Rab26 enhances the sensitivity of lung cancer cells resistant to Osimertinib by negative regulation of Nrf2

doi:10.3877/cma.j.issn.1674-6902.2024.03.002

Abstract

Abstract:

Objective

To explore the expression of Rab26 in osimertinib-resistant cells, and the role and mechanism of regulating lung cancer resistant cells′sensitivity to osimertinib.

Methods

Using a concentration gradient increasing method, induce H1975 cells to become osimertinib-resistant cell line (H1975OR). Western blot and qPCR were utilized to detect the expression levels of Rab26 and Nrf2 mRNA and proteins in H1975 and H1975OR. After overexpressing Rab26 through lentiviral infection in H1975OR cells (Rab26 OE group), continue to treat them with 1 μM osimertinib for 48 hours. Assess cell viability and apoptosis through CCK-8 and TUNEL, and evaluate ROS levels through flow cytometry. Use Western blot to assess Nrf2 protein expression levels in H1975 and H1975OR after treatment with 10 mM MG132 for 8 hours. Transfect H1975OR cells with Nrf2 siRNA, followed by treatment with 1 μM osimertinib for 48 hours, and evaluate cell viability using CCK-8 and ROS levels through flow cytometry.

Results

Rab26 mRNA and protein expression in H1975OR were significantly lower than in H1975 cells (P<0.05), while Nrf2 protein expression in H1975OR was significantly higher than in H1975 cells (P<0.05), with no significant difference at the mRNA level between the two groups (P>0.05). After osimertinib intervention, the cell viability of the Rab26 OE group was significantly lower than the control group (Vector), with a significant increase in apoptosis rate and ROS levels compared to the Vector group (P<0.05). The expression level of Nrf2 protein in the Rab26 OE group was significantly lower than in the Vector group, but there was no significant difference in Nrf2 mRNA between the two groups (P>0.05). After MG132 treatment, the Nrf2 protein level significantly increased in both the Vector group and Rab26 OE group (P>0.05). Knocking down Nrf2 significantly increased ROS levels in H1975OR cells after osimertinib intervention, while inhibiting cell viability.

Conclusion

Rab26 may promote the oxidative stress response induced by osimertinib by negatively regulating Nrf2, enhancing sensitivity of lung cancer resistant cells to osimertinib.

Key words: Bronchogenic carcinoma, Rab26, Osimertinib resistance, Nrf2, Oxidative Stress

Guoqing Yin, Li Zeng, Binfeng He, Fenfen Sun. Rab26 enhances the sensitivity of lung cancer cells resistant to Osimertinib by negative regulation of Nrf2[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(03): 349-355.

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Figures/Tables 5

References 36

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