Previous work has shown alcoholic extracts of powdered ginger root to be cytotoxic to murine 4T1 breast cancer cells grown in culture, likely via induction of apoptosis (programmed cell death). In this study, the purification and identification of the cytotoxic compounds in ginger root were undertaken. Powdered ginger root was subjected to acetone reflux, and the resulting extract resolved into its component compounds by two rounds of thin layer chromatography (TLC). Three biologically active compounds (Bands 5, 6 and 9) were isolated and analyzed by high performance liquid chromatography (HPLC) and TLC in attempts to identify these compounds. The biological activity of the compounds was evaluated by bioluminescent cell viability assays. Additionally, 4T1 cells were treated in culture (24 hours) with the isolated compounds, and the effects on cellular protein expression were evaluated via one-dimensional polyacrylamide gel electrophoresis and Western blot analysis utilizing monoclonal antibodies specific for the caspase family of proteins, which mediate apoptosis. Results of the Western blot analysis supported a caspase-dependent mechanism of cell death. The involvement of an apoptotic pathway was confirmed by two-dimensional electrophoretic analysis of the cellular proteins after treatment with the purified compounds. HPLC and TLC analysis of the purified compounds showed that Band 5 co-migrated with 6-gingerol, and Band 9 co-migrated with 6-shogaol. Band 6 was chromatographically distinct and did not co-migrate with tested standards that are reported in the literature to cause cell death in 4T1 cells. Data suggest that Band 5 is 6-gingerol, Band 9 is 6-shogaol and Band 6 is likely a derivative of 6-shogaol. Further studies to characterize Bands 5, 6 and 9 are ongoing.

Purification and partial characterization of 4T1 cell cytotoxins from ginger (<i>Zingiber officinale</i>)

247th National Meeting (2014)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.

Purification and partial characterization of 4T1 cell cytotoxins from ginger (Zingiber officinale)