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[Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines]
Xia Fang1, Pan Gu, Cai-cun Zhou
1Department of Pathology, Shanghai Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
Objective:
To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms.
Methods:
PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin. β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.
Results:
Treating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations.
Conclusion:
Suppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.
Insights
Suppression of Wnt signaling can partially reverse gefitinib resistance in non-small cell lung cancer (NSCLC) cells. This approach may offer new therapeutic strategies for overcoming drug resistance in NSCLC patients.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Non-small cell lung cancer (NSCLC) often develops resistance to targeted therapies like gefitinib.
- The Wnt signaling pathway plays a role in cancer progression and drug resistance.
Purpose of the Study:
- To investigate the impact of inhibiting Wnt signaling on gefitinib resistance in NSCLC cells.
- To elucidate the underlying mechanisms of Wnt signaling's role in gefitinib resistance.
Main Methods:
- Gefitinib treatment of sensitive (PC9) and resistant (PC9/AB2) NSCLC cells.
- Assessment of cell proliferation using CCK8 assay.
- Analysis of Wnt signaling pathway members and beta-catenin transcriptional activity via Western blot and TOP Flash assay.
- Suppression of Wnt signaling using beta-catenin siRNA followed by gefitinib treatment and analysis of EGFR signaling.
Main Results:
- PC9/AB2 cells exhibited increased gefitinib resistance compared to PC9 cells.
- Elevated Wnt signaling pathway members and beta-catenin activity were observed in resistant cells.
- Wnt signaling suppression via siRNA increased apoptosis and gefitinib sensitivity.
- Downregulation of p-ERK1/2 was observed after Wnt signaling suppression.
Conclusions:
- Inhibition of Wnt signaling can partially restore sensitivity to gefitinib in NSCLC.
- Targeting the Wnt pathway presents a potential strategy to overcome gefitinib resistance in NSCLC.
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