Scutellaria baicalensis extract (SbE) has been shown to exert chemopreventive effects on several types of
cancer.
Baicalin, a hydrophilic
flavonoid found in SbE, may have opposing effects that decrease the antitumor potential of SbE against
colorectal cancer. In this study, after removing
baicalin, we prepared an aglycone-rich fraction (ARF) of SbE and evaluated its anti-proliferative activity and mechanisms of action. The
flavonoids found in ARF,
baicalin fraction (BF) and SbE were determined by high-performance liquid chromatography (HPLC). The effects of ARF, BF, SbE and representative
flavonoids on the proliferation of HCT-116 and HT-29 human
colorectal cancer cells were determined by an MTS assay. The cell cycle, the expression of
cyclins A and B1 and cell apoptosis were assayed using flow cytometry. Apoptosis-related gene expression was visualized by quantitative real-time polymerase chain reaction (PCR), and mitochondrial membrane potential was estimated following staining with
JC-1. HPLC analysis showed that ARF contained two hydrophobic
flavonoids,
baicalein and
wogonin, and that BF contained only
baicalin. SbE had little anti-proliferative effect on the
colorectal cancer cells;
cancer cell growth was even observed at certain concentrations. ARF exerted potent anti-proliferative effects on the
cancer cells. By contrast, BF increased
cancer cell growth. ARF arrested cells in the S and G2/M phases, increased the expression of
cyclins A and B1, and significantly induced cell apoptosis. Multiple genes in the mitochondrial pathway are involved in ARF-induced apoptosis, and subsequent cellular functional analysis validated the involvement of this pathway. These results suggest that removing
baicalin from SbE produces an ARF that significantly inhibits the growth of
colorectal cancer cells, and that the mitochondrial apoptotic pathway plays a role in hydrophobic
flavonoid-induced apoptosis.