Arctigenin Anti-Tumor Activity in
Bladder Cancer T24 Cell Line Through
Induction of Cell-Cycle Arrest and
SHUCAI YANG,1JING MA,1JIANBING XIAO,1XIAOHONG LV,1XINLEI LI,1
HUIKE YANG,1YING LIU,2SIJIA FENG,1AND YAFANG ZHANG1*
1Department of Anatomy, Harbin Medical University, Harbin, China
2Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical
University, Harbin, China
Bladder cancer is the most common neoplasm in the urinary system.
This study assesses arctigenin anti-tumor activity in human bladder can-
cer T24 cells in vitro and the underlying molecular events. The flow cytom-
etry analysis was used to detect cell-cycle distribution and apoptosis.
Western blotting was used to detect changes in protein expression. The
data showed that arctigenin treatment reduced viability of bladder cancer
T24 cells in a dose- and time-dependent manner after treatment with arcti-
genin (10, 20, 40, 80, and 100 lmol/L) for 24 hr and 48 hr. Arctigenin treat-
ment clearly arrested tumor cells in the G1 phase of the cell cycle.
Apoptosis was detected by hoechst stain and flow cytometry after Annexin-
V-FITC/PI double staining. Early and late apoptotic cells were accounted
for 2.32–7.01% and 3.07–7.35%, respectively. At the molecular level, arcti-
genin treatment decreased cyclin D1 expression, whereas CDK4 and CDK6
expression levels were unaffected. Moreover, arctigenin selectively altered
the phosphorylation of members of the MAPK superfamily, decreasing
phosphorylation of ERK1/2 and activated phosphorylation of p38 signifi-
cantly in a dose-dependent manner. These results suggest that arctigenin
may inhibit cell viability and induce apoptosis by direct activation of the
mitochondrial pathway, and the mitogen-activated protein kinase pathway
may play an important role in the anti-tumor effect of arctigenin. The data
from the current study demonstrate the usefulness of arctigenin in bladder
cancer T24 cells, which should further be evaluated in vivo before transla-
tion into clinical trials for the chemoprevention of bladder cancer. Anat
Rec, 295:1260–1266, 2012.
C2012 Wiley Periodicals, Inc.
Key words: arctigenin; cyclinD1; ERK1/2; p38; bladder cancer
Bladder cancer is the most common neoplasm of the
urinary system and also a significant health problem.
Histologically, 95% of bladder cancers are transitional
cell carcinomas (Jernal et al., 2004). Tobacco smoking is
the main known contributor to bladder cancer, and the
occupations at risk are bus driver, rubber worker, motor
mechanics, leather worker, or blacksmith. Citrus fruits
and yellow–orange vegetables, particularly carrots and
selenium have a protective effect for bladder cancer.
Treatment for bladder cancer depends on the stage of
the disease and tumor characteristics, but bladder can-
cer frequently recurs after surgery. Bacillus Calmette
Guerin vaccine (BCG) immunotherapy is effective in up
to two thirds of bladder cancer cases, and a combination
*Correspondence to: Yafang Zhang, 194 Xuefu Road, Harbin,
China. E-mail: email@example.com
Received 15 January 2012; Accepted 16 April 2012.
Published online 23 May 2012 in Wiley Online Library
THE ANATOMICAL RECORD 295:1260–1266 (2012)
C 2012 WILEY PERIODICALS, INC.
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