In vivo anti-tumor evaluation
All animal experiments were
approved by the Institutional Animal Care and Treatment Committee of
Chengdu University of TCM (Chengdu, P. R. China). Female Balb/c nude
mice and Balb/c mice (6-8 weeks old) were purchased from HFK Bioscience
CO., Ltd. (Beijing, China). These Balb/c mice or Balb/c nude mice were
inoculated with 100 μL of 4T1 cell suspension (1.0×106cells) or MDA-MB-231 cell suspension (1.0×107 cells).
At 5 days after inoculation, when the tumor volume had reached about 100
mm3, the mice were equally and randomly divided into
three treatment groups (control, 20
mg/kg, and 40 mg/kg) (n=6).
Mice
received icariin treatment by intraperitoneal injection once daily for
two weeks. The tumor volume (0.5 × L × W2, where L is
the length and W is the width) and body weight of the mice were recorded
every two days. The mice were
euthanized
by cervical dislocation at the termination of the experiments.
The tumors were isolated, imaged,
weighted, and fixed with
paraformaldehyde for further
immunohistochemistry
evaluation.