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Abstract
Purpose: To evaluate the effects of ten edible, medicinal Thai plant extracts on MCF-7 cell viability and cell migration, as well as their mechanism(s) of action.
 Methods: Ethanolic plant extracts of ten edible, medicinal plants were tested for their cytotoxicity against MCF-7 cells using sulforhodamine B (SRB). To investigate the cytotoxic mechanism(s) of action of these extracts, the study was examined gene expression and protein expression by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Cell migration was studied by wound healing assay.
 Results: Four of the ten test extracts were potently cytotoxic, Careya sphaerica (CS), Azadirachta indica (AI), Piper nigrum (PN) and Oroxylum indicum (OI) with half maximal inhibitory concentrations (IC50) less than 100 μg/mL. All four extracts stimulated ROS overgeneration, increased caspase 3 activity and decreased growth-related gene expression including cdk2, cdk4, cdk6, cyclin D1 and cyclin E. Furthermore, the extracts significantly enhanced cyclin-dependent kinase inhibitor (CDKI) p21 levels and activated cancer cell death. The four extracts, CS, AI, PN and OI, also significantly reduced cancer cell migration, with PN being the most potent.
 Conclusion: Extract of the edible plants CS, AI, PN and OI have in vitro anticancer activity and are promising starting points for the development of breast cancer drugs.
 Keywords: Careya sphaerica (CS), Azadirachta indica (AI), Piper nigrum (PN), Oroxylum indicum (OI), Breast cancer, Cell death
Highlights
Compounds derived from herbal medicines have long attracted consideration as a starting point for drug development including anticancer drug
Plant extract Oroxylum indicum (OI), by contrast induced a significant increase in the expression of all five gene
Different cdk isoforms have crucial roles in cancer cell growth via loss of regulation of the cell cycle, a hallmark feature of cancer. These results show that AZ, Careya sphaerica (CS) and Piper nigrum (PN) extracts tend to suppress cdk2 and cdk6, whereas OI extract significantly induces all of the CDKs
Summary
Compounds derived from herbal medicines have long attracted consideration as a starting point for drug development including anticancer drug. A wound was made in each MCF-7 cell monolayer using sterile 0.2 mL pipette tips These were incubated with different doses of extract (0 - 100 μg/mL) for 48 h. Cell was exposed to the various doses of extract (0 - 250 μg/mL) for 24 h, and caspase 3 activity was measured as described previously [1]. Cell were exposed to the various doses of extract (0 - 100 μg/mL) for 24 h, and gene expression (cdk, cdk, cdk, cyclin D1 and cyclin E) was measured by RT-PCR as described previously [1]. The final reaction volume (20 μL) consisted of target gene or internal control and Master Mix. Cells were exposed to a fixed doses of the extract (100 μg/mL) for 24 h, and protein expression was determined by Western blotting as described previously [1]. Statistical analysis was performed using the GraphPad Prism 5 program (GraphPad Software, San Diego, CA)
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