Induction of Apoptosis and Cell Cycle Arrest by <i>Jageum-Jung</i> in HepG2 Hepatoma Cells

Cho, Young-kee; Jeon, Ji-young; Shin, Yong-jeen; