探討連翹萃取物對人類乳癌細胞 MCF-7 細胞死亡之作用機制

Abstract

Forsythia suspensa is a traditional Chinese medicine, which is a climbing plant widely distributed in Asia. It was reported to exhibit potential antibacterial, antiviral and anti-inflammatory. This study is to investigate the inhibition of human breast cancer cells MCF-7 proliferation by the extract of F. suspensa The murine macrophage cell line RAW264.7 stimulated with lipopolysaccharide (LPS) cocultured with methanol extractions of F. suspensa and showed significantly suppress the production of nitric oxide in a dose dependent manner. The MTT assay showed dose-dependent decrease in MCF-7 cell viability but not in normal cell. By confocal microscopy using PI staining also found the condensation and shrinkage of the cell nucleus. Flow cytometric analysis demonstrated that the F. suspensa extract increase sub-G1 population could induce apoptosis. In addition, western blot analysis showed that the F. suspensa extract could reduce Bcl-2 expression and activate PARP cleavage. It also suppressed p-Akt and p-ERK activation to block PI3K/AKT/mTOR and MAPK pathway by dose-dependent and time-dependent manner. F. suspensa extract coluld promote LC3B increase; moreover, AO staining was also found AVOs increased. This was accompanied by a decrease in cell viability due to activation of cell death by apoptosis and the occurrence of autophagy. The active components of F. suspensa extract caffeic acid ,forsythoside, forsythin and rutin were determined by HPLC. However, the underlying cytotoxicity phenomenon of F. suspensa extract have been elucidated in current study. Future investigation need to be done in the cytotoxicity mechanism of F. suspensa extract. This study implies that F. suspensa extract showing the potential of anti-tumor activity in breast cancer.