| Carvacrol, a monoterpene phenol, present as an active constituent of the volatile oil of oregano and thyme, has been shown to have anti-proliferative effects on various human cancer cell lines, such as breast cancer, hepatoma, and cervical cancer. Arunasree K. showed that the anti-proliferative effects of carvacrol in metastatic breast cancer cells (MDA-MB-231) was based on the activation of the classical apoptosis response. However, its underlying mechanism of anti-proliferation on human metastatic breast cancer cells has not yet been determined. Also effect of carvacrol on angiogenesis and metastasis has never been studied. The objective of this research was to investigate the underlying mechanism of anti-proliferative effects of carvacrol on an estrogen insensitive, human, metastatic breast cancer cell line, MDA-MB-231 and also check effect of carvacrol on two additional aspects of cancer angiogenesis and metastatsis. In this study, cells were treated with increasing concentrations of carvacrol, ranging from 100 μM to 600 μM. Cell viability was determined after 24 h using the XTT cell proliferation assay. Carvacrol inhibited viability and resultant IC50 value was found to be 305.5811±14.875.Considering the IC50 to be close to 300 μM, we did further studies with inhibitor using carvacrol 300 μM in presence of specific inhibitors, by XTT cell proliferation assay. Western blot to check effect of carvacrol on NFKB translocation was also carried out using 300 μM concentration of carvacrol, and phosphorylation levels of ERK1/2 was checked by using three different concentrations of carvacrol 100, 300 and 600 μM. Our findings suggested that carvacrol induced anti-proliferative effect is independent of mitogen activated protein kinase pathway (MAPK) based on cell proliferation assay (24 h treatment). Phosphoinositol-3-phosphate pathway also did not 2 play an important role in anti-proliferative effect of carvacrol. Further investigations suggested that carvacrols anti-proliferative action is independent of NF-KB translocation. Phosphorylation of ERK1/2 increased at 600 μM, which may be due to non-specificity of drug at such a high concentration. But further investigation is suggested by performing time-dependent studies.;Interestingly, this study presents the first evidence of the anti-metastatic and antiangiogenic effect of carvacrol on MDA-MB-231 cells. The induction of anti-angiogenic effect by carvacrol was evident from decrease in vascular endothelial growth factor levels. The anti-metastatic potential of carvacrol was confirmed by decrease in cell migration of MDA-MB-231. Taken together, these findings suggest that carvacrol has anti-cancer potential in metastatic breast cancer. |
