Effects of Crude Extracts from Medicinal Herbs Rhazya stricta and Zingiber officinale on Growth and Proliferation of Human Brain Cancer Cell Line In Vitro
Figure 3
Combination of CAERS and CFEZO induced an early biochemical feature of apoptosis. U251 cells were treated with indicated concentrations of CAERS and for 24 h and assayed for existence of apoptotic cell death. Depicted results are representative for independent experiments with almost identical observations. (a) DAPI staining showing combination of CAERS and CFEZO induced nuclear condensation, DNA fragmentation, and perinuclear apoptotic bodies in U251 cells (arrows). (b) Agarose gel showing CAERS and CFEZO induced DNA fragmentation in U251 cells. Lane “M” indicates the DNA marker ladder. (c) Comet assay showing formation of DNA tail in CAERS- and CFEZO-treated U251 cells. Nuclei with damaged DNA have the appearance of a Comet with a bright head and a tail, whereas nuclei with undamaged DNA appear round with no tail. In the panel denoted with 24 h, cells were treated with 20 μg CAERS and 20 μg CFEZO for 24 h; then treatment medium was discarded, cells were washed and grown in CAERS- and CFEZO-free medium for 24 before being harvested and assayed for comet analysis. The histogram displays percentage of cells with comet tail being analyzed in 50 cells for one slide. The bar denoted with 24, at the top, represents cells treated for 24 h, which then were grown in CAERS- and CFEZO-free medium for 24.
