Combination therapy application of Abemaciclib with Doxorubicin in triple negative breast cancer cell line MDA-MB-231
Tugce Nur Eralp1, Akin Sevinc2, Banu Mansuroglu3
1Yildiz Technical University, Graduate School of Science and Engineering, Department of Molecular Biology and Genetics, Istanbul, 34220, Turkey. tugcenureralp@gmail.com.
Abstract:
Due to lack of clinical biomarkers, Triple Negative Breast Cancer (TNBC) is more likely to have spread to other tissues at time of diagnosis and therapy planning generally involves use of cytotoxic chemotherapy agents, such as Doxorubicin. We aimed to investigate possible advantages of using combination strategy using Doxorubicin alongside Abemaciclib. After determining the IC50 values for Doxorubicin (DOX) and Abemaciclib (ABE); CompuSyn and ComBenefit software were used to reveal the effect resulting from the combination of two drugs. Following the determined effect, cell death was revealed by fluorescence microscopy and a colony forming assay was performed to see the potential of even a single cancer cell with adhesive character to survive over time and form a clone of itself. Detection of changing antioxidant activity following DOX, ABE and DOX+ABE combination therapy in MDAMB231 cells was determined by measuring MDA, SOD and GSH activities. The expression of Cleaved Caspase 3, PARP, Cleaved PARP, Cdk2 and Bax, which changed as a result of DOX, ABE and DOX+ABE application, was shown by Western Blotting.Cyclin-dependent kinase inhibitors appear as promising agents in therapy planning for breast cancer due to their prominent role in cell cycle regulation, where the number of studies interrogating its efficiency in the treatment of cancer such as TNBC is limited. For this reason, in this study, we aimed to determine the impact of the combined use of the CDK4/6 inhibitors ABE and DOX on the cytotoxicity, apoptotic homeostasis, alterations in antioxidative mechanisms, and the molecular pathways that they utilize. Our results showed that when used in combination, Doxorubicin and Abemaciclib showed a synergistic effect on TNBC cell line MDA-MB-231.
Insights
Combining Doxorubicin with Abemaciclib, a cyclin-dependent kinase inhibitor, demonstrates a synergistic effect against Triple Negative Breast Cancer (TNBC). This combination therapy enhances cytotoxicity and induces apoptosis in TNBC cells, offering a promising strategy for treatment.
Area of Science:
- Oncology
- Pharmacology
- Molecular Biology
Background:
- Triple Negative Breast Cancer (TNBC) lacks clinical biomarkers, leading to advanced diagnoses and reliance on cytotoxic chemotherapy like Doxorubicin.
- Cyclin-dependent kinase (CDK) inhibitors are emerging as vital therapeutic agents for breast cancer due to their role in cell cycle regulation, yet their efficacy in TNBC requires further investigation.
Purpose of the Study:
- To investigate the potential advantages of combining Doxorubicin (DOX) with Abemaciclib (ABE), a CDK4/6 inhibitor, for TNBC treatment.
- To evaluate the synergistic effects of DOX and ABE on cytotoxicity, apoptosis, antioxidant activity, and molecular pathways in the MDA-MB-231 TNBC cell line.
Main Methods:
- Determined IC50 values for DOX and ABE, then utilized CompuSyn and ComBenefit software to analyze drug combination effects.
- Assessed drug impact on cell death via fluorescence microscopy and colony formation assays.
- Measured antioxidant activities (MDA, SOD, GSH) and protein expression (Cleaved Caspase 3, PARP, Cleaved PARP, Cdk2, Bax) using Western Blotting.
Main Results:
- The combination of Doxorubicin and Abemaciclib exhibited a synergistic effect against the MDA-MB-231 TNBC cell line.
- Combined therapy led to increased cytotoxicity and apoptosis, as evidenced by microscopy and colony formation assays.
- Significant alterations in antioxidant enzyme activities and key molecular markers associated with apoptosis and cell cycle regulation were observed.
Conclusions:
- The combination of Doxorubicin and Abemaciclib demonstrates significant synergistic efficacy in vitro against TNBC.
- This combination strategy holds promise for enhancing therapeutic outcomes in Triple Negative Breast Cancer treatment by targeting critical cellular pathways.
Related Concept Videos
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...
02:50Combination Therapies and Personalized Medicine
The combination of the drug acetazolamide and sulforaphane is a good example of combination therapy to treat cancer. The cells in the interior of a large tumor often die due to the hypoxic and...
02:34Inhibition of Cdk Activity