GPX1基因在肺癌中的表达特征及其对肺腺癌细胞增殖、迁移、侵袭、凋亡的影响

doi:10.6040/j.issn.1671-7554.0.2025.1424

摘要/Abstract

摘要： 目的探讨谷胱甘肽过氧化物酶1(glutathione peroxidase-1, GPX1)在不同病理类型肺癌细胞系中的表达特征、亚细胞定位以及对肺腺癌细胞生物学功能的影响。方法分别采用实时荧光定量PCR和蛋白质免疫印迹检测正常肺上皮细胞HPAEpic、肺腺癌细胞系A549和H292、肺鳞癌细胞系Calu1、大细胞肺癌细胞系95D、小细胞肺癌细胞系H446中GPX1在mRNA和蛋白水平的表达情况,通过方差分析对比不同病理类型肺癌细胞系与正常肺上皮细胞之间的GPX1表达差异;通过免疫荧光染色明确GPX1在肺腺癌细胞中的亚细胞定位;构建肺腺癌细胞系H292和A549 GPX1基因稳定敲低细胞株,于mRNA和蛋白水平验证敲低效率。分别采用CCK-8实验、划痕实验、Transwell侵袭实验和流式细胞术检测H292和A549细胞株增殖、迁移、侵袭能力和细胞凋亡。结果实时荧光定量PCR结果显示,与正常肺上皮细胞HPAEpic相比,GPX1基因在肺腺癌细胞系H292中的表达上调(P=0.006)。蛋白质免疫印迹结果显示,与正常肺上皮细胞HPAEpic相比,GPX1在多种肺癌细胞系(A549、H292、Calu1、95D、H446)中表达均上调(P<0.001)。CCK-8实验、划痕实验、Transwell侵袭实验、流式细胞术结果显示,敲低GPX1基因表达后,H292和A549细胞增殖(P<0.001)、迁移(P<0.001,P=0.002)、侵袭能力均减弱(P=0.039,P=0.014),细胞凋亡增加(P=0.008,P=0.040)。结论 GPX1基因在多种病理类型肺癌细胞系中高表达,且在肺腺癌细胞系中表达水平最高,GPX1主要定位于细胞质。敲低GPX1基因表达可显著抑制A549和H292肺腺癌细胞株的增殖、迁移及侵袭能力,并促进细胞凋亡。

关键词: 肺腺癌, 谷胱甘肽过氧化物酶1, 增殖, 迁移, 侵袭, 凋亡

Abstract: Objective To investigate the expression characteristics, subcellular localization of glutathione peroxidase-1(GPX1)in different pathological types of lung cancer cell lines and the influence on the biological functions of lung adenocarcinoma cell lines. Methods The expressions of GPX1 at mRNA and protein levels in normal lung epithelial cells HPAEpic, lung adenocarcinoma cell line A549 and H292, lung squamous cell carcinoma cell line Calu1, large cell lung cancer cell line 95D, and small cell lung cancer cell line H446 were detected by quantitative real-time PCR and Western blotting, respectively. The expression differences of GPX1 between different pathological types of lung cancer cell lines and normal lung epithelial cells were compared through analysis of variance. The subcellular localization of GPX1 in lung adenocarcinoma cell lines was clarified by immunofluorescence staining. The lung adenocarcinoma cell lines H292 and A549 GPX1 stably knocked down cell strains were constructed, and the knockdown efficiency was verified at mRNA and protein levels, respectively. The proliferation, migration, invasion abilities and apoptosis of H292 and A549 cell strains were detected by CCK-8 assay, wound healing assay, Transwell assay and flow cytometry, respectively. Results The quantitative real-time PCR results showed that the expression of GPX1 was upregulated in lung adenocarcinoma cell line H292 compared with the normal lung epithelial cells HPAEpic(P=0.006). The results of Western blotting showed that compared with the normal lung epithelial cells HPAEpic, expression of GPX1 was upregulated in multiple lung cancer cell lines(A549, H292, Calu1, 95D, and H446)(P<0.001). The results of CCK-8, wound healing assay, Transwell assay and flow cytometry showed that after knockdown of GPX1, the proliferation(P<0.001), migration(P<0.001, P=0.002)and invasion abilities(P=0.039, P=0.014)of H292 and A549 cell strains were all weakened, while apoptosis rate increased(P=0.008, P=0.040). Conclusion The expression of GPX1 is highly upregulated in various pathological types of lung cancer cell lines, and its expression level is the highest among lung adenocarcinoma cell lines. GPX1 is mainly located in cytoplasm. Down-regulation of GPX1 expression can significantly inhibit the proliferation, migration and invasion abilities of A549 and H292 lung adenocarcinoma cell strains, and promote cell apoptosis.

