Chinese Journal of Experimental Traditional Medical Formulae
Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway
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1.Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China;2.Shanghai Putuo District Central Hospital, Shanghai 200062,China

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    Abstract:

    Objective To investigate the effect and mechanism of saikosaponin A (SSA) on the reversal of cisplatin (DDP) resistance in human lung cancer cell line A549/DDP.Methods The resistance of A549 and A549/DDP cells to DDP and the inhibitory effects of SSA against the proliferation of A549 and A549/DDP cells were detected using cell counting kit-8 (CCK-8). The apoptosis rates of A549/DDP cells treated with SSA or DDP or SSA combined with DDP and the changes in reactive oxygen species (ROS) were determined by flow cytometry. The mRNA expression levels of C-myc, B-cell lymphoma 2 (Bcl-2) and cysteinyl aspartate-specific protease-3 (Caspase-3) were detected by real-time polymerase chain reaction (Real-time PCR), followed by the determination of β-catenin transcriptional activity using the TopFish dual-luciferase reporter assay system and the measurement of β-catenin protein expression in A549/DDP cells by Western blot.Results The results of CCK-8 assay showed that the DDP resistance of A549/DDP cells was 12.82 times that of A549 cells (P<0.05). SSA inhibited the viability of A549 cells with the half maximal inhibitory concentration (IC50) being 34.9 μmol·L-1, and also suppressed the viability of A549/DDP cells in a concentration-dependent manner. Since the inhibition rate of 20 μmol/L SSA against A549/DDP cells was less than 10%, the reversal concentration was set at 20 μmol/L. Flow cytometry revealed that compared with the control, DDP alone increased the apoptosis rate of A549/DDP cells (P<0.05), stimulated the accumulation of intracellular ROS (P<0.05), down-regulated the mRNA expression levels of C-myc and Bcl-2 in A549/DDP cells, up-regulated Caspase-3 mRNA expression, and reduced the transcriptional activity of β-catenin (P<0.05). Compared with the DDP group, the SSA+DDP group exhibited obviously increased apoptosis of A549/DDP cells, enhanced accumulation of intracellular ROS, down-regulated C-myc and Bcl-2 mRNA expression, up-regulated Caspase-3 mRNA expression (P<0.05), and weakened β-catenin transcription (P<0.05). DDP combined with SSA better decreased the β-catenin protein expression in contrast to that of control or DDP (P<0.05).Conclusions SSA enhances the sensitivity of A549/DDP cells to DDP possibly by inhibiting the activation of Wnt/β-catenin pathway.

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History
  • Received:February 06,2021
  • Revised:
  • Adopted:
  • Online: May 06,2021
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