Bcl-2 gene regulation

Melanoma is the least common but most dangerous skin cancer with the highest morbidity rate. Owing to increased toxicity towards normal cells, cancer chemo therapy is found to be great challenge with chemo sensitive agents. Phytochemicals derived anticancer agents is a field of great application. The unexplored plants remain the lead for novel therapeutic agents. Tylophoraindicaattracts research interest since little scientific data is available regarding the anticancer potential of the plant. In the present study purified ethanolic fraction showed the presence of compounds like alkaloids and quinines and the derivatives were confirmed by GC-MS analysis. Antiproliferative property of the alcoholic fraction was confirmed on malignant skin melanoma cell line SKMEL-28 by performing MTT assay, Neutral red uptake and LDH leakage assays respectively. The relative genotoxic effect of Tylophoraindica showed little damaging effects to genomic DNA when determined by comet assay and DNA fragmentation analysis. The apoptotic changes in nucleus were confirmed by fluorescent microscopic analysis by acridine orange/ethidium bromide staining. The molecular mechanisms of anticancer activity were mainly found to be due to the inhibition of BCL-2 gene expression by RT-PCR analysis. This study confirms anticancer activity of alcoholic fraction of Tylophoraindica, drawing new insights in their application for therapeutic regimens.