Anti-proliferative Effects of Organic Extracts from Root Bark of Juglans Regia L. (RBJR) on MDA-MB-231 Human Breast Cancer Cells: Role of Bcl-2/Bax, Caspases and Tp53

With increasing use of plant-based cancer chemotherapeutic agents, exploring the antiproliferative effects ofphytochemicals has gained increasing momentum for anticancer drug design. The present study was undertakento investigate the effect of root bark of Juglans regia (RBJR) organic extracts on cell proliferation, and todetermine the molecular mechanism of RBJR-induced cell death by determining the expression of Bcl-2, Bax,caspases, Tp53, Mdm-2 and TNF-α in MDA-MB-231 human breast cancer cells. The results demonstrate thatWNRB suppressed proliferation and induced apoptosis in a dose and time dependent manner by modulatingexpression of key genes. This involved characteristic changes in cytoplasmic and nuclear morphology, DNAfragmentation (TUNEL assay), levels of mRNA and expression of corresponding proteins. Real Time PCR andwestern blot analysis revealed that the expression of of Bax, caspases, tp53, and TNF-α was markedly increasedin MBA-MB-231 cells treated with the RBJR extract. In contrast Bcl2 and mdm-2 expression was down regulatedafter exposure. In summary, our data suggest the presence of bioactive compound(s) in WNRB capable of killingbreast carcinoma cells through induction of apoptosis, and therefore a candidate source of anticancer drugs.