Key words: Lung adenocarcinoma, Glutathione peroxidase-1, Proliferation, Migration, Invasion, Apoptosis

中图分类号:

R574

殷悦,莫振飞,吴培昕,刘金霞,魏元辉,任佳博,李春笋. GPX1基因在肺癌中的表达特征及其对肺腺癌细胞增殖、迁移、侵袭、凋亡的影响[J]. 山东大学学报 (医学版), 2026, 64(1): 65-73.

YIN Yue, MO Zhenfei, WU Peixin, LIU Jinxia, WEI Yuanhui, REN Jiabo, LI Chunsun. Expression of GPX1 in lung cancer cell lines and its influence on the proliferation, migration, invasion abilities and apoptosis in lung adenocarcinoma cell lines[J]. Journal of Shandong University (Health Sciences), 2026, 64(1): 65-73.

参考文献

[1] Han G, Sinjab A, Rahal Z, et al. An atlas of epithelial cell states and plasticity in lung adenocarcinoma [J]. Nature, 2024, 627(8004): 656-663.
[2] Succony L, Rassl DM, Barker AP, et al. Adenocarcinoma spectrum lesions of the lung: detection, pathology and treatment strategies [J]. Cancer Treat Rev, 2021, 99: 102237. doi: 10.1016/j.ctrv.2021.102237
[3] Jeon H, Wang S, Song J, et al. Update 2025: management of nonsmall-cell lung cancer [J]. Lung, 2025, 203(1): 53. doi: 10.1007/s00408-025-00801-x
[4] Zhou F, Guo H, Xia Y, et al. The changing treatment landscape of EGFR-mutant nonsmall-cell lung cancer [J]. Nat Rev Clin Oncol, 2025, 22(2): 95-116.
[5] Jelic MD, Mandic AD, Maricic SM, et al. Oxidative stress and its role in cancer [J]. J Cancer Res Ther, 2021, 17(1): 22-28.
[6] Humphries BA, Zhang A, Buschhaus JM, et al. Enhanced mitochondrial fission inhibits triple-negative breast cancer cell migration through an ROS-dependent mechanism [J]. iScience, 2023, 26(6): 106788. doi: 10.1016/j.isci.2023.106788
[7] Cheung EC, Vousden KH. The role of ROS in tumour development and progression [J]. Nat Rev Cancer, 2022, 22(5): 280-297.
[8] Zhao Y, Wang H, Zhou J, et al. Glutathione peroxidase GPX1 and its dichotomous roles in cancer[J]. Cancers(Basel), 2022, 14(10): 2560.doi: 10.3390/cancers14102560
[9] Meng Q, Shi S, Liang C, et al. Abrogation of glutathione peroxidase-1 drives EMT and chemoresistance in pancreatic cancer by activating ROS-mediated Akt/GSK3 β/Snail signaling [J]. Oncogene, 2018, 37(44): 5843-5857.
[10] 包舒晴, 杨明月, 刘端瑞, 等. NOX4在幽门螺旋杆菌诱导胃癌细胞ROS中的作用 [J]. 山东大学学报(医学版), 2022, 60(6): 19-25. BAO Shuqing, YANG Mingyue, LIU Duanrui, et al. Role of NOX4 in ROS induction by Helicobacter pylori in gastric cancer cells[J]. Journal of Shandong University(Health Sciences), 2022, 60(6): 19-25.
[11] Huang C, Ding G, Gu C, et al. Decreased selenium-binding protein 1 enhances glutathione peroxidase 1 activity and downregulates HIF-1α to promote hepatocellular carcinoma invasiveness [J]. Clin Cancer Res, 2012, 18(11): 3042-3053.
[12] Huang Y, Fang W, Wang Y, et al. Transforming growth factor-β1 induces glutathione peroxidase-1 and protects from H₂O₂-induced cell death in colon cancer cells via the Smad2/ERK1/2/HIF-1α pathway [J]. Int J Mol Med, 2012, 29(5): 906-912.
[13] Gan X, Chen B, Shen Z, et al. High GPX1 expression promotes esophageal squamous cell carcinoma invasion, migration, proliferation and cisplatin-resistance but can be reduced by vitamin D [J]. Int J Clin Exp Med, 2014, 7(9): 2530-2540.
