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