[Mechanism of Apoptosis for Corynoline-Mediated Reversing the Drug Resistance of AML HL-60/ADR Cells].

OBJECTIVE

To explore the function and molecular mechanism of corynoline in reversing drug resistance in acute myeloid leukemia cell line HL-60/adriamycin (ADR).

METHODS

HL-60/ADR cells were divided into four groups: control group, ADR group, corynoline group, and ADR+corynoline group. CCK-8 assay was applied to measure the cell viability and drug half maximal inhibitory concentration (IC50), and flow cytometry was used to detect the intensity of intracellular ADR autofluorescence. Transcriptome sequencing was performed to explore its molecular mechanism, and RT-qPCR and Western blot were used to verify the expression of related genes, such as anti-apoptotic gene BCL2 , pro-apoptotic gene BAX and drug-resistant gene MRP1 . The expression of pathway related proteins enriched by differentially expressed genes was detected by Western blot.

RESULTS

Corynoline inhibited the viability of HL-60/ADR cells in a concentration- and time-dependent manner (r =-0.994, -0.981, -0.928). Compared with the ADR group, the ADR autofluorescence in HL-60/ADR cells was significantly enhanced in the ADR+corynoline group. The IC50 in the ADR+corynoline group was 1.150 µmol/L, which was markedly decreased than 8.081 µmol/L in the ADR group. In contrast to the ADR group, the expression of p-AKT was down-regulated and the activity of PI3K/AKT signaling pathway was inhibited in the dual-drug combination group. Moreover, the expression of pro-apoptotic protein BAX was increased, while the expression of anti-apoptotic protein Bcl-2 and drug-resistant protein MRP1 was decreased.

CONCLUSION

Corynoline has the function of reversing the drug resistance of HL-60/ADR cells, which is related to reducing the expression of intracellular drug resistance protein MRP1. Additionally, corynoline inhibits the activity of the PI3K/AKT signaling pathway in HL-60/ADR cells.