Figure 1.
XZH-5 reduces STAT3 phosphorylation.
(A) Structure of XZH-5. (B) MDA-MB-231, SUM 159, PANC-1, and SW1990 cells were treated with different concentrations of XZH-5 for 8 hours. p-STAT3 and STAT3 were analyzed by western blot. (C) MDA-MB-231, SUM 159, PANC-1 and SW1990 were treated with XZH-5 for 8 hours. The mRNA expression of BCL-2, BCL-xL, CyclinD1, Survivin, and GAPDH was analyzed by RT-PCR.
Figure 2.
(A) MDA-MB-231, SUM 159, PANC-1 and SW1990 cells were treated with XZH-5 for 8 hours. Cleaved PARP and Cleaved Caspase-3 were analyzed by western blot. (B) Caspase-3/7 activity was measured in XZH-5 treated MDA-MB-231, SUM 159, PANC-1 and SW1990 cells. The data represented three independent results.
Figure 3.
XZH-5 inhibits IL-6-induced STAT3 phosphorylation.
MCF-7 and ASPC-1 cells were pre-treated with XZH-5 for 2 hours, followed by 50 ng/ml of IL-6 (A) or IFN-γ (B). p-STAT3 (A) and p-STAT1 (B) were analyzed by western blot.
Figure 4.
XZH-5 blocks the IL-6-induced p-STAT3 nuclear accumulation and STAT3 nuclear translocation.
MCF-7 cells were pre-treated with 50 µM of XZH-5 for 2 hours followed by 50 ng/ml of IL-6 for 30 minutes. After the treatment, the localization of p-STAT3 (A) and STAT3 (B) was analyzed by immunofluorescence.
Figure 5.
XZH-5 reduces colony forming ability and inhibits cell migration.
(A) MDA-MB-231 and PANC-1 cells were treated with XZH-5 for 2 hours. After the treatment, viable cells were counted and the same number of cells were seeded and cultured for two week. Colonies were fixed by ice-cold methanol and were stained by 1% crystal violet. (B) When MDA-MB-231 and PANC-1 cells were 100% confluent, the monolayer was scratched using a pipette tip. Then the cells were treated with XZH-5 or DMSO for 2 hours. After the treatment, fresh medium was added and cells were cultured for 48 hours.
Figure 6.
XZH-5 enhances cytotoxicity of chemotherapeutic drugs when combined with other anti-cancer drugs.
(A) MDA-MB-231 cells were treated with 2.5 µM of Doxorubicin with or without 15 µM and 20 µM of XZH-5. (B) PANC-1 cells were treated with 250 nM of Gemcitabine with or without 15 µM and 20 µM of XZH-5. After 36 hours, cell viability was measured by fluorescence.