Abstract
ABSTRACTHaberlea rhodopensis—a Balkan endemite belonging to the group of resurrection plants, was studied as a potential source of novel cancer modulating drugs. Human embryonic (HEK293 p53++/+) and prostate cancer cell lines—LNCaP (p53+/+) and PC3 (p53−/-) were used as a model to follow the reaction to oxidative and genotoxic stress after pre-treatment with H. rhodopensis extract. Oxidative stress was estimated by flow cytometry (FCS) andfluorescent plate reader (FPR) using reactive oxygen speciesfluorescent dye (H2DCFDA). UV induced DNA damage was assessed by FCS PUMA (p53 upregulated modulator of apoptosis) expression. Inflammatory pathways were challenged using synthetic peptidoglycan by FCS Act1 expression and NFkB reporter stable cell line. Pre-treated-cells were assessed using DCF dye (FPR and FCS) for ROS (Reactive Oxygen Species) generation after dose-dependent H2O2 stress and decreased signal compared to non-treated cells. H. rhodopensis had cumulative effect on cell death in PC3 cells. UV-inducedgenotoxic stress resulted in FCS-detected PUMA upregulation only in PC3. Haberlea treated cells had better vitality and their challenge using a bacterial peptidoglycan resulted in an upregulation of Act1, but only in LNCaP cells. The NFkB reporter vector revealed transcription factor activation upon treatment with only peptidoglycan, while Haberlea pre-treatment resulted in negative modulation of the NFkB induction.Our data show that H. rhodopensis extracts have anti-oxidative effect in cancer vs. normal cell lines and differentially modulate distinct cell lines in genotoxic and inflammatory stress, favoring NFKB activation in p53+/+ cells, while suppressing its signaling in p53−/- cells.
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