Sirna Interference Nrf2Reversed Resistance Mechanisms Of Imatinib-resistant Cell Lines K562/G01Research

ObjectiveTo investigate the effect and possible mechanism of siRNA-mediated knockdown of the transcription factor nuclear factor E2-related factor2(Nrf2) on drug resistance in imatinib-resistant chronic myeloid leukemia cell line K562/G01cell lines.Methods(1) Nrf2-siRNA mediated by lentivirus was transfected into K562/G01cell lines, and then the transfection efficiency was detected by laser scanning confocal microscope (LSCM) and Flow Cytometry (FCM), the expression of Nrf2mRNA and protein, as well as downstream thioredoxin reductase (TrxR) was detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting respectively.(2) ROS generation was determined by FCM.(3) The drug sensitivity and apoptosis rate was analyzed by Cell Counting Kit-8(CCK-8) assay and FCM.Results(1) In transfected K562/G01cells, the expression of Nrf2and TrxR mRNA and protein was significantly inhibited (P<0.05).(2) In transfected K562/G01cells, the ROS generation was increased(P<0.05).(3) Compared with untreated group, the resistance index was significantly decreased and the apoptosis rate was observably increased in transfected group (P<0.05).Conclusion These results demonstrates that Nrf2protectes K562/G01cells from imatinib-induced oxidative stress and apoptosis, resultes in the drug resistence. Knockdown of Nrf2by siRNA might improve the sensitivity of K562/G01cells to imatinib-induced apoptosis.