# Reversal Effects of 20(R)- and 20(S)-Ginsenoside-Rg3 on Daunorubicin Uptake in Multidrug-Resistant Leukemia Cells Studied in the Single-Cell Biochip

## TL;DR

This study shows how a single-cell biochip can test ginsenosides' ability to reverse drug resistance in leukemia cells, highlighting the role of stereochemistry in drug effectiveness.

## Contribution

The study introduces a microfluidic single-cell biochip to evaluate ginsenoside reversal effects on drug resistance using microgram quantities and single cells.

## Key findings

- Rg3-S is more effective than Rg3-R in increasing daunorubicin accumulation in resistant leukemia cells.
- The SCB platform enables drug resistance studies with minimal compound and cell quantities.
- Stereochemistry of ginsenosides significantly influences their reversal efficacy against P-gp-mediated resistance.

## Abstract

Multidrug resistance (MDR), frequently mediated by overexpression of the P-glycoprotein (P-gp) efflux transporter, remains a major challenge in the treatment of leukemia by limiting intracellular accumulation of chemotherapeutic agents such as daunorubicin (DNR). This study evaluates the applicability of a microfluidic-based single-cell biochip to investigate the reversal effects of microgram-level ginsenosides on daunorubicin uptake in multidrug-resistant leukemia cells. Pure ginsenosides are difficult to obtain in bulk and are typically available only in milligram quantities, which restricts their evaluation using conventional MDR assays such as flow cytometry that require large cell populations and substantial amounts of compounds. To address this limitation, a microfluidic single-cell biochip (SCB) requiring microgram quantities of ginsenosides (<100 µg) and fewer than ten cells was employed. Intracellular DNR accumulation was measured in the CEM/VLB1000 leukemia cell line following treatment with DNR alone or in combination with ginsenoside Rg3-R, ginsenoside Rg3-S, 20(S)-protopanaxatriol (PPT), and 20(S)-protopanaxadiol (PPD), in order to compare their relative efficacy in enhancing drug accumulation. Although Rg3-R and Rg3-S share highly similar chemical structures and are glycosylated derivatives of the PPD aglycone, Rg3-S exhibited greater potency in increasing intracellular daunorubicin accumulation than Rg3-R, and both were more effective than PPD. These findings underscore the importance of ginsenoside stereochemistry modulating P-gp-associated drug resistance and demonstrate the utility of the SCB platform for quantifying daunorubicin accumulation in multidrug-resistant leukemia cells at single-cell resolution.