Role of Wnt/β-catenin signaling pathway in miR-21-mediated cisplatin resistance in non-small cell lung cancer
- VernacularTitle:Wnt/β-catenin信号通路在miR-21介导的非小细胞肺癌顺铂耐药性中的作用
- Author:
Chuanhui ZHENG
1
;
Li LIN
;
Xia WANG
;
Naigang WANG
Author Information
- Publication Type:Journal Article
- Keywords: miR-21; non-small cell lung cancer; cisplatin; cisplatin resistance; PTEN; Wnt/β-catenin pathway
- From: Journal of Xi'an Jiaotong University(Medical Sciences) 2025;46(2):238-248
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the role and mechanism of microRNA-21-5p(miR-21)in cisplatin(CDDP)resistance in non-small cell lung cancer.Methods The expression of miR-21 in cancer tissues and paracancerous tissues of non-small cell lung cancer patients was detected by real-time fluorescence quantitative PCR.Non-small cell lung cancer CDDP-resistant cells H1299/CR and H1975/CR were constructed using non-small cell lung cancer cell lines H1299 and H1975.CCK-8 was used to detect the cell activity of each group of cells under CDDP treatment,and apoptosis was analyzed by flow cytometry.Dual luciferase was used to detect the role of miR-21 in relation to PTEN in H1299 cells,RT-qPCR was used to detect the miR-21 level,and Western blotting was used to detect the protein expression level of PTEN and PTEN downstream Wnt/β-catenin signaling pathway.H1299 and H1299/CR were used to construct a hormonal nude mouse model to verify the effect of miR-21 on the sensitivity of CDDP treatment in non-small cell lung cancer.Results The expression of miR-21 was significantly higher in cancer tissues than in adjacent normal tissues(P＜0.001).The expression of miR-21 in drug-resistant cells H1299/CR and H1975/CR was significantly elevated compared to that in H1299 and H1975 cells(P＜0.05).After transfection with miR-21 inhibitor,cell viability in the inhibitor group was significantly lower than in the inhibitor NC group when treated with CDDP ≥12 pmol/L(P＜0.001).Flow cytometry analysis showed that while the apoptosis rate in the CDDP and miR-21 inhibitor groups did not significantly differ from that in the untreated group,the apoptosis rate in the CDDP+miR-21 inhibitor group was significantly higher(P＜0.001).In contrast,after transfection with miR-21 mimic,the cell viability of the mimic-NC group was significantly reduced compared to the miR-21 mimic group when treated with CDDP ≥12 pmol/L(P＜0.05).TargetScan predicted that miR-21 could bind to the 3-UTR region of PTEN.Dual-luciferase reporter assays confirmed that miR-21 directly targeted PTEN.Overexpression of PTEN(PTEN-OE)together with miR-21 mimic co-transfection in H1299 cells resulted in decreased PTEN expression and increased levels of p-β-catenin(Ser552,Ser675)and β-catenin,while the PTEN expression in the miR-21 mimic+PTEN-OE group was elevated,with a corresponding decrease in p-β-catenin andβ-catenin levels.Flow cytometry showed that apoptosis was significantly increased in the mimic-NC group after CDDP treatment(P＜0.05).In H1299 xenograft models,after treatment with miR-21 mimic and CDDP,tumor growth was slower in the control+CDDP group than in the control group;tumors were larger in the miR-21+CDDP group than in the control+CDDP group at all time points.TUNEL staining revealed that apoptosis in the tumor tissues of the control+CDDP group was higher than in the control group,while apoptosis was reduced in the miR-21+CDDP group compared to the control+CDDP group.In the H1299/CR xenograft model,after 5 days of treatment,the tumors were smaller in the H1299/CR+CDDP group than in the H1299/CR group,and those in the H1299/CR+CDDP+inhibitor group were smaller than in the H1299/CR+CDDP group.TUNEL staining showed that apoptosis was increased in the H1299/CR+CDDP group compared to the H1299/CR group,and further increased in the H1299/CR+CDDP+inhibitor group.Conclusion In non-small cell lung cancer,miR-21 overexpression inhibits PTEN level and activates the Wnt/β-catenin pathway involved in the development of CDDP resistance in non-small cell lung cancer.Inhibition of miR-21 expression in tumor cells will enhance the sensitivity of tumor cells to CDDP.
