下调SPTBN2通过抑制Nrf2/GPX4信号诱导肺腺癌细胞A549铁死亡

Downregulation of SPTBN2 induced ferroptosis in lung adenocarcinoma cell A549 by inhibiting Nrf2/GPX4 signaling

目的研究下调SPTBN 2对肺腺癌细胞A549铁死亡的影响和机制。方法肺腺癌细胞A549分成Control、si-NC(转染siRNA control)、si-SPTBN 2(转染SPTBN 2 siRNA)、si-Nrf 2(转染Nrf 2 siRNA)、si-SPTBN 2+Vector(转染SPTBN 2 siRNA、阴性对照载体)、si-SPTBN 2+Nrf两组(转染SPTBN 2 siRNA、Nrf 2过表达载体),Western blot检测ACSL4、SLC7A11、Nrf 2、GPX4蛋白表达,CCK-8实验检测增殖,DCFH-DA法检测ROS,比色法检测Fe^(2+)、MDA、4-HNE,微量法检测GSH。结果与Control、si-NC组比较,si-SPTBN 2、si-Nrf两组肺腺癌细胞增殖活性降低,细胞内Fe^(2+)含量升高,细胞中ACSL4、SLC7A11、Nrf 2、GPX4蛋白表达减少,ROS、MDA、4-HNE含量升高,GSH含量降低。与si-SPTBN 2+Vector组比较,si-SPTBN 2+Nrf两组肺腺癌细胞增殖活性升高,细胞Fe^(2+)、ROS、MDA、4-HNE含量降低,GSH含量升高,ACSL4、SLC7A11、Nrf 2、GPX4蛋白表达增多。结论下调SPTBN 2通过抑制Nrf 2/GPX4信号诱导肺腺癌细胞A549铁死亡。

Objective To study the effect and mechanism of down-regulation of SPTBN2 on ferroptosis of lung adenocarcinoma cell A549. Methods A549 lung adenocarcinoma cells were divided into Control, si-NC(transfected with siRNA control), si-SPTBN2(transfected with SPTBN2 siRNA), si-Nrf2(transfected with Nrf 2 siRNA), si-SPTBN2+Vector(transfected with SPTBN2 siRNA, negative control vector), si-SPTBN2+Nrf 2 groups(transfected with SPTBN2 siRNA and Nrf 2 overexpression vector). Western blot was used to detect ACSL4, SLC7A11, Nrf 2, and GPX4 protein expression. CCK-8 experiment was used to detect proliferation. DCFH-DA method wsa used to detect ROS. Colorimetric method was used to detect Fe^(2+), MDA, and 4-HNE. Micro method was used to detect GSH. Results Compared with Control and si-NC groups, lung adenocarcinoma cell proliferation activity in si-SPTBN2 and si-Nrf2 groups was decreased;Fe^(2+)content in cells was increased;ACSL4, SLC7A11, Nrf2, and GPX4 protein expression in cells were decreased;ROS, MDA, and 4-HNE contents were increased;GSH content was decreased. Compared with si-SPTBN2+Vector group, lung adenocarcinoma cell proliferation activity in si-SPTBN2+Nrf 2 group was increased;Fe^(2+), ROS, MDA, and 4-HNE contents were decreased;GSH content was increased;ACSL4, SLC7A11, Nrf 2, and GPX4 protein expression were increased. Conclusion Down-regulation of SPTBN2 induces ferroptosis of lung adenocarcinoma cell A549 by inhibiting Nrf2/GPX4 signal.