背景:非小细胞肺癌(NSCLC)占肺癌(LC)病例的80%以上,使其成为全球癌症相关死亡的主要原因。T-box转录因子5(TBX5)是胚胎和器官发育的重要调节因子,在癌症的发生发展中起着关键作用。这里,我们的目的是研究TBX5参与LC细胞内的铁凋亡及其潜在机制。
方法:首先,在人LC细胞中检测TBX5表达。接下来,在A549和NCI-H1703细胞中进行TBX5和Yes1相关转录调节因子(YAP1)的过表达和TEA结构域1(TEAD1)的敲减。A549和NCI-H1703细胞的增殖能力,GSH,MDA,ROS,和Fe2+水平被测量。进行免疫共沉淀(Co-IP)以验证TBX5蛋白是否可以结合YAP1。然后评估TBX5,YAP1,TEAD1,GPX4,p53,FTH1,SLC7A11和PTGS2蛋白水平。最后,我们在体内验证了TBX5对LC细胞铁凋亡的影响。
结果:TBX5在LC细胞中表达下调,特别是在A549和NCI-H1703细胞中。过表达TBX5显著降低A549和NCI-H1703细胞的增殖能力,下调GPX4和GSH水平,并上调MDA,ROS,和Fe2+水平。Co-IP证实TBX5蛋白可以结合YAP1。此外,oe-YAP1促进Lv-TBX5转染的A549和NCI-H1703细胞的增殖能力,上调GPX4和GSH水平,下调MDA,ROS,和Fe2+水平。此外,oe-YAP1在用Lv-TBX5转染的A549和NCI-H1703细胞中促进FTH1和SLC7A11水平并抑制p53和PTGS2水平。然而,用si-TEAD1转染进一步逆转了这些效应。体内实验进一步验证了TBX5促进LC细胞中的铁凋亡。
结论:TBX5抑制YAP1-TEAD1通路的激活,促进LC细胞的铁凋亡。
BACKGROUND: Non-small cell lung cancer (NSCLC) accounts for more than 80 % of lung cancer (LC) cases, making it the primary cause of cancer-related mortality worldwide. T-box transcription factor 5 (TBX5) is an important regulator of embryonic and organ development and plays a key role in cancer development. Here, our objective was to investigate the involvement of TBX5 in ferroptosis within LC cells and the underlying mechanisms.
METHODS: First, TBX5 expression was examined in human LC cells. Next, overexpression of TBX5 and Yes1-associated transcriptional regulator (YAP1) and knockdown of TEA domain 1 (TEAD1) were performed in
A549 and NCI-H1703 cells. The proliferation ability of
A549 and NCI-H1703 cells, GSH, MDA, ROS, and Fe2+ levels were measured. Co-immunoprecipitation (Co-IP) was performed to verify whether TBX5 protein could bind YAP1. Then TBX5, YAP1, TEAD1, GPX4, p53, FTH1, SLC7A11 and PTGS2 protein levels were assessed. Finally, we verified the effect of TBX5 on ferroptosis in LC cells in vivo.
RESULTS: TBX5 expression was down-regulated in LC cells, especially in
A549 and NCI-H1703 cells. Overexpression of TBX5 significantly decreased proliferation ability of A549 and NCI-H1703 cells, downregulated GPX4 and GSH levels, and upregulated MDA, ROS, and Fe2+ levels. Co-IP verified that TBX5 protein could bind YAP1. Moreover, oe-YAP1 promoted proliferation ability of
A549 and NCI-H1703 cells transfected with Lv-TBX5, upregulated GPX4 and GSH levels and downregulated MDA, ROS, and Fe2+ levels. Additionally, oe-YAP1 promoted FTH1 and SLC7A11 levels and inhibited p53 and PTGS2 levels in
A549 and NCI-H1703 cells transfected with Lv-TBX5. However, transfection with si-TEAD1 further reversed these effects. In vivo experiments further validated that TBX5 promoted ferroptosis in LC cells.
CONCLUSIONS: TBX5 inhibited the activation of YAP1-TEAD1 pathway to promote ferroptosis in LC cells.