[14] Handy DE, Loscalzo J. The role of glutathione peroxidase-1 in health and disease [J]. Free Radic Biol Med, 2022, 188: 146-61. doi: 10.1016/j.freeradbiomed.2022.06.004
[15] He R, Jiang H, Zhang C, et al. CXCL16 promotes proliferation of head and neck squamous cell carcinoma by regulating GPX1-mediated antioxidant levels [J]. J Zhejiang Univ Sci B, 2024, 26(1): 92-106.
[16] Lee E, Choi A, Jun Y, et al. Glutathione peroxidase-1 regulates adhesion and metastasis of triple-negative breast cancer cells via FAK signaling[J]. Redox Biol, 2020, 29: 101391. doi: 10.1016/j.redox.2019.101391
[17] Gong TT, Guo Q, Li X, et al. Isothiocyanate iberin inhibits cell proliferation and induces cell apoptosis in the progression of ovarian cancer by mediating ROS accumulation and GPX1 expression [J]. Biomed Pharmacother, 2021, 142: 111533. doi: 10.1016/j.biopha.2021.111533
[18] Jang BI, Jung JY, Koh SA, et al. The important role of GPX1 and NF-κB signaling pathway in human gastric cancer: implications for cell proliferation and invasion [J]. Cancer Genomics Proteomics, 2024, 21(3): 305-315.
[19] Pang B, Mao H, Wang J, et al. MiR-185-5p suppresses acute myeloid leukemia by inhibiting GPX1 [J]. Microvasc Res, 2022, 140: 104296. doi: 10.1016/j.mvr.2021.104296
[20] Yuan X, Liu Y, Chen E, et al. MiR-646 regulates proliferation and migration of laryngeal carcinoma through the PI3K/AKT pathway via targeting GPX1 [J]. Oral Dis, 2021, 27(7): 1678-1686.
[21] Huergo C, Tornín J, Estuplñán O, et al. GPX1 expre-ssion promotes stemness and aggressiveness in myxoid liposarcomas [J]. Int J Biol Sci, 2025, 21(13): 5609-5627. doi: 10.7150/ijbs.105217
[22] Metere A, Frezzottlf F, Graves CE, et al. A possible role for selenoprotein glutathione peroxidase(GPx1)and thioredoxin reductases(TrxR1)in thyroid cancer: our experience in thyroid surgery [J]. Cancer Cell Int, 2018, 18: 7. doi: 10.1186/s12935-018-0504-4
[23] Liang X, Weng J, You Z, et al. Oxidative stress in cancer: from tumor and microenvironment remodeling to therapeutic frontiers [J]. Mol Cancer, 2025, 24(1): 219. doi: 10.1186/s12943-025-02375-x
[24] Son Y, Cheong YK, Kim NH, et al. Mitogen-activated protein kinases and reactive oxygen species: how can ROS activate MAPK pathways? [J]. J Signal Transduct, 2011, 2011: 792639. doi: 10.1155/2011/792639
[25] Guo YJ, Pan WW, Liu SB, et al. ERK/MAPK signa-lling pathway and tumorigenesis [J]. Exp Ther Med, 2020, 19(3): 1997-2007.
[26] Wang B, Wang Y, Zhang J, et al. ROS-induced lipid peroxidation modulates cell death outcome: mechanisms behind apoptosis, autophagy, and ferroptosis [J]. Arch Toxicol, 2023, 97(6): 1439-1451.
[27] 蒋崇辉, 艾小红, 杨小红, 等. 活性氧生成在8-硝基白杨素诱导人胃癌SGC-7901细胞凋亡中的作用 [J]. 山东大学学报(医学版), 2012, 50(2): 34-37. JIANG Chonghui, AI Xiaohong, YANG Xiaohong, et al. Generation of reactive oxygen species in apoptosis of human gastric cancer SGC-7901 cells induced by 5,7-dihydroxy-8-nitrochrysin [J]. Journal of Shandong University(Health Sciences), 2012, 50(2): 34-37.
[28] Hou Y, Wang H, Wu J, et al. Dissecting the pleiotropic roles of reactive oxygen species(ROS)in lung cancer: from carcinogenesis toward therapy [J]. Med Res Rev, 2024, 44(4): 1566-1595.

