Author(s): Chen, TC (Chen, Ting-Ching); Lai, KC (Lai, Kuang-Chi); Yang, JS (Yang, Jai- Sing); Liao, CL (Liao, Ching-Lung); Hsia, TC (Hsia, Te-Chun); Chen, GW (Chen, Guang-Wei);
Lin, JJ (Lin, Jen-Jyh); Lin, HJ (Lin, Hui-Ju); Chiu, TH (Chiu, Tsan-Hung); Tang, YJ (Tang, Yih- Jing); Chung, JG (Chung, Jing-Gung)
Title: Involvement of reactive oxygen species and caspase-dependent pathway in berberine- induced cell cycle arrest and apoptosis in C6 rat glioma cells
Source: INTERNATIONAL JOURNAL OF ONCOLOGY, 34 (6): 1681-1690 JUN 2009 Language: English
Document Type: Article
Author Keywords: berberine; apoptosis; reactive oxygen species; mitochondria membrane potential; cytochrome c
KeyWords Plus: TOPOISOMERASE-I; CANCER-CELLS; MITOCHONDRIAL PATHWAY;
DOWN-REGULATION; LEUKEMIA-CELLS; DNA-DAMAGE; INHIBITION; CORALYNE;
INDUCTION; CLEAVAGE
Abstract: The cytotoxicity of berberine on C6 rat glioma cells indicated that berberine induced morphological changes and caused cell death through G2/M arrest and apoptosis. While undergoing apoptosis, there was a remarkable accumulation of G2/M cells with the
upregulatoin of Weel but it also inhibited cyclin B, CDK1 and Cdc25c that led to G2/M arrest.
Along with cytotoxicity in C6 cells, several apoptotic events including mitochondrial cytochrome c release, activation of caspase-9, -3 and -8 and DNA fragmentation were induced. Berberine increased the levels of GADD153 and GRP 78 in C6 cells based on the examination of Western blotting and this is a major hallmark of endoplasmic reticulum (ER) stress. We also found that berberine promoted the production of reactive oxygen species and Ca2+ in C6 cells. Western blotting assay also showed that berberine inhibited the levels of anti-apoptotic protein Bcl-2 but increased the levels of pro-apoptotic protein Bax before leading to a decrease in the levels of mitochondrial membrane potential (Delta psi(m))
followed by cytochrome c release that caused the activations of capase-9 and -3 for apoptotic occurrence. The caspase-8, -9 and -3 were activated by berberine in C6 cells based on the substrate solution (PhiPhiLux-G(1)D(1), CaspaLux 8-L1D2, CaspaLux 9-M1D2 for caspase-3, -8 and -9, respectively) and analyzed by flow cytometer and each inhibitor of caspase-8, -9 and -3 led to increase the percentage of viable C6 cells after exposure to berberine. This finding was also confirmed by Western blot assay which showed that berberine promoted the active form of caspase-8, -9 and -3. These results demonstrate that the cytotoxicity of berberine in C6 rat glioma cells is attributable to apoptosis mainly through induced G2/M- arrested cells, in an ER-dependent manner, via a mitochondria-dependent caspase pathway regulated by Bax and Bcl-2.
Addresses: [Chen, Ting-Ching; Chung, Jing-Gung] China Med Univ, Dept Biol Sci & Technol,
Taichung 404, Taiwan; [Lai, Kuang-Chi] China Med Univ, Sch Med, Taichung 404, Taiwan;
[Lai, Kuang-Chi] China Med Univ, Beigang Hosp, Dept Surg, Beigang, Yunlin, Taiwan; [Yang, Jai-Sing] China Med Univ, Dept Pharmacol, Taichung 404, Taiwan; [Liao, Ching-Lung] China Med Univ, Grad Inst Chinese Med Sci, Taichung 404, Taiwan; [Hsia, Te-Chun] China Med Univ Hosp, Dept Internal Med, Taichung, Taiwan; [Chen, Guang-Wei] Kaohsiung Med Univ, Chung Ho Mem Hosp, Dept Tradit Chinese Med, Kaohsiung, Taiwan; [Lin, Jen-Jyh] China Med Univ, Div Cardiol, Taichung, Taiwan; [Lin, Hui-Ju] China Med Univ, Dept Ophthalmol, Taichung, Taiwan; [Chiu, Tsan-Hung] China Med Univ, Dept OBS GYN, Taichung, Taiwan;
[Tang, Yih-Jing] Taichung Vet Gen Hosp, Dept Family Med, Taichung, Taiwan; [Chung, Jing- Gung] Asia Univ, Dept Biotechnol, Taichung, Taiwan
Reprint Address: Chung, JG, China Med Univ, Dept Biol Sci & Technol, 91 Hsueh Shih Rd, Taichung 404, Taiwan.
E-mail Address: [email protected] Funding Acknowledgement:
Funding Agency Grant Number
China Medical University, Taichung, Taiwan
CMU96-087 CMU97-086 CMU97-087
This study was supported by grants CMU96-087 and CMU97-086 and CMU97-087 from China Medical University, Taichung, Taiwan.
Cited References: CHEN JC, 2009, ORAL ONCOL, V45, P273, DOI 10.1016/j.oraloncology.2008.05.012.
CHUANG JY, 2005, IN VIVO, V19, P683.
CRAIG WJ, 1997, J AM DIET ASSOC S2, V97, S199.
CRAIG WJ, 1999, AM J CLIN NUTR S, V70, S491.
DAVIDSON MW, 1977, NUCLEIC ACIDS RES, V4, P2697.
FOIJER F, 2007, CANCER RES, V67, P9244, DOI 10.1158/0008-5472.CAN-07-1650.
FUKUDA K, 1999, J ETHNOPHARMACOL, V66, P227.
FUKUDA K, 1999, PLANTA MED, V65, P381.
GATTO B, 1996, CANCER RES, V56, P2795.
HUANG WM, 1989, ZHONGHUA XIN XUE GUA, V17, P300.
IHMELS H, 2005, PHOTOCHEM PHOTOBIOL, V81, P1107, DOI 10.1562/2005-01-25-IR-427.
IKEDA K, 1999, BIOCHEM PHARMACOL, V57, P1361.
KIM SA, 1998, BIOCHEMISTRY-US, V37, P16316.
KOBAYASHI Y, 1995, PLANTA MED, V61, P414.
KREY AK, 1969, SCIENCE, V166, P755.
KROEMER G, 1999, J NATL CANCER I, V91, P743.
LEE JH, 2008, ANTICANCER RES, V28, P1701.
LI TK, 2000, BIOCHEMISTRY-US, V39, P7107.
LIN CC, 2006, ANTICANCER RES, V26, P1097.
LIN CC, 2007, ANTICANCER RES, V27, P3371.
LIN JG, 1999, AM J CHINESE MED, V27, P265.
LIN JP, 2006, WORLD J GASTROENTERO, V12, P21.
LIN JP, 2008, IN VIVO, V22, P223.
LIN SS, CANC LETT IN PRESS.
LIN YT, 2007, IN VIVO, V21, P1053.
MAKHEY D, 1996, BIOORGAN MED CHEM, V4, P781.
MANTENA SK, 2006, CARCINOGENESIS, V27, P2018, DOI 10.1093/carcin/bgl043.
MULLER I, 1998, INT J MOL MED, V1, P491.
NISHINO H, 1986, ONCOLOGY, V43, P131.
PASQUAL MS, 1993, MUTAT RES, V286, P243.
PAYNE SR, 2008, MOL CELL BIOL, V28, P258, DOI 10.1128/MCB.01536-07.
SETH SD, 2004, INDIAN J MED RES, V120, P9.
TAKASE H, 1993, NIPPON YAKURIGAKU ZA, V2, P101.
TAN SL, 1998, J CELL BIOL, V141, P1423.
WANG LK, 1996, CHEM RES TOXICOL, V9, P75.
WANG SC, 2006, MUTAT RES-FUND MOL M, V593, P9, DOI 10.1016/j.mrfmmm.2005.06.023.
YU FS, 2007, IN VIVO, V21, P407.
ZHANG RX, 1990, CHINESE MED J-PEKING, V103, P658.
Cited Reference Count: 38 Times Cited: 0
Publisher: PROFESSOR D A SPANDIDOS
Publisher Address: 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE ISSN: 1019-6439
DOI: 10.3892/ijo_00000299
29-char Source Abbrev.: INT J ONCOL ISO Source Abbrev.: Int. J. Oncol.
Source Item Page Count: 10 Subject Category: Oncology ISI Document Delivery No.: 447FB
