鸢尾素调控EBF3/ALOX15通路影响肺腺癌细胞增殖和迁移

doi:10.11958/20242182

天津医药 ›› 2025, Vol. 53 ›› Issue (4): 337-342.doi: 10.11958/20242182

• 细胞与分子生物学 • 下一篇

鸢尾素调控EBF3/ALOX15通路影响肺腺癌细胞增殖和迁移

苏红见¹(), 张春艳², 张卫东³, 韩利¹, 乔亚红¹^,^△()

1 河南省胸科医院呼吸与危重症二病区（邮编450000）
2 河南省胸科医院呼吸与危重症三病区（邮编450000）
3 河南省胸科医院胸外七病区（邮编450000）

收稿日期:2024-12-10 修回日期:2025-01-15 出版日期:2025-04-15 发布日期:2025-04-17
通讯作者: ^△ E-mail:qyh13598037502@163.com
作者简介:苏红见（1984），女，副主任医师，主要从事肺癌及慢性阻塞性肺疾病方面研究。E-mail：suhjmail@163.com
基金资助:
河南省科技攻关项目(242102311079)

Irisin affects the proliferation and migration of lung adenocarcinoma cells by regulating the EBF3/ALOX15 pathway

SU Hongjian¹(), ZHANG Chunyan², ZHANG Weidong³, HAN Li¹, QIAO Yahong¹^,^△()

1 The Second Ward of Respiratory and Critical Care Department, Henan Provincial Chest Hospital, Zhengzhou 450000, China
2 the Third Ward of Respiratory and Critical Care Department, Henan Provincial Chest Hospital, Zhengzhou 450000, China
3 the Seventh Ward of Thoracic Surgery, Henan Provincial Chest Hospital, Zhengzhou 450000, China

Received:2024-12-10 Revised:2025-01-15 Published:2025-04-15 Online:2025-04-17
Contact: ^△ E-mail:qyh13598037502@163.com

苏红见, 张春艳, 张卫东, 韩利, 乔亚红. 鸢尾素调控EBF3/ALOX15通路影响肺腺癌细胞增殖和迁移[J]. 天津医药, 2025, 53(4): 337-342.

SU Hongjian, ZHANG Chunyan, ZHANG Weidong, HAN Li, QIAO Yahong. Irisin affects the proliferation and migration of lung adenocarcinoma cells by regulating the EBF3/ALOX15 pathway[J]. Tianjin Medical Journal, 2025, 53(4): 337-342.

摘要/Abstract

摘要：

目的探讨鸢尾素对早期B细胞因子3（EBF3）/花生四烯酸-15-脂加氧酶（ALOX15）通路的调控作用及肺腺癌细胞增殖和迁移的影响。方法将A549细胞分为鸢尾素溶剂组、鸢尾素组、sh-NC组、EBF3抑制剂（sh-EBF3）组、鸢尾素+sh-NC组、鸢尾素+sh-EBF3组。5-溴-2-脱氧尿嘧啶（EdU）染色、CCK-8法检测细胞增殖；划痕实验检测细胞划痕愈合率；2′，7′-二氯荧光二乙酸酯（DCFH-DA）染色检测细胞中活性氧（ROS）水平；比色法检测细胞中谷胱甘肽（GSH）、丙二醛（MDA）、细胞亚铁（Fe²⁺）水平；透射电镜观察A549细胞中线粒体形态；qRT-PCR检测A549细胞中增殖性细胞核抗原（PCNA）、基质金属蛋白酶2（MMP-2）、谷胱甘肽过氧化酶4（GPX4）mRNA水平；Western blot检测细胞中EBF3、ALOX15蛋白。结果与鸢尾素溶剂组比较，鸢尾素组A549细胞线粒体呈现出铁死亡特征，EdU阳性率、OD₄₅₀值、划痕愈合率、GSH水平、PCNA、MMP-2、GPX4 mRNA水平降低，ROS相对荧光强度、MDA、Fe²⁺水平及EBF3、ALOX15蛋白升高（P<0.05）；与sh-NC组比较，sh-EBF3组A549细胞线粒体呈现出铁死亡现象有所减弱，EdU阳性率、OD₄₅₀值、划痕愈合率、GSH水平、PCNA、MMP-2、GPX4 mRNA水平升高，ROS相对荧光强度、MDA、Fe²⁺水平及EBF3、ALOX15蛋白降低（P<0.05）；sh-EBF3逆转了鸢尾素对A549细胞铁死亡、增殖与迁移的影响。结论鸢尾素可能通过激活EBF3/ALOX15通路诱导A549细胞铁死亡，抑制细胞增殖与迁移。

