistically could inhibit U87 cancer cell growth and could serve as a promising approach for treating human GBM.
Insights
Combining Ad/Mda-7 with chloroquine (CQ) effectively inhibits U87 glioblastoma cell growth and proliferation. This synergistic approach enhances apoptosis and shows promise for treating human glioblastoma (GBM).
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Melanoma differentiation-associated gene-7 (Mda-7) exhibits selective anti-tumor properties.
- Autophagy inhibition can potentiate cancer therapies, but its effect with Ad/Mda-7 in glioblastoma (GBM) is unclear.
- This study examines the combined effects of Ad/Mda-7 and chloroquine (CQ) on U87 GBM cells.
Purpose of the Study:
- To investigate the synergistic anticancer effects of Ad/Mda-7 and CQ in U87 cancer cells.
- To evaluate the impact of combined treatment on cell viability, apoptosis, and autophagy.
- To explore the role of reactive oxygen species (ROS) and specific miRNAs in the treatment response.
Main Methods:
- Cell proliferation assessed via MTT assay.
- Apoptosis, autophagy, and ROS levels measured by flow cytometry.
- Protein analysis (Caspase-9, β-actin, HSP70) by Western blotting and ELISA.
- Gene and miRNA expression analyzed using real-time PCR.
Main Results:
- Combined Ad/Mda-7 and CQ treatment significantly reduced U87 cell viability and proliferation.
- Treatment induced apoptosis, elevated ROS levels, and decreased HSP70.
- Autophagy inhibition by CQ enhanced Ad/Mda-7's effects, increasing LC3-II expression.
- Upregulation of miR-7 and miR-122 suggests their involvement in the synergistic effect.
Conclusions:
- Ad/Mda-7 combined with CQ demonstrates synergistic inhibition of U87 cancer cell growth.
- This combination therapy presents a promising strategy for treating human glioblastoma (GBM).
- Further research into the role of specific miRNAs may optimize this therapeutic approach.
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