Fig. 1 | Journal of Cancer Research and Clinical Oncology

Fig. 1

From: Resveratrol induces PD-L1 expression through snail-driven activation of Wnt pathway in lung cancer cells

Fig. 1

Resveratrol activates Wnt pathway to upregulate PD-L1 expression. a Western blot demonstrates increased PD-L1 expression following 3 h of resveratrol (0, 1, 5, 10, 20 and 30 μM) stimulation in lung cancer cell lines H1299 (left panel) and the levels of PD-L1 was quantified by densitometry (right panel) (**p < 0.01, for difference from untreated control cells by ANOVA with Dunnett’s correction for multiple comparisons). b Resveratrol also increased PD-L1 expression in A549 and H460 cells. c Western blot demonstrates increased PD-L1 expression following 3 h of low-dose resveratrol (< 5 μM) stimulation but decreased PD-L1 expression following 3 h of high-dose resveratrol (> 40 μM) stimulation in H1299 cell line. d RNA was extracted and subjected to qRT-PCR in A549 and H1299 cells with indicated resveratrol treatment. Values represent the relative reduction of PD-L1 mRNA levels normalized to GAPDH. The bars represent the mean ± SD of triplicates (**p < 0.01, ***p < 0.001, ****p < 0.0001 for difference from untreated control cells by ANOVA with Dunnett’s correction for multiple comparisons). e A549 and H1299 cells were co-transfected with PD-L1 promoter and control Renilla luciferase reporter gene plasmid and treated with indicated resveratrol concentrations after 48 h. Luciferase activity was determined and normalized using the dual luciferase reporter system (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001 for the difference from the control cells by ANOVA with Dunnett’s correction for multiple comparisons). f Representative Western blot analysis of PD-L1 in H1299 cells pre-treated with FH535, GSK3β, LY294002, LY2157299 at a concentration of 2 μM for 1 h and then with Resveratrol for 3 h. g A549 and H1299 cells were transiently co-transfected with Wnt3A cDNA, along with the TOPflash luciferase reporter, and luciferase activity was determined after transfection for 48 h and normalized using the dual luciferase reporter system. The bars represent the mean ±sSD of triplicates (**p < 0.01, ****p < 0.0001 for difference from untreated control cells by ANOVA with Dunnett’s correction for multiple comparisons). h A549 and H1299 cells were transiently co-transfected with Wnt3A cDNA, along with the PD-L1 promoter reporter construct, and luciferase activity was determined and normalized after transfection for 48 h using the dual luciferase reporter system (***p < 0.001, ****p < 0.001 for the difference from the control cells by ANOVA with Dunnett’s correction for multiple comparisons). i Overexpression of Wnt3A increased PD-L1 expression in dose-dependent manner

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