The growth inhibitory effect of EVn-50 or VB1 on cells was measured using the SRB assay as Skehan et al.
Cells were treated with various concentrations of EVn-50 or VB1 24 h.
The effect of EVn-50 or VB1 on inducing apoptosis of MDA-MB-435 cells was measured by TUNEL and DAPI staining.
The EVn-50 is composed 70% of lignans and VB1 and VB2 are two most abundant lignan in EVn-50 as revealed by HPLC analysis in Fig.
To dissect the underlying mechanisms of the anticancer effect exerted by EVn-50, we used breast cancer cell line MDA-MB-435 and liver cancer cell line SMMC-7721 to assess the influence of lignans mixture EVn-50 and the purified EVn-50 lignan compound VB1 on cancer cell cycle with flow cytometric analysis.
For this purpose, we examined the morphological changes of tubulin in cells treated with lignans mixture EVn-50 and its purified lignan compound VB1 by immunofluorescence staining.
Meanwhile, we observed that the nucleus of MDA-MB-435 cells treated with EVn-50 or VB1 exhibited condensed and cleaved morphology, a typical apoptotic characteristic (Elmore 2007), while the nucleus in control cells exhibited uniformity morphology (Fig.