抑制NRF1/ABCC1提高人肺腺癌细胞系对顺铂化学治疗的敏感性  

Inhibition of NRF1/ABCC1 improves chemosensitivity of human lung adenocarcinoma cell lines to cisplatin

作　　者：陈清霞 梁玉玲[1] 罗亚兰 牛斌 CHEN Qingxia;LIANG Yuling;LUO Yalan;NIU Bin(Department of Clinical Laboratory,Chengdu Third People′s Hospital,Chengdu 610000,China)

机构地区：[1]成都市第三人民医院检验科,四川成都610000

出　　处：《基础医学与临床》2025年第1期51-59,共9页Basic and Clinical Medicine

基　　金：四川省医学(青年创新)科研课题(Q20009)。

摘　　要：目的探讨核呼吸因子1(NRF-1)激活ATP结合盒转运蛋白C1(ABCC1)转录对肺腺癌细胞顺铂抵抗的影响及其潜在机制。方法通过癌基因组数据集分析肺腺癌肿瘤组织中ABCC1的表达水平。细胞分组:sh-NC、sh-ABCC1、sh-NC+oe-NC、sh-NRF1+oe-NC和sh-NRF1+oe-ABCC1。RT-qPCR检测ABCC1在肺腺癌细胞中的表达量。构建顺铂耐药肺腺癌细胞株,检测ABCC1的表达水平。(0、0.001,0.002,0.004、0.008、0.016和0.032 mg/mL)顺铂处理的耐药肺腺癌细胞的IC50值。Western blot检测上皮间充质转化标志物(E-cadherin、N-cadherin)蛋白的表达。双荧光素酶报告基因和ChIP实验验证NRF1与ABCC1的结合关系。结果ABCC1在肺腺癌肿瘤组织和细胞中高表达。与顺铂敏感的肺腺癌细胞相比,ABCC1在顺铂耐药肺腺癌细胞中高表达,敲低ABCC1可显著抑制细胞增殖、迁移和侵袭,并提高E-cadherin的表达(P<0.05),降低N-cadherin的表达(P<0.05)。敲低ABCC1可显著提高肺腺癌细胞对顺铂的敏感性(P<0.05)。此外,双荧光素酶报告基因和ChIP实验证实NRF1与ABCC1启动子的去结合关系,且NRF1可激活ABCC1的转录。敲低NRF1可减弱过表达ABCC1对肺腺癌细胞增殖、迁移、侵袭及顺铂抵抗的抑制作用(P<0.05)。结论NRF1/ABCC1轴在肺腺癌顺铂抵抗中具有促进作用。抑制NRF1/ABCC1轴可能是提高肺腺癌顺铂敏感性的潜在靶点。Objective To explore the effect of nuclear respiratory factor 1(NRF1)activation of ATP binding cassette transporter C1(ABCC1)transcription on cisplatin resistance in lung adenocarcinoma cells and its potential mechanisms.Methods The expression levels of ABCC1 in lung adenocarcinoma tumor tissues were analyzed by oncogenomic datasets.Cells were grouped into:sh-NC,sh-ABCC1,sh-NC+oe-NC,sh-NRF1+oe-NC and sh-NRF1+oe-ABCC1.Real-time polymerase chain reaction(RT-qPCR)was used to detect the expression of ABCC1 in lung adenocarcinoma cells.Cisplatin-resistant lung adenocarcinoma cell strains were constructed to detect the expression level of ABCC1.Cell proliferation,migration and invasion were assessed by colony formation and Transwell assay.The IC 50 values of drug-resistant lung adenocarcinoma cells treated with different doses(0,0.001,0.002,0.004,0.008,0.016 and 0.032 mg/mL)of cisplatin were detected by CCK-8 assay.Western blot was used to detect the protein expression of epithelial-mesenchymal transition markers(E-cadherin,N-cadherin).Dual luciferase and ChIP assays were performed to verify the binding relationship between NRF1 and ABCC1.Results ABCC1 was highly expressed in lung adenocarcinoma tumor tissues and cells.Compared with cisplatin-sensitive lung adenocarcinoma cells,ABCC1 was highly expressed in cisplatin-resistant lung adenocarcinoma cells,and knockdown of ABCC1 significantly inhibited cell proliferation,migration and invasion,and increased the expression of E-cadherin(P<0.05)and decreased the expression of N-cadherin(P<0.05).Knockdown of ABCC1 significantly increased the sensitivity of lung adenocarcinoma cells to cisplatin(P<0.05).In addition,dual luciferase and ChIP experiments confirmed the binding relationship between NRF1 and ABCC1 promoter de-binding,and NRF1 could activate the transcription of ABCC1.Knockdown of NRF1 attenuated the inhibitory effect of over-expressed ABCC1 on the proliferation,migration,invasion and cisplatin-resistance of lung adenocarcinoma cells(P<0.05).Conclusions This study re

关 键 词：核呼吸因子(NRF1) ATP结合盒转运蛋白C1(ABCC1) 肺腺癌 上皮细胞-间充质转化 化学治疗抵抗 

分 类 号：R734.2[医药卫生—肿瘤]