Silencing DNMT1 by siRNA enhances the killing effect of NK-92 on Jurkat cells and the underlying mechanism
This study aimed to investigate the effect and mechanism of silencing DNA methyltransferase 1(DNMT1)on the killing effect of NK-92 on Jurkat cells.The DNMT1 expression in the bone marrow of 35 patients with acute lymphocyte leukemia(ALL)from January 2020 to December 2021 were measured.The si-RNA-DNMT1 was transfected into NK-92 cells and co-cultured with Jurkat cells.The killing effect on Jurkat cells was detected by lactate dehydrogenase(LDH)method,and the apoptosis and membrane NKG2D of Jurkat cells were detected by flow cytometry.The levels of IFN-γ were estimated by ELISA.The expressions of Syk,ZAP70,p-Syk,p-ZAP70,perforin and granzyme B in NK-92 cells were measured by Western blotting.The results showed that the level of DNMT1 was increased in patients with ALL compared to the control group(P＜0.05),and it was of diagnostic value for ALL(AUC:0.692,95％CI:0.533-0.851,P=0.033).The progression free survival of patients with high DNMT1 was significantly lower than those with low DNMT1(P＜0.05).NK-92 transfected with si-RNA-DNMT1 showed a higher killing rate and cell apoptosis compared to those of the si-NC group(P＜0.05),along with the enhanced ability to secrete NKG2D and express perforin and granzyme B(P＜0.05).The IFN-γ level of NK-92 transfected with si-RNA-DNMT1 was significantly higher than that in the si-NC group(P＜0.05).The p-Syk and p-ZAP70 protein levels of si-RNA-DNMT1 transfected NK-92 were also significantly higher than those in the si-NC group(P＜0.05).In conclusion,DNMT1 expression increases in ALL and silencing DNMT1 promotes the killing ability of NK-92 cells on Jurkat cells.The mechanism may be related to enhanced function of NK-92 cells and activation of Syk/ZAP70 pathway.
DNA methyltransferase 1acute lymphocyte leukemiaNK-92 cellsJurkat cells
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