Figure 5.

HIF-1α was an important effector in WDR5-induced CCA metastasis

(A) HIF1A mRNA in CCAs was significantly higher than that in normal tissues. (B and C) Quantitative real-time PCR (B) and western blot (C) show that the HIF-1α inhibitor LW6 (20 μM) or PX-478 (25 μM) decreased TWIST1 expression. (D) Survival curves of iCCA, pCCA, and dCCA patients were stratified by HIF-1α expression. (E) Genes in the HIF1A signaling pathway were changed along with WDR5 knockdown. (F and G) Invasive ability (F) and expression of EMT-specific proteins (G) in WDR5-overexpressed QBC939 cells in the presence or absence of LW6 (20 μM) or PX-478 (25 μM). (H) Metastatic models were established by tail vein injection of WDR5-overexpressed QBC939 cells in the treatment of HIF-1α inhibitor. Two HIF-1α inhibitors, LW6 (10 mg/kg p.o.) and PX-478 (100 mg/kg i.p.), were used to inhibit HIF-1α expression in vivo. The tumor metastases were monitored by a live imaging system (n = 6). (I) Radiant efficiency of in vivo fluorescence in (H) was measured to quantify the tumor burden of mice. (J) Livers of mice in (H) were weighed to assess the tumor burden of liver metastatic foci. (K and L) The numbers of metastasis nodules in livers (J) and lungs (K) of mice in (H) were counted. (M) The survival curves were further stratified into subgroups with the co-expression, single expression, and double low expression of WDR5 and HIF-1α in iCCA, pCCA and dCCA. In (B), (F), and (I)–(L), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, calculated by one-way ANOVA. In (D) and (M), data were calculated by a log-rank test, and the C-index was calculated to evaluate the accuracy of the prognostic model.