Bcl-2 degradation is an additional pro-apoptotic effect of polo-like kinase inhibition in cholangiocarcinoma cells

Figure 3 Impact of treatment with BI6727 and cisplatin on Bax and Bcl-2 expression in cholangiocellular carcinoma cell lines. Bax expression levels were determined in KMCH-1 (A-C) and Mz-Ch-1 (D-F) cells after treatment with BI6727 (200 nmol/L for 24 h), cisplatin (1 mmol/L for 24 h) or the combination of both components. Bax mRNA levels were measured by qRT-PCR (A, D; mean ± SEM, n = 3), shown as foldchange compared to vehicle-treated cells. Representative Western blot images of Bax and the loading control actin are shown in panels B and E. Bcl-2 protein levels were determined using Western blot analysis in treated Mz-Ch-1 cells. Proteasomal degradation was inhibited by co-treatment with MG-132 (1 μmol/L for 24 h) (G, H). Protein levels were quantified by densitometry, normalized to the loading control and shown as foldchange compared to vehicle-treated cells (C, F, H, mean ± SEM, n = 3); P-values were determined using student t-test. CCA: Cholangiocarcinoma; Bcl-2; B-cell lymphoma 2.