Induction of necrosis and apoptosis to KB cancer cells by sanguinarine is associated with reactive oxygen species production and mitochondrial membrane depolarization.
Sanguinarine is a benzopheanthridine alkaloid present in the root of Sanguinaria canadensis L. and Chellidonium majus L. In this study, sanguinarine (2 and 3 microM) exhibited cytotoxicity to KB cancer cells by decreasing MTT reduction to 83% and 52% of control after 24-h of exposure. Sanguinarine also inhibited the colony forming capacity (>52-58%) and growth of KB cancer cells at concentrations higher than 0.5-1 microM. Short-term exposure to sanguinarine (>0.5 microM) effectively suppressed the adhesion of KB cells to collagen and fibronectin (FN). Sanguinarine (2 and 3 microM) induced evident apoptosis as indicated by an increase in sub-G0/G1 populations, which was detected after 6-h of exposure. Only a slight increase in cells arresting in S-phase and G2/M was noted. Induction of KB cell apoptosis and necrosis by sanguinarine (2 and 3 microM) was further confirmed by Annexin V-PI dual staining flow cytometry and the presence of DNA fragmentation. The cytotoxicity by sanguinarine was accompanied by an increase in production of reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential as indicated by single cell flow cytometric analysis of DCF and rhodamine fluorescence. NAC (1 and 3 mM) and catalase (2000 U/ml) prevented the sanguinarine-induced ROS production and cytotoxicity, whereas dimethylthiourea (DMT) showed no marked preventive effect. These results suggest that sanguinarine has anticarcinogenic properties with induction of ROS production and mitochondrial membrane depolarization, which mediate cancer cell death.
Chang
Mei-Chi
MC
Biomedical Science Team, Chang Gung Institute of Technology, Taoyuan, Taiwan.
Chan
Chiu-Po
CP
Wang
Ying-Jan
YJ
Lee
Po-Hsuen
PH
Chen
Lin-I
LI
Tsai
Yi-Ling
YL
Lin
Bor-Ru
BR
Wang
Yin-Lin
YL
Jeng
Jiiang-Huei
JH
D000111:Acetylcysteine / Q000494:pharmacology; D000470:Alkaloids / Q000494:pharmacology*; D017304:Annexin A5 / Q000378:metabolism; D016588:Anticarcinogenic Agents; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053119:Benzophenanthridines / Q000494:pharmacology*; D002374:Catalase / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D005353:Fibronectins / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology*; D007624:KB Cells; D008564:Membrane Potentials / Q000187:drug effects; D051336:Mitochondrial Membranes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D009336:Necrosis; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D014410:Tumor Stem Cell Assay
D000470:Alkaloids; D017304:Annexin A5; D016588:Anticarcinogenic Agents; D000975:Antioxidants; D053119:Benzophenanthridines; D005353:Fibronectins; D007546:Isoquinolines; D017382:Reactive Oxygen Species; C038983:1,3-dimethylthiourea; D003094:Collagen; C005705:sanguinarine; D002374:Catalase; D013890:Thiourea; D000111:Acetylcysteine