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
陈清霞 1梁玉玲 1罗亚兰 1牛斌1
作者信息
- 1. 成都市第三人民医院 检验科,四川 成都 610000
- 折叠
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.
Abstract
Objective To explore the effect of nuclear respiratory factor 1(NRF1)activation of ATP binding cas-sette 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 on-cogenomic 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 IC50 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 adenocarci-noma 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 experi-ments 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 revealed the effect of the NRF1/ABCC1 axis enhancement is a potential strategy to overcome the barrier of cisplatin-resistance in chemotherapy of lung adenocarcinoma.
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Key words
nuclear respiratory factor 1(NRF1)/ATP binding cassette transporter C1(ABCC1)/lung adenocarcinoma/epithelial-mesenchymal transition/chemotherapy resistance引用本文复制引用
出版年
2025
国家科技期刊平台
NSTL
万方数据