[Effect of Sunitinib on Pyroptosis of Drug-Resistant Leukemia K562/ADR Cells and Its Pathway].

To investigate the inducing effect of sunitinib on the death of drug-resistant leukemia K562/ADR cells and the related signaling pathway.K562/ADR cells were treated with different concentrations of sunitinib, and the cells were collected at 24, 48, 72, and 96 hours, respectively. MTS assay was used to detect the effect of sunitinib on the proliferation of K562/ADR cells, and the appropriate sunitinib intervention time and concentration were determined. QPCR and Western blot were used to detect the mRNA and protein expression levels of apoptosis-related genes in K562/ADR cells treated with sunitinib. Four different cell death inhibitors Nec-1, VX-765, CQ and Fer-1 were used to detect the death mode of K562/ADR cells treated with sunitinib. QPCR and Western blot were used to detect the mRNA and protein expression levels of pyroptosis-related genes in K562/ADR cells treated with sunitinib.Sunitinib significantly inhibited the proliferation of K562/ADR cells in a time - and concentration-dependent manner(R48 H=0.9579, r4 μg/ml=0.9740). The IC50 of sunitinib was (3.96±0.14) μg/ml at 48 hours. The mRNA and protein expression levels of apoptosis-related genes Bax, BCL-2 , Caspase-3 and Caspase-9 in K562/ADR cells treated with sunitinib did not change significantly. After treatment with four different cell death inhibitors, only the pyroptosis inhibitor VX-765 could significantly reverse the inhibitory effect of sunitinib on the proliferation of K562/ADR cells (P<0.01). The mRNA and protein expression levels of pyroptosis-related genes Caspase-1, Caspase-4, Caspase-5, NLRP3, GSDMD and IL-1β in K562/ADR cells treated with sunitinib were significantly increased (P<0.01).Sunitinib can induce pyroptosis in drug-resistant leukemia K562/ADR cells. Further study of the signaling pathways related to pyroptosis may provide experimental basis for the treatment of drug-resistant leukemia.舒尼替尼诱导耐药白血病细胞K562/ADR焦亡 的作用及其通路研究.探讨舒尼替尼(SU11248)对耐药白血病细胞K562/ADR死亡的诱导作用及其相关信号通路.使用不同浓度舒尼替尼干预K562/ADR细胞，分别于24、48、72、96 h收集各组细胞，采用MTS法检测舒尼替尼对K562/ADR细胞增殖能力的影响，确定适当的舒尼替尼干预时间和浓度。通过qPCR和Western blot检测舒尼替尼干预后K562/ADR细胞的凋亡相关基因mRNA和蛋白表达水平变化。使用4种不同细胞死亡抑制剂Nec-1、VX-765、CQ、Fer-1检测舒尼替尼干预后K562/ADR细胞的死亡方式。通过qPCR和Western blot检测舒尼替尼干预后K562/ADR细胞的焦亡相关基因mRNA和蛋白表达水平变化.舒尼替尼可明显抑制K562/ADR细胞的增殖，抑制效果呈一定的时间及浓度依赖性(r48 H=0.9579、r4 μg/ml=0.9740)，其48 h的IC50为（3.96±0.14） μg/ml；舒尼替尼干预后K562/ADR细胞凋亡相关基因Bax、BCL-2、Caspase-3、Caspase-9的mRNA和蛋白表达水平均无明显变化；4种不同细胞死亡抑制剂处理后，仅焦亡抑制剂VX-765能够明显逆转舒尼替尼对K562/ADR细胞的增殖抑制作用（P＜0.01）；舒尼替尼干预后K562/ADR细胞焦亡相关基因Caspase-1、Caspase-4、Caspase-5、NLRP3、GSDMD、IL-1β的mRNA和蛋白表达水平均明显升高（P＜0.01）.舒尼替尼可诱导耐药白血病细胞K562/ADR发生焦亡，深入研究细胞焦亡相关信号通路有望为耐药白血病治疗提供实验依据.