Zeng Cimei, Huang Denggao, Wang Lei, Liang Haimei, Ma Ximiao
Department of Respiratory and Critical Care Medicine, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, Hainan, China.
Central Laboratory, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, Hainan, China.
Cancer Biol Ther. 2025 Dec;26(1):2501780. doi: 10.1080/15384047.2025.2501780. Epub 2025 May 14.
This study aims to explore the effects of silencing Zic family member 5 (ZIC5) on glucose metabolism and disulfidptosis in lung adenocarcinoma (LUAD) cells.
Data from The Cancer Genome Atlas (TCGA) was used to analyze ZIC5 expression in LUAD and its association with patient outcomes. ZIC5 was silenced in A549 and H1299 cells using siRNA. The expression of ZIC5 mRNA and protein was assessed by qRT-PCR and Western blot. Cell proliferation was evaluated through CCK-8 and 5-ethynyl-2'-deoxyuridine (EdU) assays, while glucose uptake, lactate production, and ATP levels were measured to assess energy metabolism. Seahorse XF analysis was used to evaluate extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Disulfidptosis was assessed through NADP/NADPH ratio, glutathione (GSH) content, GSSG/GSH ratio, and immunofluorescence staining.
ZIC5 is highly expressed in LUAD and is associated with poor patient prognosis. Silencing ZIC5 significantly reduced its mRNA and protein levels in A549 and H1299 cells, markedly inhibited cell proliferation, and led to significant decreases in glucose uptake, lactate production, ATP levels, ECAR, and OCR. Additionally, silencing ZIC5 resulted in an increased NADP/NADPH ratio, decreased GSH levels, and a reduced GSSG/GSH ratio, alongside classic disulfidptosis features.
ZIC5 plays a crucial role in promoting LUAD cell proliferation and energy metabolism while inhibiting disulfidptosis. Silencing ZIC5 markedly suppresses these processes, indicating its potential as a therapeutic target in LUAD.
本研究旨在探讨沉默锌指蛋白家族成员5（ZIC5）对肺腺癌细胞（LUAD）葡萄糖代谢和二硫化物诱导细胞死亡的影响。
利用来自癌症基因组图谱（TCGA）的数据，分析ZIC5在LUAD中的表达及其与患者预后的关系。使用小干扰RNA（siRNA）使A549和H1299细胞中的ZIC5沉默。通过定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法评估ZIC5 mRNA和蛋白质的表达。通过细胞计数试剂盒-8（CCK-8）和5-乙炔基-2'-脱氧尿苷（EdU）检测评估细胞增殖，同时测量葡萄糖摄取、乳酸生成和ATP水平以评估能量代谢。使用海马XF分析评估细胞外酸化率（ECAR）和耗氧率（OCR）。通过烟酰胺腺嘌呤二核苷酸磷酸/还原型烟酰胺腺嘌呤二核苷酸磷酸（NADP/NADPH）比值、谷胱甘肽（GSH）含量、氧化型谷胱甘肽/还原型谷胱甘肽（GSSG/GSH）比值和免疫荧光染色评估二硫化物诱导细胞死亡。
ZIC5在LUAD中高表达，且与患者预后不良相关。沉默ZIC5可显著降低A549和H1299细胞中其mRNA和蛋白质水平，明显抑制细胞增殖，并导致葡萄糖摄取、乳酸生成、ATP水平、ECAR和OCR显著降低。此外，沉默ZIC5导致NADP/NADPH比值升高、GSH水平降低、GSSG/GSH比值降低，同时伴有典型的二硫化物诱导细胞死亡特征。
ZIC5在促进LUAD细胞增殖和能量代谢同时抑制二硫化物诱导细胞死亡中起关键作用。沉默ZIC5可显著抑制这些过程，表明其作为LUAD治疗靶点的潜力。
