«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2025年03期

页码:

362-368

栏目:

论著

出版日期:

2025-06-25

文章信息/Info

Title:

Mechanism of secoisolariciresinol diglucoside influencing apoptosis and ferroptosis in non-small cell lung cancer A549 cells through circRNA HIPK3

作者:

周 玲¹; 肖 颖²; 李秋诗¹; 陈兆毅¹; 李 琪³; 吴园明¹

430080 武汉,武汉市普仁医院呼吸与危重症学科¹
430080 武汉,武汉市普仁医院血液内科²
430080 武汉,武汉科技大学临床第四学院内科教研室³

Author(s):

Zhou Ling¹;  Xiao Ying²;  Li Qiushi¹;  Chen Zhaoyi¹;  Li Qi³;  Wu Yuanming¹.

¹Department of Respiratory and Critical Care Medicine Wuhan Pu Ren Hospital Official Website, Wuhan 430080, China; ²Department of hematology, Wuhan Pu Ren Hospital Official Website, Wuhan 430080, China; ³Department of Internal Medicine, The fourth Clinical College of Wuhan University of Science and Technology, Wuhan 430080, China

关键词:

支气管肺癌;  亚麻木酚素;  环状RNA同源盒相互作用蛋白激酶3;  细胞凋亡;  铁死亡

Keywords:

Bronchogenic carcinoma;  Secoisolariciresinol diglucoside;  Circular RNA homeodomain-interacting protein kinase 3;  Apoptosis;  Ferroptosis

分类号:

R734.2

DOI:

10.3877/cma.j.issn.1674-6902.2025.03.004

摘要:

目的 分析亚麻木酚素(secoisolariciresinol diglucoside, SDG)通过环状RNA同源域相互作用蛋白激酶3(circular RNA homeodomain interacting protein kinase 3, circRNA HIPK3)影响非小细胞肺癌(non-small cell lung cancer, NSCLC)A549细胞凋亡及铁死亡的机制。方法 肺癌A549细胞分为A549组、SDG-L组、SDG-M组、SDG-H组、SDG-H+oe-NC组、SDG-H+oe-circRNA HIPK3组。每组实验重复6次。噻唑蓝实验检测细胞存活率; 荧光定量PCR实验检测circRNA HIPK3、NADPH氧化酶1(nicotinamide adenine dinucleotide phosphate oxidase 1, NOX1)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4, GPX4)、铁蛋白重链1(ferritin heavy chain 1, FTH1)、溶质载体家族7成员11(solute carrier family 7 member 11, SLC7A11)的mRNA表达; 试剂盒检测Fe²⁺、丙二醛(malondialdehyde, MDA)、谷胱甘肽(glutathione, GSH)水平; 2',7'-二氯荧光素二乙醇酯染色法检测活性氧(reactive oxygen species, ROS)水平; 蛋白免疫印记法测定NOX1、GPX4、FTH1、SLC7A11、核因子E2相关因子2(nuclear factor erythroid 2-related factor 2, Nrf2)和血红素氧合酶1(heme oxygenase-1, HO-1)的表达变化; Transwell和原位末端标记实验检测细胞侵袭和凋亡。 结果 不同浓度SDG对人支气管上皮细胞HBE1的存活率无显著影响; 降低A549细胞存活率,SDG-H组细胞存活率(55.29±10.05)%降低最显著,选择40 μmol/L SDG进行后续实验。circRNA HIPK3在A549组(2.36±0.33)细胞中高表达,SDG-H组(1.72±0.26)细胞中低表达。SDG-H组细胞Fe²⁺(4.98±0.93)mmol/L、MDA(5.72±1.26)μmol/g prot、ROS(2.12±0.81)、细胞凋亡率(29.74±10.05)% 高于A549组(3.25±0.49)mmol/L、(3.36±0.72)μmol/g prot、(1.00±0.19)、(13.84±6.17)%(P<0.01),NOX1的mRNA和蛋白表达高于A549组(P<0.01),GSH(3.94±0.63)μmol/g prot、细胞侵袭数(45.92±16.88)个低于A549组(6.08±1.71)μmol/g prot、(117.33±37.14)个(P<0.05),GPX4、FTH1、SLC7A11的mRNA和蛋白表达、Nrf2和HO-1蛋白水平低于A549组(P<0.05)。SDG-H+oe-circRNA HIPK3组细胞Fe²⁺(4.13±0.41)mmol/L、MDA(3.88±0.94)μmol/g prot、ROS(1.49±0.42)、细胞凋亡率(19.16±8.30)%低于SDG-H+oe-NC组(5.11±0.76)mmol/L、(5.65±1.11)μmol/g prot、(2.26±0.67)、(33.21±11.72)%(P<0.05),NOX1的mRNA和蛋白表达低于SDG-H+oe-NC组(P<0.05),GSH(5.87±1.45)μmol/g prot、细胞侵袭数(92.24±22.72)个 高于SDG-H+oe-NC组(4.11±0.70)μmol/g prot、(54.41±19.10)个(P<0.05),GPX4、FTH1、SLC7A11的mRNA和蛋白表达、Nrf2和HO-1蛋白水平高于SDG-H+oe-NC组(P<0.05)。 结论 SDG通过调控circRNA HIPK3,促进NSCLC A549细胞凋亡,抑制细胞侵袭可能与抑制Nrf2/HO-1抗氧化通路激活细胞铁死亡具有意义。

