Activation of JNK contributes to evodiamine-induced apoptosis and G2/M arrest in human colorectal carcinoma cells: a structure-activity study of evodiamine

Chih-Chiang Chien; Ming-Shun Wu; Shing-Chuan Shen; Ching-Huai Ko; Chih-Hung Chen; Ling-Ling Yang; Yen-Chou Chen

doi:10.1371/journal.pone.0099729

Activation of JNK contributes to evodiamine-induced apoptosis and G2/M arrest in human colorectal carcinoma cells: a structure-activity study of evodiamine

PLoS One. 2014 Jun 24;9(6):e99729. doi: 10.1371/journal.pone.0099729. eCollection 2014.

Authors

Chih-Chiang Chien¹, Ming-Shun Wu², Shing-Chuan Shen³, Ching-Huai Ko⁴, Chih-Hung Chen⁴, Ling-Ling Yang⁵, Yen-Chou Chen⁶

Affiliations

¹ Department of Nephrology, Chi-Mei Medical Center, Tainan, Taiwan; Department of Food Nutrition, Chung Hwa University of Medical Technology, Tainan, Taiwan.
² Division of Gastroenterology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan.
³ Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
⁴ Strategic Business and Innovation Technology Development Division, and Biomedical Technology and Device Research Labs, Industrial Technology Research Institute, Hsinchu, Taiwan.
⁵ College of LOHAS, Fo Guang University, Yilan, Taiwan.
⁶ Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan; Cancer Research Center and Orthopedics Research Center, Taipei Medical University Hospital, Taipei, Taiwan.

Abstract

Evodiamine (EVO; 8,13,13b,14-tetrahydro-14-methylindolo[2'3'-3,4]pyrido[2,1-b]quinazolin-5-[7H]-one derived from the traditional herbal medicine Evodia rutaecarpa was reported to possess anticancer activity; however, the anticancer mechanism is still unclear. In this study, we investigated the anticancer effects of EVO on human colon COLO205 and HT-29 cells and their potential mechanisms. MTT and lactate dehydrogenase (LDH) release assays showed that the viability of COLOL205 and HT-29 cells was inhibited by EVO at various concentrations in accordance with increases in the percentage of apoptotic cells and cleavage of caspase-3 and poly(ADP ribose) polymerase (PARP) proteins. Disruption of the mitochondrial membrane potential by EVO was accompanied by increased Bax, caspase-9 protein cleavage, and cytochrome (Cyt) c protein translocation in COLO205 and HT-29 cells. Application of the antioxidant N-acetyl-L-cysteine (NAC) inhibited H2O2-induced reactive oxygen species (ROS) production and apoptosis, but did not affect EVO-induced apoptosis of COLO205 or HT-29 cells. Significant increases in the G2/M ratio and cyclinB1/cdc25c protein expression by EVO were respectively identified in colon carcinoma cells via a flow cytometric analysis and Western blotting. Induction of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) protein phosphorylation was detected in EVO-treated cells, and the JNK inhibitor, SP600125, but not the ERK inhibitor, U0126, inhibited EVO-induced phosphorylated JNK protein expression, apoptosis, and G2/M arrest of colon carcinoma cells. Data of the structure-activity analysis showed that EVO-related chemicals containing an alkyl group at position 14 were able to induce apoptosis, G2/M arrest associated with increased DNA ladder formation, cleavage of caspase-3 and PARP, and elevated cycB1 and cdc25c protein expressions in COLO205 and HT-29 cells. Evidence supporting JNK activation leading to EVO-induced apoptosis and G2/M arrest in colon carcinoma cells is provided, and alkylation at position 14 of EVO is a critical substitution for treatment of colonic cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alkylation
Animals
Antineoplastic Agents, Phytogenic / pharmacology*
Apoptosis
Cell Line, Tumor
Cell Proliferation / drug effects
Colorectal Neoplasms / metabolism*
Colorectal Neoplasms / pathology
HT29 Cells
Humans
M Phase Cell Cycle Checkpoints / drug effects*
MAP Kinase Signaling System / drug effects*
Mice
NIH 3T3 Cells
Quinazolines / pharmacology*
Structure-Activity Relationship

Substances

Antineoplastic Agents, Phytogenic
Quinazolines
evodiamine

Grants and funding

This study was supported by the National Science Council of Taiwan (NSC 101-2320-B-038-025-MY3 and NSC 102-2320-B-038-050), Taipei Medical University-Wan Fang Hospital (103TMU-WFH-01-2), and Taipei Medical University-Chi Mei Hospital (103CM-TMU-04). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.