关键词: 肺腺癌, 细胞增殖, 细胞运动, 铁死亡, 鸢尾素, 早期B细胞因子3/花生四烯酸-15-脂加氧酶通路

Abstract:

Objective To investigate the effect of irisin regulating early B cytokine 3 (EBF3)/arachidonic acid-15-lipoxygenase (ALOX15) pathway on the proliferation and migration of lung adenocarcinoma cells. Methods A549 cells were assigned into the irisin solvent group, the irisin group, the sh-NC group, the EBF3 inhibitor (sh-EBF3) group, the irisin+sh-NC group and the irisin+sh-EBF3 group randomly. 5-bromo-2-deoxyuracil (EdU) staining and CCK-8 method were applied to detect cell proliferation. 5-Scratch experiment was applied to detect the scratch healing rate. 2', 7'-dichlorofluorescein diacetate (DCFH-DA) staining was applied to detect the level of reactive oxygen species (ROS) in cells. The reagent kit was used to detect glutathione (GSH), malondialdehyde (MDA) and ferrous ion (Fe²⁺) in cells. Transmission electron microscopy was applied to observe mitochondrial morphology in A549 cells. QRT-PCR was applied to detect mRNA levels of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase 2 (MMP-2) and glutathione peroxidase 4 (GPX4) in A549 cells. Western blot assay was applied to detect EBF3 and ALOX15 proteins in cells. Results Compared with the irisin solvent group, the mitochondria of A549 cells in the irisin group showed ferroptosis characteristics, the positive rate of EdU, OD₄₅₀ value, scratch healing rate, GSH level, PCNA, MMP-2 and GPX4 mRNA levels decreased, and the ROS relative fluorescence intensity, MDA, Fe²⁺ level, and EBF3 and ALOX15 protein levels increased (P<0.05). Compared with the sh-NC group, the mitochondrial ferroptosis phenomenon of A549 cells was reduced in the sh-EBF3 group, the positive rate of EdU, OD₄₅₀ value, scratch healing rate, GSH level, PCNA, MMP-2 and GPX4 mRNA levels increased, and the ROS relative fluorescence intensity, MDA, Fe²⁺ levels and EBF3 and ALOX15 protein levels reduced (P<0.05). Sh-EBF3 reversed the effect of irisin on ferroptosis, proliferation and migration of A549 cells. Conclusion Irisin may induce ferroptosis in A549 cells and inhibit cell proliferation and migration by activating the EBF3/ALOX15 pathway.

Key words: adenocarcinoma of lung, cell proliferation, cell movement, ferroptosis, irisin, early B-cell factor 3/arachidonate-15-lipoxygenase pathway

中图分类号:

R734.2

图/表 9

参考文献 23

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组别	EdU阳性率/%	OD₄₅₀值	划痕愈合率/%
鸢尾素溶剂组	28.96±1.43	0.83±0.07	35.62±1.83
鸢尾素组	13.25±0.67^a	0.32±0.03^a	16.77±0.79^a
sh-NC组	29.05±1.39	0.83±0.08	35.16±1.72
sh-EBF3组	36.65±1.78^b	0.99±0.06^b	43.39±2.01^b
鸢尾素+sh-NC组	12.99±0.61	0.30±0.02	16.67±0.81
鸢尾素+sh-EBF3组	20.58±0.97^c	0.67±0.05^c	25.59±1.23^c
F	369.847^＊＊	158.605^＊＊	329.832^＊＊

