Saussurea lappa induces G2-growth arrest and apoptosis in AGS gastric cancer cells

doi:10.1016/j.canlet.2004.06.026

Cancer Letters

Volume 220, Issue 1, 18 March 2005, Pages 11-19

https://doi.org/10.1016/j.canlet.2004.06.026 Get rights and content

Absract

The molecular effects of Saussurea lappa extracts, a traditional medicine in Eastern Asia, on the fate of gastric carcinoma have not been understood. In this study, its cytostatic effects were examined using gastric AGS cancer cells. Its treatment resulted in apoptosis and G2-arrest in a dose- and time-dependent manner. The effects were attributed to the regulation of cyclins and pro-apoptotic molecules and suppression of anti-apoptotic molecules. Therefore, these results suggest that extracts of S. lappa root may be a candidate to deal with gastric cancers either by traditional herbal therapy or by combinational therapy with conventional chemotherapy.

Introduction

The dried root of Saussurea lappa Clarke has been traditionally used for abdominal pain and tenesmus as a traditional medicine in Korea, China, and Japan. Sesquiterpene lactones including costunolide and dehydrocostus lactone are major components of the root and have been suggested to possess various biological activities, including anti-tumor, anti-ulcer, anti-inflammatory, neurocytotoxic and cardiotonic activities [1]. Costunolide has been shown to have preventive effects on intestinal carcinogenesis, via pro-apoptotic effects of costunolide [2], [3]. In addition, it has also been shown to inhibit endothelial cell proliferation via a blocking of VEGFR KDR/Flk-1 signal pathway [4].

Cell proliferation is a tightly controlled process consisting of multiple checkpoints responsible for the regulation of abnormal cell cycle progression. Transitions between G1, S, and G2/M phases are regulated by biochemically coordinated actions of cyclins, cyclin-dependent kinases (CDKs), CDK inhibitors (CKIs), all of which can in turn be modulated by diverse intracellular signals transduced from extracellular growth cues [5]. So far, the G1 to S phase transition through the restriction (R) point or S phase entry has been shown to be regulated by mitogenic reagents, an intact cytoskeletal network, and cell adhesion as well [6], [7], [8], [9], [10]. In addition to the G1-checkpoint, there has been evidence that cellular systems often arrest at a G2-check point to avoid cell divisions that would produce daughters with damaged or abnormally synthesized DNA [11], [12], [13]. One of the molecules involved in the cell growth checkpoint includes p53, which induces cell cycle regulators including p21^Waf1 CKI [13]. In turn, p21^Waf1 CKI inhibits CDC2-cylinB complex, leading to a G2-arrest [12].

On the other hand, although gastric cancer occurs very frequently and S. lappa extracts have been used as a traditional medicine in Asia, the molecular effects of S. lappa extracts on cell proliferation or apoptosis have not been precisely and fully studied. Knowledge with regards to gastric cancer and medicinal herbs can probably have an ever greater impact on clinical management. In this study, we have tried to explore how S. lappa extract influences the growth of AGS gastric cancer cells at the molecular level. We found that treatment with S. lappa extract on AGS gastric cancer cells resulted in G2-growth arrest, presumably involving p53 and p21^Waf1 CKI induction and concomitant reduction of cyclin B1. Furthermore, the extracts induced apoptosis via activation of pro-apoptotic molecules including Bax and caspase3, and suppression of anti-apoptotic Bcl₂.

Section snippets

Preparation of Saussurea lappa extracts

The extract of S. lappa root (BoKwang Crude Drugs Co., Seoul, Korea) was prepared by sonication in 80% ethanol and then a freeze-drying process. The HPLC analysis of the extracts was performed using a standard material of costunolide (Wako Pure Chemicals Ind. Co., Japan), with Breeze™ HPLC systems with a Waters 2487 Dual Wavelength Absorbance Detector (Waters Corp., Milford, MA). Commercial pure costunolide (0.5 mg/ml of acetone) or test sample of the extracts (10.0 mg/ml of acetone) were used in

Results

We have had an interest in examining the anticancer activity of S. lappa extract on a gastric cancer cell line, AGS cells, with a special emphasis on the effects of the extract on apoptosis and cell cycle arrest pathway.

We first checked whether the S. lappa extracts included Sesquiterpene lactones including costunolide by a HPLC analysis. As shown in Fig. 1A, the extracts showed major peaks including one for costunolide. Then we determined how S. lappa extract affected the viability of AGS

Discussion

We observed that treatment of S. lappa extract induced G2-arrest and apoptosis of AGS gastric cancer cells, probably by both ATM/ATR-dependent and independent pathways involving induction of p53/p21^Waf1 CKI, induction and activation of pro-apoptotic molecules, and concomitantly suppression of anti-apoptotic molecules. Meanwhile, the extract caused much less significant cytostatic effects on normal rat intestinal epithelial RIE1 cells, indicating tumor cell specific effects. These results may

Acknowledgements

This study was supported in part by a fund from Ministry of Health and Welfare, Korea (to S.G. Ko) and in part by 2003 BK21 Project for Medicine, Dentistry, and Pharmacy (to J.W. Lee).

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View full text

Saussurea lappa induces G2-growth arrest and apoptosis in AGS gastric cancer cells

Absract

Introduction

Section snippets

Preparation of Saussurea lappa extracts

Results

Discussion

Acknowledgements

Cancer Lett.

Cancer Lett.

Curr. Opin. Cell Biol.

Curr. Opin. Genet. Dev.

Exp. Cell Res.

J. Biol. Chem.

Toxicology

Recent studies on the zoopharmacognosy, pharmacology and neurotoxicology of sesquiterpene lactones

Planta Med.

Costunolide induces apoptosis by ROS-mediated mitochondrial permeability transition and cytochrome C release

Biol. Pharm. Bull.

Regulation of cyclin-Cdk activity in mammalian cells

Cell Mol. Life Sci.