Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Cell Cycle. 2022 Sep;21(18):1996-2007. doi: 10.1080/15384101.2022.2078615. Epub 2022 May 30.
Cyclophilin A (cypA) is overexpressed in many types of carcinomas, including non-small-cell lung cancer (NSCLC). However, the effect of anoxia, a critical feature of the carcinoma cell microenvironment, on cypA expression in NSCLC is unknown. Here, formaldehyde-fixed and paraffin-embedded samples were collected from 60 subjects with NSCLC. The protein expression levels of cypA and hypoxia-inducible factor-1α (HIF-1α) were evaluated using immunohistochemistry. Kaplan-Meier analysis showed that subjects with high cypA expression had remarkably shorter progression-free survival than those with low cypA expression. Furthermore, cypA expression levels were significantly related to HIF-1α expression levels (Spearman's correlation = 0.34, P < 0.0001). To further assess the effect of cypA, an anoxic carcinoma cell model was established. CypA expression was remarkably upregulated in H1299 and A549 cell lines under hypoxic conditions. Overexpression of cypA restored hypoxia-impaired cell growth and prevented reactive oxygen species (ROS) production and cell death in hypoxic A549 and H1299 cells. However, these phenotypes were not altered by the inactive R55A mutant of cypA. Mechanistic studies demonstrated that cypA can bind to and degrade the tumor suppressor protein TXNIP in H1299 and A549 cells. Restored TXNIP expression in cypA-overexpressed and hypoxic NSCLC cells led to increased ROS levels and apoptotic cell numbers and decreased cell growth compared with cypA-overexpressed and hypoxic NSCLC cells. These findings indicate that anoxia results in an increase in cypA expression in NSCLC. Additionally, cypA served as an oncogene during hypoxia by interacting with TXNIP.
亲环蛋白 A(cypA)在多种癌中过表达,包括非小细胞肺癌(NSCLC)。然而,乏氧(癌细 胞微环境的一个关键特征)对 NSCLC 中 cypA 表达的影响尚不清楚。本研究收集了 60 例 NSCLC 患者的福尔马林固定石蜡包埋样本,采用免疫组织化学法检测 cypA 和缺氧诱导因子-1α(HIF-1α)的蛋白表达水平。Kaplan-Meier 分析显示,cypA 高表达的患者无进展生存期显著短于 cypA 低表达的患者。此外,cypA 表达水平与 HIF-1α 表达水平显著相关(Spearman 相关系数=0.34,P < 0.0001)。为了进一步评估 cypA 的作用,建立了乏氧的癌细 胞模型。在 H1299 和 A549 细胞系中,缺氧条件下 cypA 的表达显著上调。在缺氧的 A549 和 H1299 细胞中,过表达 cypA 恢复了缺氧抑制的细胞生长,并防止了活性氧(ROS)的产生和细胞死亡。然而,cypA 的无活性 R55A 突变体并未改变这些表型。机制研究表明,cypA 可以与肿瘤抑制蛋白 TXNIP 结合并降解之,在 H1299 和 A549 细胞中。在 cypA 过表达和缺氧的 NSCLC 细胞中恢复 TXNIP 的表达,与 cypA 过表达和缺氧的 NSCLC 细胞相比,导致 ROS 水平升高、凋亡细胞数量增加和细胞生长减少。这些发现表明,在 NSCLC 中,乏氧导致 cypA 表达增加。此外,cypA 通过与 TXNIP 相互作用,在缺氧时充当致癌基因。