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Published on: May 19, 2023
Discrimination of Multidrug Resistance in Cancer Cells Achieved Using Single-Cell Analysis
Haiyan Wang1, Runxuan Zhang2, Di Yang2
1Department of Chemistry and Chemical Engineering, Shanxi Datong University, Datong, Shanxi, China. why7135280@126.com.
Abstract:
The biophysical signatures of single cells, such as multidrug resistance (MDR), may easily change during their various disease states. Therefore, there is an ever-growing need for advanced methods to study and analyze the response of cancer cells to therapeutic intervention. To determine the cancer cells and responses to various cancer therapies, from a cell mortality perspective, we report a label-free and real-time method to monitor the in situ responses of ovarian cancer cells using a single-cell bioanalyzer (SCB). The SCB instrument was used to detect different ovarian cancer cells, such as NCI/ADR-RES cells, which are multidrug resistant (MDR), and non-MDR OVCAR-8 cells. The discrimination of ovarian cells has been achieved at the single-cell level by measuring drug accumulation quantitatively in real time, in which the accumulation is high in non-MDR single cells without drug efflux but is low in MDR single cells which are not efflux-free. The SCB was constructed as an inverted microscope for optical imaging and fluorescent measurement of a single cell that was retained in a microfluidic chip. The single ovarian cancer cell retained in the chip offered sufficient fluorescent signals for the SCB to measure the accumulation of daunorubicin (DNR) in the single cell in the absence of cyclosporine A (CsA). The same cell allows us to detect the enhanced drug accumulation due to MDR modulation in the presence of CsA, which is the MDR inhibitor. The measurement of drug accumulation in a cell was achieved after it was captured in the chip for one hour, with the correction of background interference. The detection of accumulation enhancement due to MDR modulation by CsA was determined in terms of either the accumulation rate or enhanced concentration of DNR in the single cell (same cell, p < 0.01). It showed that with the effectiveness of efflux blocking by CsA, the intracellular DNR concentration in a single cell increased by threefold against its same cell control. This single-cell bioanalyzer instrument has the ability to discriminate MDR in different ovarian cells due to drug efflux in them by eliminating the interference of background fluorescence and by using the same cell control.
Insights
Researchers developed a label-free, real-time method using a single-cell bioanalyzer to monitor ovarian cancer cell responses to therapy. This technique effectively distinguishes multidrug-resistant (MDR) from non-MDR cells by measuring drug accumulation, aiding in personalized cancer treatment strategies.
Area of Science:
- Biophysics
- Cell Biology
- Cancer Research
Background:
- Multidrug resistance (MDR) in cancer cells complicates therapeutic interventions.
- Accurate, real-time monitoring of cellular responses to treatment is crucial for effective cancer therapy.
- Existing methods may not adequately capture dynamic changes in cellular drug resistance.
Purpose of the Study:
- To develop and validate a label-free, real-time method for analyzing ovarian cancer cell responses to therapeutic agents.
- To discriminate between multidrug-resistant (MDR) and non-MDR ovarian cancer cells at the single-cell level.
- To quantitatively measure drug accumulation and efflux in individual cancer cells.
Main Methods:
- Utilized a single-cell bioanalyzer (SCB) integrated with a microfluidic chip for in situ monitoring.
- Employed optical imaging and fluorescent measurements to track daunorubicin (DNR) accumulation in single ovarian cancer cells (NCI/ADR-RES and OVCAR-8).
- Assessed MDR modulation by using cyclosporine A (CsA), an MDR inhibitor, and analyzing changes in DNR accumulation within the same cell.
Main Results:
- Successfully discriminated between MDR and non-MDR ovarian cancer cells based on real-time drug accumulation patterns.
- Demonstrated significantly lower DNR accumulation in MDR cells compared to non-MDR cells due to active drug efflux.
- Showed a threefold increase in intracellular DNR concentration in MDR cells when treated with CsA, confirming effective efflux inhibition.
Conclusions:
- The single-cell bioanalyzer (SCB) provides a robust platform for label-free, real-time assessment of single-cell drug responses and MDR.
- This method enables precise quantification of drug accumulation and efflux, facilitating the identification of resistant cancer cell populations.
- The SCB technology holds promise for advancing personalized medicine by guiding therapeutic strategies against ovarian cancer.
