Ginsenoside Rh4

Cell experiment:

Cell viability is measured via the CCK-8 assay. For these assays, Caco-2, HCT116 and CCD-18Co cells are seeded onto 96-well plates at a density of 1×104 cells per well and incubated overnight, followed by treatment with 0.1% (vol/vol) DMSO as vehicle or Ginsenoside Rh4 (60, 120, 180, 240 and 300 μM) for 24 or 48 h. Subsequently, the cells are incubated with 10% (vol/vol) the CCK-8 solution for 2 h. The absorbance at 450 nm is then read with a microplate reader. Cell viability is calculated[1].

Animal experiment:

Mice[1]Four-week-old female nude mice (12±2 g) are used. After acclimation for one week, the mice are inoculated with Caco-2 cells (1×107 cells/each) in the left forelimb pit. When the tumors reach 100 mm3, the mice are randomized into four groups (n=5): a control group (solvent), two Ginsenoside Rh4-treated groups (20 and 40 mg/kg/d) , and a positive control CAMPTO-treated group (40 mg/kg/3 d). The body weights and tumor sizes of the mice are measured every three days, and the tumor volume is calculated. After 30 days of treatment, the mice are sacrificed, and the xenograft tumors are removed and partially lysed to analyze the expression of cleaved-caspase 3, LC3, p-JNK and p-p53.

References:

[1]. Wu Q, et al. Ginsenoside Rh4 induces apoptosis and autophagic cell death through activation of the ROS/JNK/p53 pathway in colorectal cancer cells. Biochem Pharmacol. 2018 Feb;148:64-74.