The present invention relates to an anticancer composition use for cure or treatment of cancerous tumors, particularly human breast cancer. The natural compound Dentatin (DTN), or the use of a dentatin derivative (synthetic), using a pharmaceutically acceptable carrier for treatment of breast cancer. DTN could be useful as an adjunct theraphy for breast cancer.
Clausena excavata Burm. f. has been used in folk medicines in eastern Thailand for treatment of cancer. Dentatin (DTN) isolated from this plant, using a developed bioassay-guided approach, is further investigated in the probable apoptosis induction mechanism towards human breast cancer cells, MCF-7. DTN-induced cytotoxicity was observed with the MTT assay. Acridine orange/propidium iodide staining was used to detect cells in early apoptosis and high content screening (HCS) to observe nuclear condensation, cell permeability, mitochondrial membrane potential (MMP) and cytochrome c release.
Apoptosis induction was confirmed using clonogenic assay, DNA laddering and caspase 3/7 and 9 assays analysis. Reactive oxygen species (ROS) formation, Bcl-2 and Bax expression, and cell cycle arrest were also investigated. The involvement of nuclear factor-kappa B (NF-kB) was analysed with the HCS assay.
Our results demonstrated significant increased (p < 0.05) in chromatin condensation in cell nucleus observed by fluorescence analysis. Apoptosis induction was confirmed by the reduced number of colonies in the clonogenic assay and subsequent increased of cellular DNA breaks in treated MCF-7 cells observed as DNA ladder. Treatment of the MCF-7 cells with DTN encouraged apoptosis with cell death-transducing signals, which reduced MMP by down-regulation of Bcl-2 and up-regulation of Bax, triggering cytochrome c release from the mitochondria into the cytosol. Cytochrome c release triggered caspase 9 activation, followed by the executioner caspase 3/7. DTN treatment significantly arrested MCF-7 cells at G0/G1 phase (p