相关文章 15

[1]	张士宝刘庆勇阮喜云陈杰张建军李宗武杨广笑王全颖. NT₄-SAC-HA₂-TAT融合基因表达载体的构建及鉴定[J]. 山东大学学报(医学版), 2209, 47(6): 15-19.
[2]	王晓磊张海涛张辉郭成浩. 舒血宁注射液对高碘致培养血管内皮细胞损伤的保护作用[J]. 山东大学学报(医学版), 2209, 47(6): 38-.
[3]	鹿向东杨伟徐广明曲元明. 脑膜瘤中PPAR-γ的表达及曲格列酮对脑膜瘤培养细胞生长的影响[J]. 山东大学学报(医学版), 2209, 47(6): 65-.
[4]	韩觉明,王晖,吴倩,郑慧玲,朱琳. B4GALNT4促进肺腺癌细胞增殖、迁移和侵袭能力[J]. 山东大学学报 (医学版), 2025, 63(7): 23-31.
[5]	蔡佳莹,王靖婷,王增萍,王璟,郏雁飞,马晓丽. α5-nAChR对慢性应激肺腺癌荷瘤小鼠疑核c-Fos表达的影响[J]. 山东大学学报 (医学版), 2025, 63(4): 69-74.
[6]	李响,张艺,王雪纯,徐梦超,王月兰. 氧化应激在创伤性脑损伤诱发急性肺损伤中的研究进展[J]. 山东大学学报 (医学版), 2025, 63(2): 118-124.
[7]	李观强,施昱诚,朱可涵,胡波,黄献琛,孙元,李笃信,张喜成. 蜗牛粘液来源的活性肽SK-14促进成纤维细胞的增殖和迁移[J]. 山东大学学报 (医学版), 2025, 63(11): 1-7.
[8]	刘振昆,吕纪玲,徐伟伟,马力天,张才擎. BALF tNGS检测及培养对NSCLC合并IPFD的诊断价值[J]. 山东大学学报 (医学版), 2025, 63(11): 36-45.
[9]	张洁,张芳芳,王靖楠,李泽宇,宋颖,李娜. circ_0000144在乳腺癌中的表达及其对乳腺癌细胞增殖、迁移和侵袭能力的影响[J]. 山东大学学报 (医学版), 2025, 63(1): 35-42.
[10]	杜学识,倪向敏,梁馨予,白倩,朱文艺,王建. 雌马酚对DN的保护作用及潜在靶点[J]. 山东大学学报 (医学版), 2024, 62(8): 49-58.
[11]	王静,刘晓菲,曾荣,许长娟,张锦涛,董亮. 基于机器学习算法鉴定哮喘的坏死性凋亡相关生物标志物[J]. 山东大学学报 (医学版), 2024, 62(7): 21-32.
[12]	张学宇,张学海,孙文青,刘晗,姜金波,刘寒,李远,陈晓梅. 重症新型冠状病毒肺炎伴侵袭性肺曲霉病及反复致命性消化道出血1例[J]. 山东大学学报 (医学版), 2024, 62(7): 56-61.
[13]	王靖婷,王璟,鲁艺,李静坦,李强,郏雁飞,马晓丽. α5-nAChR与MHC-I在肺腺癌中的表达及相关性[J]. 山东大学学报 (医学版), 2024, 62(5): 72-78.
[14]	姜子晗,芦兴晨,孙露,赵蕙琛,左丹,马小莉,刘元涛,张玉超. NR4A1通过IκBα/NF-κB通路调控过氧化氢诱导人脐静脉内皮细胞凋亡的机制[J]. 山东大学学报 (医学版), 2024, 62(3): 11-19.
[15]	杨闯,张荣雨,宋彬,王程君,赵文,玄甜甜,李际盛. 阿美替尼一线治疗EGFR突变肺腺癌伴大疱性类天疱疮1例并文献复习[J]. 山东大学学报 (医学版), 2024, 62(12): 32-37.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed