Induction of apoptosis by shikonin through coordinative modulation of the Bcl-2 family, p27, and p53, release of cytochrome c, and sequential activation of caspases in human colorectal carcinoma cells

被引:101
作者
Hsu, PC
Huang, YT
Tsai, ML
Wang, YJ
Lin, JK
Pan, MH [1 ]
机构
[1] Natl Kaohsiung Marine Univ, Dept Seafood Sci, Kaohsiung, Taiwan
[2] Natl Cheng Kung Univ, Coll Med, Dept Environm & Occupat Hlth, Tainan 70101, Taiwan
[3] Natl Taiwan Univ, Coll Med, Inst Biochem, Taipei 10764, Taiwan
[4] Natl Kaohsiung First Univ Sci & Technol, Dept Safety Hlth & Environm Engn, Kaohsiung, Taiwan
关键词
Shikonin; apoptosis; cytochrome c; caspase-9; caspase-3; poly-(ADP-ribose) polymerase; DNA fragmentation factor; caspase-activated deoxyribonuclease; Bcl-2; BCl-X-L;
D O I
10.1021/jf0495993
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
Shikonin is a main constituent of the roots of Lithospermum erythrorhizon that has antimutagenic activity. However, its other biological activities are not well-known. Shikonin displayed a strong inhibitory effect against human colorectal carcinoma COLO 205 cells and human leukemia HL-60 cells, with estimated IC50 values of 3.12 and 5.5 muM, respectively, but were less effective against human colorectal carcinoma HT-29 cells, with an estimated IC50 value of 14.8 muM. Induce apoptosis was confirmed in COLO 205 cells by DNA fragmentation and the appearance of a sub-G1 DNA peak, which were preceded by loss of mitochondrial membrane potential, reactive oxygen species (ROS) generation, cytochrome c release, and subsequent induction of pro-caspase-9 and -3 processing. Cleavages of poly(ADP-ribose) polymerase (PARP) and DNA fragmentation factor (DFF-45) were accompanied by activation of caspase-9 and -3 triggered by shikonin in COLO 205 cells. Here, we found that shikonin-induced apoptotic cell death was accompanied by upregulation of p27, p53, and Bad and down-regulation of Bcl-2 and Bcl-X-L, while shikonin had little effect on the levels of Bax protein. Taken together, we suggested that shikonin-induced apoptosis is triggered by the release of cytochrome c into cytosol, procaspase-9 processing, activation of caspase-3, degradation of PARP, and DNA fragmentation caused by the caspase-activated deoxyribonuclease through the digestion of DFF-45. The induction of apoptosis by shikonin may provide a pivotal mechanism for its cancer chemopreventive action.
引用
收藏
页码:6330 / 6337
页数:8
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