Abstract:

Objective To analyze the mechanism of secoisolariciresinol diglucoside(SDG)affecting apoptosis and ferroptosis of non-small cell lung cancer A549 cells through circular RNA homeodomain interacting protein kinase 3(circRNA HIPK3). Methods Lung cancer A549 cells were divided into A549 group, SDG-L group, SDG-M group, SDG-H group, SDG-H+oe-NC group, SDG-H+oe-circRNA HIPK3 group. Each experiment was repeated 6 times. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to detect the cell viability. The mRNA expressions of circRNA HIPK3, nicotinamide adenine dinucleotide phosphate oxidase 1(NOX1), glutathione peroxidase 4(GPX4), ferritin heavy chain 1(FTH1), and solute carrier family 7 member 11(SLC7A11). Kits were used to detect the levels of Fe²⁺, malondialdehyde(MDA), and glutathione(GSH). The 2',7'-dichlorofluorescin diacetate staining method was used to detect the level of reactive oxygen species(ROS). Western blot was used to determine the expression changes of NOX1, GPX4, FTH1, SLC7A11, nuclear factor erythroid 2-related factor 2(Nrf2), and heme oxygenase-1(HO-1). The Transwell assay and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were used to detect cell invasion and apoptosis. Results SDG at different concentrations had no significant effect on the survival rate of human bronchial epithelial cells HBE1, but it decreased the survival rate of A549 cells. The cell viability in the SDG-H group [(55.29±10.05)%] decreased most significantly, and 40 μmol/L of SDG was selected for subsequent experiments. Circular RNA HIPK3 was highly expressed in the cells of the A549 group(2.36±0.33)and lowly expressed in the cells of the SDG-H group(1.72±0.26). The levels of Fe²⁺, MDA, ROS and the cell apoptosis rate in the SDG-H group were higher than those in the A549 group(P<0.01). The mRNA and protein expressions of NOX1 were higher than those in the A549 group(P<0.01). The levels of GSH and the number of cell invasions(P<0.05). The mRNA and protein expressions of GPX4, FTH1, SLC7A11, and the protein levels of Nrf2 and HO-1 were lower than those in the A549 group(P<0.05). The levels of Fe²⁺, MDA, ROS and the cell apoptosis rate in the cells of the SDG-H+oe-circRNA HIPK3 group were lower than those in the SDG-H+oe-NC group(P<0.05). The mRNA and protein expressions of NOX1 were lower than those in the SDG-H+oe-NC group(P<0.05). The levels of GSH and the number of cell invasions were higher than those in the SDG-H+oe-NC group(P<0.05). The mRNA and protein expressions of GPX4, FTH1, SLC7A11, and the protein levels of Nrf2 and HO-1 were higher than those in the SDG-H+oe-NC group(P<0.05). Conclusion SDG promoted apoptosis of non-small cell lung cancer A549 cells and inhibited cell invasion by regulating circRNA HIPK3. This might be related to inhibiting the activation of the Nrf2/HO-1 antioxidant pathway and activating ferroptosis in cells.

