Fig. 6: DDR1-IN-1 increases cell sensitivity to cisplatin and synergizes with cisplatin-mediated cell apoptosis via COL4A6 inhibition. | Oncogenesis

Fig. 6: DDR1-IN-1 increases cell sensitivity to cisplatin and synergizes with cisplatin-mediated cell apoptosis via COL4A6 inhibition.

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

Fig. 6

A Protein expression levels of COL4A6, DDR1, p-DDR1, SHC1, FAK, and p-FAK in A2780/V and A2780/COL4A6 cells treated with different concentrations of DDR1-IN-1 for 24 h were evaluated by western blotting. β-actin was used as an internal loading control. All experiments were performed in triplicate. B Invasion activity in vitro of A2780/V and A2780/COL4A6 cells after treatment with different concentrations of DDR1-IN-1 for 24 h. All data represent the means ± standard deviations of three separate experiments. *P < 0.05 and **P < 0.005, SC66 versus control. C Ovarian cancer cells were treated with different concentrations of cisplatin (0–30 μM) or combined with different concentrations of DDR1-IN-1 (0–20 μM) for 48 h. Each combination was tested with n = 5 replicates. After 48 h, cell viability was assessed by MTT assays. All experiments were performed in triplicate. The IC50 values of each agent alone or in combination treatments and CI values of the DDR1-IN-1 + CDDP combinations. P-value between the IC50 values of single versus combination treatment. D A2780/V and A2780/COL4A6 cells were treated for 24 h with 5 μM DDR1-IN-1 alone or with the addition of cisplatin (10 μM) indicated. The percentage of apoptotic cells was determined by annexin V and 7-AAD staining. The mean ± standard deviation for three independent experiments is shown. **P < 0.005, DDR1-IN-1 + CDDP versus CDDP. E Colony formation assay. A2780/V and A2780/COL4A6 cells were treated with 5 μM DDR1-IN-1 alone or with the addition of CDDP (10 μM) as indicated for 14 days. After treatment, the cells were stained with crystal violet. The mean ± standard deviation for three independent experiments is shown. **P < 0.005, DDR1-IN-1 + CDDP versus CDDP. F Western blotting was performed to evaluate COL4A6, DDR1, p-DDR1, SHC1, FAK, p-FAK, p65, COX2, and the nuclear p65 fraction in whole lysates of both types of A2780-COL4A6 expression cells after treatment with 5 μM DDR1-IN-1 alone or with the addition of CDDP (10 μM) for 24 h. β-actin and SP1 served as loading controls for whole-cell lysates and nuclear fractions, respectively. β-actin was the loading control.

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