Fig. 5: COL4A6 regulates cell sensitivity to cisplatin via the DDR1/NF-κB axis. | Oncogenesis

Fig. 5: COL4A6 regulates cell sensitivity to cisplatin via the DDR1/NF-κB axis.

From: Collagen type IV alpha 6 promotes tumor progression and chemoresistance in ovarian cancer by activating the discoidin domain receptor 1 pathway

Fig. 5

A Upper panel: A2780CP70 cells were transfected with COL4A6 knockdown plasmid. After 48 h, the cells were seeded into 96-well plates and treated with various concentrations of cisplatin for 48 h; subsequently, cell sensitivity to cisplatin was measured using the MTT assay. Lower panel: A2780 cells were transfected with COL4A6-expression plasmid. After 48 h, the cells were seeded into a 96-well plate and treated with various concentrations of cisplatin for 48 h; subsequently, cell sensitivity to cisplatin was measured using the MTT assay. All experiments were performed in triplicate. P-values were determined using the Student’s t-test. *P < 0.05 or **P < 0.005, relative to control cells treated with shControl or V. B A2780CP70 cells were transfected with COL4A6 knockdown plasmid and a DDR1 cDNA plasmid, and A2780 cells were transfected with a COL4A6 cDNA plasmid and DDR1 knockdown plasmid. COL4A6, DDR1, p-DDR1, p-IKKβ, p-IKKγ, p65, COX2, and the nuclear p65 fraction in whole lysates of both types of cells were evaluated by western blotting. β-actin and SP1 were detected as loading controls for whole-cell lysates and nuclear fractions, respectively. C A2780CP70 cells were transfected with different quantities of a COL4A6 knockdown plasmid (1 and 3 µg) and a DDR1 cDNA plasmid (3 µg), and A2780 cells were transfected with different quantities of a COL4A6 plasmid (1 and 3 µg) and DDR1 knockdown plasmid (3 µg). Luciferase activities were measured and normalized to Renilla luciferase activities. All experiments were performed in triplicate. P-values were determined using the Student’s t-test. *P < 0.05 or **P < 0.005, relative to control cells treated with shControl or V. D A2780CP70 cells were transfected with different quantities of a COL4A6 knockdown plasmid (1 and 3 µg) and a DDR1 cDNA plasmid (3 µg), and A2780 cells were transfected with different quantities of a COL4A6 plasmid (1 and 3 µg) and DDR1 knockdown plasmid (3 µg). After 48 h, the cells were seeded into a 96-well plate and treated with various concentrations of cisplatin for 48 h; subsequently, cell sensitivity to anticancer drugs was measured using the MTT assay. All experiments were performed in triplicate. P-values were determined using the Student’s t-test. *P < 0.05 or **P < 0.005, relative to control cells treated with shControl or V. E Upper panel: western blotting was performed to evaluate COL4A6, DDR1, p-DDR1, and COX2, and the nuclear p65 fraction in whole lysates of both types of cells was evaluated by western blotting. β-actin and SP1 served as loading controls for whole-cell lysates and nuclear fractions, respectively, after treatment of A2780-COL4A6 expression cells for 24 h with an NF-κB inhibitor. β-actin was the loading control. Lower panel: After 48 h, the cells were seeded into a 96-well plate and treated with various concentrations of cisplatin for 48 h; subsequently, cell sensitivity to anticancer drugs was measured using the MTT assay. All experiments were performed in triplicate. P values were determined using the Student’s t-test. *P < 0.05 or **P < 0.005, relative to control cells treated with shControl or V.

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