Table 1.
Breast cancer cell lines used in HTS and their molecular classifications [4], [38].
Table 2.
Screen assay protocol for step 1 and 2.
Table 3.
FP assay performance and post-screen analysis summary.
Figure 1.
In vitro antitumorigenic activity of peonidin-3-glucoside or cyaniding-3-glucoside.
(A). Structures of hit compounds. (B) Effect of peonidin-3-glucoside and cyanidin-3-glucoside on HER2 and downstream signaling. BT474, MDA-MB-453, and HCC1569 cells were treated with or without peonidin-3-glucoside or cyanidin-3-glucoside for 6 h and whole-cell extracts were analyzed by western blotting with the indicated antibodies. (C) Peonidin-3-glucoside and cyanidin-3-glucoside treatments induce apoptosis in MDA-MB-453, BT474, and HCC1569 cells. Cells were treated with or without peonidin-3-glucoside or cyanidin-3-glucoside for 48 h and annexin V positive cells were counted using flow cytometry. (D) Peonidin-3-glucoside and cyanidin-3-glucoside treatments increase caspase 3/7 activity in MDA-MB-453, BT474, and HCC1569 cells. Cells were treated with or without peonidin-3-glucoside or cyanidin-3-glucoside for 48 h and cell viability and caspase 3/7 activity was measured as per the manufacturer's instructions. Data were normalized as caspase 3/7 activity divided by cell viability. Data represent mean ± SEM. *p<0.05, **p<0.01.
Table 4.
Summary of hit compounds.
Figure 2.
Antitumorigenic activity of peonidin-3-glucoside or cyaniding-3-glucoside in xenografted nude mice.
(A) Body weight of animals. (B) H&E staining for kidney, liver, and spleen. (C and D) Effects on tumor volume and tumor weight. Nude mice bearing MDA-MB-453 cells as xenografts were treated with control (saline), or group 1 (peonidin-3-glucoside (6 mg/kg/day)) or group 2 (cyaniding-3-glucoside (6 mg/kg/day)). Values are means ± SE (n = 10). *P<0.05. (E) H & E staining, expression of phospho-HER2 and Ki67. Control: saline; Group 1: peonidin-3-glucoside (6 mg/kg/day); group 2: cyaniding-3-glucoside (6 mg/kg/day).