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

Kuen-Daw Tsai; Jonathan Cherng; Yi-Heng Liu; Ta-Wei Chen; Ho-Yiu Wong; Shu-Mei Yang; Kuo-Shen Chou; Jaw-Ming Cherng

doi:10.3402/fnr.v60.31607

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

Food Nutr Res. 2016 Jun 7:60:31607. doi: 10.3402/fnr.v60.31607. eCollection 2016.

Authors

Kuen-Daw Tsai^{1

2

3}, Jonathan Cherng⁴, Yi-Heng Liu¹, Ta-Wei Chen¹, Ho-Yiu Wong¹, Shu-Mei Yang^{1

2}, Kuo-Shen Chou⁵, Jaw-Ming Cherng⁶

Affiliations

¹ Department of Internal Medicine, China Medical University Beigang Hospital, Yunlin, Taiwan ROC.
² School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan ROC.
³ Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan ROC.
⁴ Faculty of Medicine, Medical University of Lublin, Lublin, Poland.
⁵ Department of Family Medicine, Saint Mary's Hospital Luodong, Yilan, Taiwan ROC.
⁶ Department of Internal Medicine; Saint Mary's Hospital Luodong, Yilan, Taiwan ROC; happy.professor@yahoo.com.

Abstract

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.

Keywords: 2-methoxycinnamaldehyde; antiproliferative; cytotoxicity; lysosomal vacuolation; topoisomerase I; topoisomerase II; xenograft.