Escin (Aescin) is biosynthesized from protoaescigenin and barringtogenol, occurring in the seeds of Aesculus.

Escin is effective to inhibit pancreatic cancer cell proliferation as a single agent. However, escin has minimal effect on normal rat skeletal muscle cells. Escin potentiates the proliferation-inhibiting and apoptosis-inducing effect of gemcitabine in both BxPC-3 and PANC-1 cell lines in vitro. It also down-regulates constitutive as well as gemcitabine-induced activation of NF-kB and its downstream gene products in pancreatic cancer cells in vitro[1].

Escin augments therapeutic effect of gemcitabine in BxPC-3 xenografts in nude mice. They have significant synergistic effects on suppressing the growth of pancreatic tumors, inhibiting the proliferation and inducing apoptosis of tumors. Escin potentiates the inhibiting effect of gemcitabine on NF-kB and NF-kB-regulated gene products in BxPC-3 xenografts in nude mice[1].

Cell cycle analysis: After treatment with escin (BxPC-3 (25 μM),PANC-1 (20 μM),respectively), gemcitabine (BxPC-3(100 nM), PANC-1(1 μM), respectively) or their combinations for 72 h, BxPC-3 and PANC-1 cells are collected by trypsinization, washed with ice-cold PBS and fixed overnight by the addition of 2 ml of ice-cold 70% ethanol/30% PBS at 4°C. The ethanol is subsequently removed after centrifugation, and about 1×106 cells are resuspended in 800 μl of PBS, 100 μl of ribonuclease A (500 g/ml PBS) and 100 μl of PI (500 g/ml PBS) in the dark at room temperature for 30 min. Flow cytometric analysis is performed using FACScan for the detection of the percentage of cells in the diVerent phases of the cell cycle. Cells treated with DMSO alone are used as controls. (Only for Reference)

DMSO: 1 mg/mL (0.88 mM), Sonication is recommended.

H2O: < 1 mg/mL (insoluble or slightly soluble)

Ethanol: < 1 mg/mL (insoluble or slightly soluble)

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 1 mg/mL (0.88 mM), Sonication is recommended.