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Fig. 5 | BMC Cancer

Fig. 5

From: DKK1 is a potential novel mediator of cisplatin-refractoriness in non-small cell lung cancer cell lines

Fig. 5

Knockdown of DKK1 sensitizes NSCLC cells to cisplatin. (a) Q-RT-PCR shows the relative mRNA expression of DKK1 48 and 72 h after a short pulse treatment of U-1810 cells with 10 μM cisplatin (1 h). The results of two separate experiments are shown. b U-1810 cells were transfected with non-targeting (NT) or DKK1-specific siRNAs (si1 or si2); silencing of DKK1 mRNA expression was confirmed by q-RT-PCR. 18S rRNA was used as a loading control. ***; p<0.005 vs NT control. c U-1810 cells were transfected with siRNA-DKK1 or NT siRNA and colony formation capacity was assayed 9 days after treatment or not with a short pulse of cisplatin (1 h, 10 μM). Representative images of culture dishes for si1. d Clonogenic capacity relative to untreated, NT siRNA-transfected cells for U-1810 si1 and si2. ***; p<0.005, **; p<0.01, when comparing DKK1 siRNA-transfected to NT siRNA-transfected cells after cisplatin treatment. e Western blots showing p21WAF1/Cip1, phosphorylated GSK3B (Ser9) and AKT (Ser473), total AKT, PI3K and Bcl-2 in U-1810 cells 24 h after treatment with cisplatin (1 h, 10 μM), which was performed on reseeded cells after the 72 h-transfection with NT control or siRNA-DKK1 (si1). GAPDH was used as a loading control. (f-g) A2780 and its cisplatin-resistant subtype A2780 cis cells were transfected with siRNA-DKK1 (si2) or NT. f siRNA silencing of DKK1 mRNA expression was confirmed by q-RT-PCR. **; p<0.01. g Cell viability was assayed 72 h after continuous treatment with cisplatin, relative to untreated NT or si, using MTT assay. *; p<0.05

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