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Vimentin-targeting adaptogen withaferin A: Potential to selectively suppress cervical cancer - Single-cell
Ewa Pięta1, Agnieszka Panek1, Monika Szczepanek-Dulska1
1Institute of Nuclear Physics Polish Academy of Sciences, PL-31342, Krakow, Poland.
Abstract:
This study investigates the preferential anticancer effects of withaferin A, an adaptogenic compound, on primary and metastatic cervical cancer cells (C-33 A and CaSki, respectively) and non-cancerous skin fibroblast cells (Detroit-551). Employing a multi-modal approach, we combined biological assays with advanced vibrational spectroscopic imaging techniques, including Fourier-transform infrared (FT-IR), Raman (RS), and atomic force microscopy (AFM). The results revealed a dose-dependent reduction in cell viability, with a more pronounced effect observed in C-33 A cells compared to CaSki and fibroblasts, indicating a heightened sensitivity of C-33 A cells to withaferin A. The comet assay revealed significantly higher levels of DNA damage in primary tumor C-33A cells, whereas minimal DNA breaks were observed in fibroblasts and metastatic cells, further confirming the higher sensitivity of cancer cells compared to fibroblasts. Fluorescence staining and AFM topography imaging showed morphological alterations in cancer cells at higher withaferin A doses and longer incubation times. Flow cytometry analysis revealed significant apoptotic changes in primary C-33A cells due to withaferin A treatment, highlighting a large amount of cells undergoing late apoptosis, compared to a weaker apoptotic effect on metastatic CaSki cells and negligible effect for fibroblasts. Spectroscopic analyses revealed molecular alterations in lipid, protein, and nucleic acid composition, indicative of withaferin A's impact on cellular membranes and genetic material. These findings highlight withaferin A as a promising therapeutic agent with the potential to preferentially target primary cervical cancer cells, while minimizing toxicity to healthy cells.
Insights
Withaferin A preferentially targets primary cervical cancer cells, inducing DNA damage and apoptosis while sparing healthy fibroblasts. This adaptogenic compound shows promise for selective cancer therapy.
Area of Science:
- Oncology
- Biochemistry
- Cell Biology
Background:
- Cervical cancer remains a significant global health concern.
- Developing targeted therapies with minimal side effects is crucial.
- Withaferin A, an adaptogenic compound, has shown potential anticancer properties.
Purpose of the Study:
- To investigate the preferential anticancer effects of withaferin A on primary and metastatic cervical cancer cells.
- To compare its efficacy and toxicity against non-cancerous skin fibroblast cells.
- To elucidate the underlying molecular mechanisms of withaferin A's action.
Main Methods:
- Cell viability assays (MTT).
- Comet assay for DNA damage assessment.
- Fluorescence staining and Atomic Force Microscopy (AFM) for morphological analysis.
- Flow cytometry for apoptosis detection.
- Vibrational spectroscopic imaging (FT-IR, Raman) for molecular composition analysis.
Main Results:
- Withaferin A exhibited a dose-dependent reduction in cell viability, with greater efficacy against primary C-33 A cells than metastatic CaSki cells and fibroblasts.
- Significant DNA damage and apoptosis were observed in primary C-33 A cells, with minimal effects on fibroblasts.
- Spectroscopic analyses revealed molecular alterations in lipids, proteins, and nucleic acids, indicating withaferin A's impact on cellular components.
Conclusions:
- Withaferin A demonstrates preferential anticancer effects on primary cervical cancer cells.
- It induces significant DNA damage and apoptosis in cancer cells while exhibiting lower toxicity to healthy fibroblasts.
- Withaferin A is a promising therapeutic candidate for targeted cervical cancer treatment.