参考文献/References:

1 Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: Globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249.
2 管懋莹, 周 蕾, 徐蔚杰, 等. 中医及中西医结合治疗晚期非小细胞肺癌的回顾性队列研究[J]. 世界中医药, 2024, 19(17): 2641-2646.
3 余 华, 丁桂清, 马金昀, 等. 中药调控肿瘤干细胞作用机制研究进展[J]. 中成药, 2024, 46(9): 3013-3019.
4 颜雪茹, 何恩鹏, 蔡啸镝. 亚麻木酚素在干预疾病中涉及的信号通路及作用机制[J]. 中国生物化学与分子生物学报, 2024, 40(8): 1057-1064.
5 Dobrowolska K, Regulska-Ilow B. The legitimacy of using dietary supplement diglycoside secoisolariciresinol(SDG)from flaxseed in cancer[J]. Rocz Panstw Zakl Hig, 2021, 72(1): 9-20.
6 王 芹, 何梦婕, 龙方懿. 铁死亡相关的环状RNA在疾病中的作用及机制[J]. 中国生物化学与分子生物学报, 2024, 40(3): 295-311.
7 Xie Y, Yuan X, Zhou W, et al. The circular RNA HIPK3(circHIPK3) and its regulation in cancer progression: Review[J]. Life Sci, 2020, 254: 117252.
8 Bade BC, Dela CC. Lung Cancer 2020: Epidemiology, etiology, and prevention[J]. Clin Chest Med, 2020, 41(1): 1-24.
9 刘亨娅, 刘廷挺, 昌建波. 肺癌根治术后患者生活质量纵向变化轨迹及影响因素[J/CD]. 中华肺部疾病杂志(电子版), 2022, 15(2): 230-233.
10 Scherbakov AM, Stasevich OV, Salnikova DI, et al. Antiestrogenic and antiproliferative potency of secoisolariciresinol diglucoside derivatives on MCF-7 breast cancer cells[J]. Nat Prod Res, 2021, 35(24): 6099-6105.
11 Tannous S, Haykal T, Dhaini J, et al. The anti-cancer effect of flaxseed lignan derivatives on different acute myeloid leukemia cancer cells[J]. Biomed Pharmacother, 2020, 132: 110884.
12 Chen T, Wang Z, Zhong J, et al. Secoisolariciresinol diglucoside induces pyroptosis by activating caspase-1 to cleave GSDMD in colorectal cancer cells[J]. Drug Dev Res, 2022, 83(5): 1152-1166.
13 许 胜, 周露玙, 王 昆. 环状RNA及其作为疾病标志物的潜能[J]. 中国生物化学与分子生物学报, 2018, 34(2): 117-128.
14 Tang Y, Liu J, Li X, et al. Exosomal circrna hipk3 knockdown inhibited cell proliferation and metastasis in prostate cancer by regulating mir-212/bmi-1 pathway[J]. J Biosci, 2021, 46: 69.
15 Si X, Zheng H, Wei G, et al. Circrna hipk3 induces cardiac regeneration after myocardial infarction in mice by binding to notch1 and mir-133a[J]. Mol Ther Nucleic Acids, 2020, 21: 636-655.
