Author(s): Sun, HL (Sun, Hui-Lung); Tsai, AC (Tsai, An-Chi); Pan, SL (Pan, Shiow-Lin); Ding, QQ (Ding, Qingqing); Yamaguchi, H (Yamaguchi, Hirohito); Lin, CN (Lin, Chun-Nan); Hung, MC (Hung, Mien-Chie); Teng, CM (Teng, Che-Ming)
Title: EPOX Inhibits Angiogenesis by Degradation of Mcl-1 through ERK Inactivation Source: CLINICAL CANCER RESEARCH, 15 (15): 4904-4914 AUG 1 2009
Language: English Document Type: Article
KeyWords Plus: ENDOTHELIAL-CELLS; DOWN-REGULATION; CANCER-CELLS;
XANTHONE DERIVATIVES; BCL-2 PROTEINS; BREAST-CANCER; TUMOR-GROWTH;
APOPTOSIS; BAX; EXPRESSION
Abstract: Purpose: Antiangiogenic therapy is considered as an effective strategy for controlling the growth and metastasis of tumors. Among a myriad of biological activities described for xanthone derivatives, the anticancer activity is quite remarkable, but the molecular mechanism is not clearly resolved. In the present study, we investigated the antiangiogenic mechanism of 3,6-di(2,3-epoxypropoxy)xanthone (EPOX), a novel Mcl-1 targeting drug.
Experimental Design: To evaluate the antiangiogenic activity of EPOX, we did cell viability, cell cycle, tube formation assay in vitro, and Matrigel plug assay in vivo. To evaluate the effect of EPOX on the endothelial signaling pathway, we did immunoblotting, immunoprecipitation, and immunofluorescence analysis. Intracellular glutathione levels were determined with the use of monochlorobimane, a glutathione-specific probe.
Results: EPOX induced endothelial cell apoptosis in association with proteasome-dependent Mcl-1 degradation. Down-regulation of Mcl-1 resulted in an increase in Mcl-1-free Bim, activation of Bax, and then signaling of mitochondria-mediated apoptosis. Additionally, glutathione depletion and extracellular signal-regulated kinase (ERK) inactivation was
observed in EPOX-treated cells. Glutathione supplementation reversed the inhibitory effects of EPOX on ERK, which increases the phosphorylation of Mcl-1 at T-163. Overexpression of mitogen-activated protein/ERK kinase (MEK) partially reversed the effect of EPOX on Mcl-1 dephosphorylation, ubiquitination, and degradation, further implicating ERK in the regulation of Mcl-1 stability.
Conclusions: This study provides evidence that EPOX induces glutathione depletion, ERK inactivation, and Mcl-1 degradation on endothelial cells, which leads to inhibition of angiogenesis. Our results suggest that EPOX is a novel antiangiogenic agent, making it a promising lead compound for further development in the treatment of angiogenesis-related pathologies.
Addresses: [Sun, Hui-Lung; Tsai, An-Chi; Pan, Shiow-Lin; Teng, Che-Ming] Natl Taiwan Univ, Coll Med, Inst Pharmacol, Taipei 100, Taiwan; [Sun, Hui-Lung; Ding, Qingqing;
Yamaguchi, Hirohito; Hung, Mien-Chie] Univ Texas MD Anderson Canc Ctr, Dept Mol &
Cellular Oncol, Houston, TX 77030 USA; [Lin, Chun-Nan] Kaohsiung Med Univ, Sch Pharm, Kaohsiung, Taiwan; [Hung, Mien-Chie] China Med Univ & Hosp, Ctr Mol Med, Taichung, Taiwan; [Hung, Mien-Chie] China Med Univ & Hosp, Grad Inst Canc Biol, Taichung, Taiwan;
[Hung, Mien-Chie] Asia Univ, Dept Biotechnol, Taichung, Taiwan
Reprint Address: Teng, CM, Natl Taiwan Univ, Coll Med, Inst Pharmacol, 1 Sect,1 Jen Ai Rd, Taipei 100, Taiwan.
E-mail Address: [email protected] Funding Acknowledgement:
Funding Agency Grant Number
National Science Council of Taiwan 96-2628-B002108-MY2 Ministry of Education Executive Yuan
Grant support: National Science Council of Taiwan grant 96-2628-B002108-MY2, and Taiwan Merit Scholarship from Ministry of Education Executive Yuan (H.-L. Sun).
Cited References: ADAMS JM, 2007, CURR OPIN IMMUNOL, V19, P488, DOI 10.1016/j.coi.2007.05.004.
BAGULEY BC, 2003, LANCET ONCOL, V4, P141.
BROWDER T, 2000, CANCER RES, V60, P1878.
BRUGAROLAS J, 2007, NEW ENGL J MED, V356, P185.
CHEN L, 2005, MOL CELL, V17, P393, DOI 10.1016/j.molcel.2004.12.030.
CHEN S, 2007, CANCER RES, V67, P782, DOI 10.1158/0008-5471CAN-06-3964.
CHEN WY, 2005, EUR J PHARMACOL, V522, P20, DOI 10.1016/j.ejphar.2005.08.047.
DIMMELER S, 2000, CIRC RES, V87, P434.
DING QQ, 2007, MOL CELL BIOL, V27, P4006, DOI 10.1128/MCB.00620-06.
DING QQ, 2008, CANCER RES, V68, P6109, DOI 10.1158/0008-5472.CAN-08-0579.
DOMINA AM, 2004, ONCOGENE, V23, P5301, DOI 10.1038/sj.onc.1207692.
DONG LF, 2007, CANCER RES, V67, P11906, DOI 10.1158/0008-5472.CAN-07-3034.
DUVAL H, 2003, ANGIOGENESIS, V6, P171.
FOLKMAN J, 2002, SEMIN ONCOL S16, V29, P15, DOI 10.1053/sonc.2002.37263.
FOLKMAN J, 2003, SEMIN CANCER BIOL, V13, P159.
GOLLOB JA, 2006, SEMIN ONCOL, V33, P392, DOI 10.1053/j.seminoncol.2006.04.002.
HAMMOND CL, 2001, J HEPATOL, V34, P946.
HUANG YT, 2005, J BIOL CHEM, V280, P2771, DOI 10.1074/jbc.M408850200.
KERBEL RS, 2004, NAT REV CANCER, V4, P423, DOI 10.1038/nrc1369.
KERBEL RS, 2008, NEW ENGL J MED, V358, P2039.
KUWANA T, 2005, MOL CELL, V17, P525, DOI 10.1016/j.molcel.205.02.003.
LI QM, 2007, J NEUROSCI, V27, P8395, DOI 10.1523/JNEUROSCI.2478-07.2007.
LIN CN, 1996, J PHARM PHARMACOL, V48, P539.
LIU HS, 1997, ANTICANCER RES, V17, P1107.
MCELIGOT AJ, 2005, ANNU REV NUTR, V25, P261, DOI 10.1146/annurev.nutr.25.050304.092633.
MCKEAGE MJ, 2008, BRIT J CANCER, V99, P2006, DOI 10.1038/sj.bjc.6604808.
MEHDI MZ, 2007, CELL BIOCHEM BIOPHYS, V47, P1.
MENG XW, 2007, J BIOL CHEM, V282, P29831, DOI 10.1074/jbc.M706110200.
MIKULA M, 2001, EMBO J, V20, P1952.
MORAN LK, 2001, CURR MED CHEM, V8, P763.
NGUYEN M, 2007, P NATL ACAD SCI USA, V104, P19512, DOI 10.1073/pnas.0709443104.
OKADA H, 2004, NAT REV CANCER, V4, P592, DOI 10.1038/nrc1412.
OSBAND AJ, 2003, SHOCK, V20, P269, DOI 10.1097/01.shk.0000079422.72656.66.
OUYANG F, 2006, CANCER RES, V66, P378, DOI 10.1158/0008-5472.CAN-05-1578.
PENG CY, 2008, CANCER LETT, V263, P114, DOI 10.1016/j.canlet.2007.12.028.
PINTO MMM, 2005, CURR MED CHEM, V12, P2517.
SHTIVELBAND MI, 2003, J THROMB HAEMOST, V1, P2225.
SUN HL, 2007, ONCOGENE, V26, P3941, DOI 10.1038/sj.onc.1210169.
VANDELFT MF, 2006, CANCER CELL, V10, P389, DOI 10.1016/j.ccr.2006.08.027.
WOLTER KG, 1997, J CELL BIOL, V139, P1281.
WU GY, 2004, J NUTR, V134, P489.
YAMAGUCHI H, 2002, CANCER RES, V62, P466.
YU CR, 2005, ONCOGENE, V24, P6861, DOI 10.1038/sj.onc.1208841.
Cited Reference Count: 43 Times Cited: 0
Publisher: AMER ASSOC CANCER RESEARCH
Publisher Address: 615 CHESTNUT ST, 17TH FLOOR, PHILADELPHIA, PA 19106-4404 USA
ISSN: 1078-0432
DOI: 10.1158/1078-0432.CCR-09-0269
29-char Source Abbrev.: CLIN CANCER RES ISO Source Abbrev.: Clin. Cancer Res.
Source Item Page Count: 11 Subject Category: Oncology ISI Document Delivery No.: 482CN