组别	EdU阳性率/%	OD₄₅₀值	划痕愈合率/%
鸢尾素溶剂组	28.96±1.43	0.83±0.07	35.62±1.83
鸢尾素组	13.25±0.67^a	0.32±0.03^a	16.77±0.79^a
sh-NC组	29.05±1.39	0.83±0.08	35.16±1.72
sh-EBF3组	36.65±1.78^b	0.99±0.06^b	43.39±2.01^b
鸢尾素+sh-NC组	12.99±0.61	0.30±0.02	16.67±0.81
鸢尾素+sh-EBF3组	20.58±0.97^c	0.67±0.05^c	25.59±1.23^c
F	369.847^＊＊	158.605^＊＊	329.832^＊＊

组别	ROS/相对荧光强度	MDA/ （nmol/mg）	GSH/ （nmol/mg）	Fe²⁺/ （nmol/mg）
鸢尾素溶剂组	1.00±0.01	6.25±0.32	1.86±0.14	0.47±0.05
鸢尾素组	3.65±0.17^a	17.73±0.85^a	0.65±0.05^a	2.16±0.12^a
sh-NC组	0.98±0.02	6.25±0.92	1.85±0.08	0.43±0.03
sh-EBF3组	0.39±0.03^b	4.16±0.21^b	2.39±0.11^b	0.21±0.01^b
鸢尾素+sh-NC组	3.71±0.18	17.82±0.83	0.62±0.06	2.20±0.13
鸢尾素+sh-EBF3组	1.62±0.07^c	10.51±0.52^c	1.34±0.51^c	1.46±0.12^c
F	1 102.695^＊＊	490.363^＊＊	60.399^＊＊	597.505^＊＊

组别	ROS/相对荧光强度	MDA/ （nmol/mg）	GSH/ （nmol/mg）	Fe²⁺/ （nmol/mg）
鸢尾素溶剂组	1.00±0.01	6.25±0.32	1.86±0.14	0.47±0.05
鸢尾素组	3.65±0.17^a	17.73±0.85^a	0.65±0.05^a	2.16±0.12^a
sh-NC组	0.98±0.02	6.25±0.92	1.85±0.08	0.43±0.03
sh-EBF3组	0.39±0.03^b	4.16±0.21^b	2.39±0.11^b	0.21±0.01^b
鸢尾素+sh-NC组	3.71±0.18	17.82±0.83	0.62±0.06	2.20±0.13
鸢尾素+sh-EBF3组	1.62±0.07^c	10.51±0.52^c	1.34±0.51^c	1.46±0.12^c
F	1 102.695^＊＊	490.363^＊＊	60.399^＊＊	597.505^＊＊

组别	PCNA mRNA	MMP-2 mRNA	GPX4 mRNA
鸢尾素溶剂组	1.00±0.00	1.00±0.00	1.00±0.00
鸢尾素组	0.46±0.03^a	0.36±0.03^a	0.23±0.02^a
sh-NC组	0.99±0.01	1.02±0.02	1.01±0.01
sh-EBF3组	1.58±0.12^b	1.93±0.18^b	2.16±0.17^b
鸢尾素+sh-NC组	0.44±0.03	0.38±0.03	0.25±0.02
鸢尾素+sh-EBF3组	0.87±0.07^c	0.65±0.05^c	0.79±0.06^c
F	300.906^＊＊	331.705^＊＊	537.399^＊＊

鸢尾素调控EBF3/ALOX15通路影响肺腺癌细胞增殖和迁移

Irisin affects the proliferation and migration of lung adenocarcinoma cells by regulating the EBF3/ALOX15 pathway

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组别	EBF3/GAPDH	ALOX15/GAPDH
鸢尾素溶剂组	0.65±0.07	0.33±0.04
鸢尾素组	1.82±0.14^a	0.95±0.07^a
sh-NC组	0.65±0.05	0.29±0.02
sh-EBF3组	0.31±0.03^b	0.11±0.01^b
鸢尾素+sh-NC组	1.79±0.12	0.92±0.08
鸢尾素+sh-EBF3组	1.01±0.09^c	0.50±0.04^c
F	286.983^＊＊	289.120^＊＊

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