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High-throughput, Label-free Proteomics Identifies Salient Proteins and Genes in MDA-MB-231 Cells Treated with Natural
Gowrisree Varadarajan1, Jeya Shree Thulasidas1, Pragatheiswar Giri2
1Department of Electrical Engineering, Divison of High Voltage Engineering, Anna University, Guindy, India.
Abstract:
With the absence of the three most common receptor targets, and with high vascularity and higher-grade tumors, triple-negative breast cancer (TNBC) is the most aggressive of all breast cancer subtypes and is in need of additional/alternative/novel treatment strategies. With ~ 15% of the over 2 million new cases each year, there is an unmet need to treat TNBC. MDA-MB-231, human TNBC cells, were treated with neem leaf extract (Neem) and eight, 1200 V/cm, 100 µs electric pulses (EP), and their viability and proteomic profiles were studied. With EP + Neem, a lower viability of 37% was observed after 24 h, compared to 85% in the neem-only samples, indicating the efficacy of the combinational treatment. The proteomics results indicated significant upregulation of 525 proteins and downregulation of 572 proteins, with a number of different pathways in each case. These include a diverse group of proteins, such as receptors, heat shock proteins, and many others. The upregulated TCA cycle and OXPHOS pathways and the downregulated DNA replication and ubiquitin-mediated proteolytic pathways were associated with effective cell death, demonstrating the potency of this treatment. Viability results reveal the efficacious anticancer effects of the EP + Neem combination, via growth inhibition, on TNBC cells. Proteomics studies could readily identify the effected protein pathways, and their corresponding genes, that are responsible for cell death. This represents a potential therapeutic strategy against TNBC when patients are refractory to standard treatments.
Insights
Triple-negative breast cancer (TNBC) treatment combining neem leaf extract and electric pulses (EP+Neem) significantly reduced cancer cell viability. This novel approach offers a promising therapeutic strategy for TNBC patients resistant to standard treatments.
Area of Science:
- Oncology
- Biochemistry
- Molecular Biology
Background:
- Triple-negative breast cancer (TNBC) is an aggressive subtype lacking common receptor targets, necessitating novel treatment strategies.
- Approximately 15% of over 2 million annual breast cancer cases are TNBC, highlighting a significant unmet clinical need.
- Current treatment options for TNBC are limited, especially for patients refractory to standard therapies.
Purpose of the Study:
- To investigate the efficacy of a combination therapy using neem leaf extract (Neem) and electric pulses (EP) against human TNBC cells (MDA-MB-231).
- To analyze the impact of this combination treatment on cell viability and proteomic profiles.
- To identify molecular pathways affected by the combined therapy that contribute to cancer cell death.
Main Methods:
- Human TNBC cells (MDA-MB-231) were treated with neem leaf extract and electric pulses (1200 V/cm, 100 µs).
- Cell viability was assessed after 24 hours to evaluate treatment efficacy.
- Proteomic profiling was performed to identify differentially expressed proteins and affected cellular pathways.
Main Results:
- The combination of EP + Neem resulted in significantly lower cell viability (37%) compared to Neem alone (85%) after 24 hours.
- Proteomics revealed significant upregulation of 525 proteins and downregulation of 572 proteins.
- Upregulated pathways included the TCA cycle and OXPHOS, while downregulated pathways included DNA replication and ubiquitin-mediated proteolysis, correlating with effective cell death.
Conclusions:
- The EP + Neem combination demonstrates potent anticancer effects, inhibiting growth and inducing cell death in TNBC cells.
- Proteomic analysis successfully identified key protein pathways and genes involved in mediating the therapeutic effects.
- This combination therapy represents a potential alternative treatment strategy for TNBC, particularly for patients unresponsive to conventional treatments.
