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Fig. 5 | Breast Cancer Research

Fig. 5

From: Metformin overcomes resistance to cisplatin in triple-negative breast cancer (TNBC) cells by targeting RAD51

Fig. 5

Metformin regulates RAD51 expression through the ERK pathway. a, b Hs 578T and MDA-MB-231 cells were treated with a 0.5~10 μM cisplatin and cultured for 6 h or b 5 μM cisplatin and cultured for the indicated times. Subsequently, 30-μg samples of whole cell lysates were subjected to western blotting using an antibody against phospho-ERK1/2 (Thr202/Tyr204) or total ERK1/2 (control). Bar graphs show immunoblotting band intensities. c, d Hs 578T and MDA-MB-231 cells were treated with metformin (1~10 mM) for 6 h (c) or with metformin (5 mM) for 0, 3, 6, 12, or 24 h (d). Whole cell lysates were subjected to immunoblotting for phospho-ERK1/2 (Thr202/Tyr204) and total ERK1/2. e Hs 578T and MDA-MB-231 cells were treated for 6 h with metformin (5 mM) and cisplatin (5 μM), either alone or in combination. Whole cell lysates were subjected to immunoblotting for phospho-ERK1/2 (Thr202/Tyr204) and total ERK1/2. f Hs 578T (upper panel) and MDA-MB-231 (lower panel) cells were treated with cisplatin (5 μM) for 24 h after pretreatment for 30 min with 30 μM PD98059, an ERK-specific inhibitor. Whole cell lysates were subjected to immunoblotting for phospho-ERK1/2 (Thr202/Tyr204), RAD51, total ERK1/2, or β-actin (loading control). g Hs 578T (left panel) and MDA-MB-231 (right panel) cells were pretreated with PD98059 (30 μM) for 30 min and then incubated with cisplatin (5 μM) for 24 h. Cell viability was determined by MTT assay. Results represent the mean ± SEM of five independent experiments. #, ## vs. no treatment; #P < 0.05, ##P < 0.01, *P < 0.05, **P < 0.01 by one-way ANOVA followed by Bonferroni’s post hoc test

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