{Reference Type}: English Abstract
{Title}: [Effect of UVRAG Gene on Ferroptosis Induced by Sorafenib in K562 Cells].
{Author}: Ma YM;Wang Y;Yang M;Cai ZM;Yin XC;
{Journal}: Zhongguo Shi Yan Xue Ye Xue Za Zhi
{Volume}: 32
{Issue}: 3
{Year}: 2024 Jun
暂无{DOI}: 10.19746/j.cnki.issn.1009-2137.2024.03.001
{Abstract}: <B>OBJECTIVE: </B>To explore the effect of UV radiation resistance-associated gene (UVRAG) on ferroptosis induced by sorafenib in leukemia K562 cells.<BR><B>METHODS: </B>K562 cells were treated with 0, 0.625, 1.25, 2.5, 5, 10, and 20 μmol/L sorafenib for 24 or 48 hours, and the cell viability was detected by CCK-8 assay. Flow cytometry technology was used to detect the changes of reactive oxygen species (ROS) in K562 cells treated with 0, 5, and 10 μmol/L sorafenib for 24 hours. Western blot was used to detect the protein expression of GPX4 in K562 cells treated with 0, 5, and 10 μmol/L sorafenib and pretreatment with ferroptosis inhibitor. A recombinant lentiviral vector was used to construct UVRAG overexpression cell line in K562 cells. qPCR and Western blot were used to verify UVRAG gene overexpression, and Western blot detected the effect of UVRAG on the protein expression of GPX4 and HMGB1 after treatment with sorafenib.<BR><B>RESULTS: </B>Different concentrations of sorafenib could significantly inhibit the proliferation of K562 cells, and the cell viability gradually decreased with the increase of concentration (r 24 h=-0.9841, r 48 h=-0.9970). The level of ROS was increased (When the concentration was 10 μmol/L, P <0.001), while the expression of GPX4 protein was decreased in the process of 0, 5, 10 μmol/L sorafenib-induced K562 cell death (P <0.05), and the decrease in GPX4 protein could be partially reversed by pretreatment with ferroptosis inhibitor (P <0.05). Compared with NC group and NC-Sorafenib group, the expression of GPX4 protein was significantly decreased (both P <0.05), while HMGB1 protein was significantly increased (both P <0.05).<BR><B>CONCLUSIONS: </B>Sorafenib can induce ferroptosis in K562 cells, and this process can be promoted by UVRAG.<BR><B>UNASSIGNED: </B>UVRAG基因在索拉非尼诱导的K562细胞铁死亡中的作用.<BR><B>UNASSIGNED: </B>探讨抗紫外线辐射相关基因（UVRAG）在索拉非尼（sorafenib）介导的白血病K562细胞铁死亡中的作用及机制。.<BR><B>UNASSIGNED: </B>用浓度为0、0.625、1.25、2.5、5、10、20 μmol/L的索拉非尼分别处理K562细胞24、48 h后，采用CCK-8试剂检测细胞活力；浓度为0、5、10 μmol/L的索拉非尼处理K562细胞24 h后，流式细胞术检测活性氧的变化情况；Western blot分别检测浓度为0、5、10 μmol/L的索拉非尼干预处理及铁死亡抑制剂预处理K562细胞后GPX4蛋白的表达情况；采用重组慢病毒载体在K562细胞中构建UVRAG过表达稳株，应用qPCR及Western blot验证UVRAG过表达情况及其对索拉非尼干预处理后GPX4、HMGB1蛋白表达情况的影响。.<BR><B>UNASSIGNED: </B>不同浓度的索拉非尼均能显著抑制K562细胞增殖，且随着浓度增高细胞活力逐渐下降（r 24 h=-0.9841，r 48 h=-0.9970）；浓度为0、5、10 μmol/L的索拉非尼诱导K562细胞死亡过程中伴随着活性氧水平的增加（浓度为10 μmol/L时，P <0.001）、GPX4蛋白的下降（P <0.05），铁死亡抑制剂预处理后可以部分逆转索拉非尼诱导的GPX4蛋白下降（P <0.05）；与NC组及NC-sorafenib组相比，OE-sorafenib组GPX4蛋白表达明显下降（均P <0.05），HMGB1蛋白表达明显增加（均P <0.05）。.<BR><B>UNASSIGNED: </B>索拉非尼能诱导K562细胞铁死亡，UVRAG促进了索拉非尼诱导的K562细胞铁死亡过程。.