Department of Physiology & Pharmacology, Snyder Institute for Chronic Diseases, Lung Health Research Group, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
Department of Medicine, Snyder Institute for Chronic Diseases, Lung Health Research Group, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
PLoS One. 2023 Jun 8;18(6):e0286783. doi: 10.1371/journal.pone.0286783. eCollection 2023.
Roles for the baculoviral inhibitor of apoptosis repeat-containing (BIRC) genes, BIRC2 and BIRC3, may include signaling to the inflammatory transcription factor, nuclear factor-κB (NF-κB) and protection from cell death. However, distinct functions for each BIRC are not well-delineated. Given roles for the epithelium in barrier function and host defence, BIRC2 and BIRC3 expression was characterized in pulmonary epithelial cell lines and primary human bronchial epithelial cells (pHBECs) grown as undifferentiated cells in submersion culture (SC) or as highly differentiated cells at air-liquid interface (ALI). In A549 cells, interleukin-1β (IL1B) and tumor necrosis factor α (TNF) induced BIRC3 mRNA (~20-50-fold), with maximal protein expression from 6-24 h. Similar effects occurred in BEAS-2B and Calu-3 cells, as well as SC and ALI pHBECs. BIRC2 protein was readily detected in unstimulated cells, but was not markedly modulated by IL1B or TNF. Glucocorticoids (dexamethasone, budesonide) modestly increased BIRC3 mRNA and protein, but showed little effect on BIRC2 expression. In A549 cells, BIRC3 mRNA induced by IL1B was unchanged by glucocorticoids and showed supra-additivity with TNF-plus-glucocorticoid. Supra-additivity was also evident for IL1B-plus-budesonide induced-BIRC3 in SC and ALI pHBECs. Using A549 cells, IL1B- and TNF-induced BIRC3 expression, and to a lesser extent, BIRC2, was prevented by NF-κB inhibition. Glucocorticoid-induced BIRC3 expression was prevented by silencing and antagonism of the glucocorticoid receptor. Whereas TNF, but not IL1B, induced degradation of basal BIRC2 and BIRC3 protein, IL1B- and TNF-induced BIRC3 protein remained stable. Differential regulation by cytokines and glucocorticoids shows BIRC2 protein expression to be consistent with roles in rapid signaling events, whereas cytokine-induced BIRC3 may be more important in later effects. While TNF-induced degradation of both BIRCs may restrict their activity, cytokine-enhanced BIRC3 expression could prime for its function. Finally, shielding from glucocorticoid repression, or further enhancement by glucocorticoid, may indicate a key protective role for BIRC3.
杆状病毒凋亡抑制重复包含(BIRC)基因,BIRC2 和 BIRC3 的作用可能包括信号转导到炎症转录因子核因子-κB(NF-κB)和防止细胞死亡。然而,每个 BIRC 的特定功能尚未明确界定。鉴于上皮细胞在屏障功能和宿主防御中的作用,BIRC2 和 BIRC3 的表达在肺上皮细胞系和原代人支气管上皮细胞(pHBEC)中进行了特征描述,这些细胞在浸没培养(SC)中作为未分化细胞或在气液界面(ALI)中作为高度分化细胞生长。在 A549 细胞中,白细胞介素-1β(IL1B)和肿瘤坏死因子-α(TNF)诱导 BIRC3 mRNA(~20-50 倍),蛋白表达的最大时间为 6-24 小时。在 BEAS-2B 和 Calu-3 细胞以及 SC 和 ALI pHBEC 中也发生了类似的效应。BIRC2 蛋白在未刺激的细胞中很容易检测到,但 IL1B 或 TNF 对其表达没有明显调节作用。糖皮质激素(地塞米松、布地奈德)适度增加 BIRC3 mRNA 和蛋白,但对 BIRC2 表达影响不大。在 A549 细胞中,IL1B 诱导的 BIRC3 mRNA 不受糖皮质激素影响,并且与 TNF 加糖皮质激素表现出超加性。在 SC 和 ALI pHBEC 中,IL1B 加布地奈德诱导的 BIRC3 也表现出超加性。使用 A549 细胞,IL1B 和 TNF 诱导的 BIRC3 表达,以及在较小程度上,BIRC2 的表达,通过 NF-κB 抑制而被阻止。BIRC3 的糖皮质激素诱导表达被糖皮质激素受体的沉默和拮抗所阻止。虽然 TNF 而不是 IL1B 诱导基础 BIRC2 和 BIRC3 蛋白的降解,但 IL1B 和 TNF 诱导的 BIRC3 蛋白仍然稳定。细胞因子和糖皮质激素的差异调节表明 BIRC2 蛋白表达与快速信号事件的作用一致,而细胞因子诱导的 BIRC3 可能在后期作用中更为重要。虽然 TNF 诱导的两种 BIRC 的降解可能限制其活性,但细胞因子增强的 BIRC3 表达可能使其功能成熟。最后,免受糖皮质激素抑制或进一步增强糖皮质激素,可能表明 BIRC3 具有关键的保护作用。