16 王宇恒, 陈 亮. 环状RNA Circ HIPK3在非小细胞肺癌中的研究进展[J]. 中国肺癌杂志, 2024, 27(8): 629-636.
17 Zhu Y, Shen L, Xia Q, et al. Extracellular vesicle-derived circHIPK3:Novel diagnostic biomarker for lung cancer[J]. Adv Med Sci, 2023, 68(2): 426-432.
18 魏 强, 冯 昊, 曹伯雄, 等. circSEPT9通过降低miR-345-5p水平调控肺癌A549细胞增殖、迁移及侵袭的分子机制[J]. 中国老年学杂志, 2024, 44(16): 4065-4068.
19 何 苗, 姚文秀, 谷涉群. 溪黄草对circ_0091579/miR-515-5p、肺癌细胞A549增殖及凋亡的影响[J]. 中南医学科学杂志, 2023, 51(5): 660-664.
20 赵世鸿, 陈 键, 高嘉营, 等. 铁死亡在海水诱导支气管上皮细胞损伤中的作用研究[J/CD]. 中华肺部疾病杂志(电子版), 2023, 16(6): 756-760.
21 杨翘伊, 张春云, 孙 硕, 等. 土贝母苷乙通过诱导铁自噬抑制非小细胞肺癌细胞增殖[J]. 中国病理生理杂志, 2024, 40(10): 1834-1843.
22 倪银芸, 杨瑛, 张 立. 抑制肺鳞癌靶点HMGCS1促进细胞铁死亡[J]. 中国肺癌杂志, 2024, 27(5): 330-336.
23 江小红, 库 雄, 魏从兵. 基于Nrf2/GPX4介导的铁死亡探讨异荭草素对非小细胞肺癌细胞增殖和凋亡的影响[J]. 湖南中医药大学学报, 2025, 45(3): 445-452.
24 李温庭, 史志周. 环状RNA调控肿瘤细胞铁死亡的研究进展[J]. 中国病理生理杂志, 2022, 38(8): 1533-1536.
25 李江涛, 李 争, 李翔鹏, 等. 基于Nrf2/GPX4铁死亡途径探讨甘露清瘟方对急性肺损伤大鼠的保护作用[J]. 中成药, 2025, 47(2): 596-600.
26 蔡华俊, 陈致岐, 胡文婷, 等. 鼠曲草总黄酮通过激活Nrf2/SLC7A11/GPX-4信号通路抑制肝细胞铁死亡缓解对乙酰氨基酚诱导的小鼠急性肝损伤[J]. 南方医科大学学报, 2024, 44(11): 2201-2208.
27 常 琳, 王 莹, 程文政, 等. CircRNA通过铁死亡参与调控疾病发生发展的研究进展[J]. 生命科学, 2023, 35(8): 1023-1033.
28 范丽霞, 廖小红, 黄育生, 等. 仙鹤草提取物调控铁死亡抑制肺癌A549细胞增殖的作用及机制研究[J/CD]. 中华中医药学刊, 2024: 1-13
29 袁育珺, 曹华华, 赵 敏, 等. 基于Nrf2-HO-1/GPX4信号轴探讨中药靛玉红衍生物E804抑制肺癌A549细胞增殖和迁移的作用机制[J]. 安徽医科大学学报, 2024, 59(2): 331-335.
30 武小杰, 杨 硕, 查 干, 等. Danthron通过靶向GRIM-12影响Nrf2/HO-1通路缓解COPD中的氧化应激和炎症反应[J]. 华中科技大学学报(医学版), 2025, 54(1): 1-7.
31 赵其辉, 周湘华, 唐东华, 等. 二氢杨梅素通过Nrf2/HO-1信号通路抑制地塞米松诱发的成骨细胞系MC3T3-E1细胞铁死亡[J]. 中国现代应用药学, 2024, 41(23): 3347-3354.
32 王依蕾, 洪 婷, 曾海荣, 等. 麦冬皂苷B促进细胞铁死亡抑制非小细胞肺癌A549细胞增殖的机制[J]. 中国医院药学杂志, 2022, 42(19): 1983-1989.
33 苏 帆, 刘 单, 邓述恺. 去甲斑蝥素通过Nrf2/HO-1通路介导铁死亡抑制小细胞肺癌H446细胞增殖[J]. 现代肿瘤医学, 2025, 33(4): 583-589.

备注/Memo

备注/Memo:

基金项目: 中国金属学会治金安全与健康分会健康卫生科研项目(jkws202411)
通信作者: 肖 颖, Email: 244646755@qq.com

常用功能

更新日期/Last Update: 2025-06-25