Cinnamomum verum component 2-methoxycinnamaldehyde: a novel antiproliferative drug inducing cell death through targeting both topoisomerase I and II in human colorectal adenocarcinoma COLO 205 cells.
BACKGROUND
Cinnamomum verum is used to manufacture the spice cinnamon. In addition, the plant has been used as a Chinese herbal medication.


METHODS
We investigated the antiproliferative effect of 2-methoxycinnamaldehyde (2-MCA), a constituent of the cortex of the plant, and the molecular biomarkers associated with tumorigenesis in human colorectal adenocarcinoma COLO 205 cells. Specifically, cell viability was evaluated by colorimetric assay; apoptosis was determined by flow cytometry and morphological analysis with bright field, acridine orange, and neutral red stainings, as well as comet assay; topoisomerase I activity was determined by assay based upon DNA relaxation and topoisomerase II by DNA relaxation plus decatentation of kinetoplast DNA; lysosomal vacuolation and volume of acidic compartments (VACs) were determined by neutral red staining.


RESULTS
The results demonstrate that 2-MCA inhibited proliferation and induced apoptosis as implicated by mitochondrial membrane potential (ΔΨm) loss, activation of both caspase-3 and -9, increase of annexin V(+)PI(+) cells, as well as morphological characteristics of apoptosis. Furthermore, 2-MCA also induced lysosomal vacuolation with elevated VAC, cytotoxicity, and inhibitions of topoisomerase I as well as II activities. Additional study demonstrated the antiproliferative effect of 2-MCA found in a nude mice model.


CONCLUSIONS
Our data implicate that the antiproliferative activity of 2-MCA in vitro involved downregulation of cell growth markers, both topoisomerase I and II, and upregulation of pro-apoptotic molecules, associated with increased lysosomal vacuolation. In vivo 2-MCA reduced the tumor burden that could have significant clinical impact. Indeed, similar effects were found in other tested cell lines, including human hepatocellular carcinoma SK-Hep-1 and Hep 3B, lung adenocarcinoma A549 and squamous cell carcinoma NCI-H520, and T-lymphoblastic MOLT-3 (results not shown). Our data implicate that 2-MCA could be a potential agent for anticancer therapy.
Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.|School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan ROC.|Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan ROC.
Kuen-Daw
KD
Tsai
Faculty of Medicine, Medical University of Lublin, Lublin, Poland.
Jonathan
J
Cherng
Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.
Yi-Heng
YH
Liu
Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.
Ta-Wei
TW
Chen
Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.
Ho-Yiu
HY
Wong
Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.|School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan ROC.
Shu-Mei
SM
Yang
Department of Family Medicine, Saint Mary's Hospital Luodong, Yilan, Taiwan ROC.
Kuo-Shen
KS
Chou
Department of Internal Medicine; Saint Mary's Hospital Luodong, Yilan, Taiwan ROC; happy.professor@yahoo.com.
Jaw-Ming
JM
Cherng
2-methoxycinnamaldehyde; antiproliferative; cytotoxicity; lysosomal vacuolation; topoisomerase I; topoisomerase II; xenograft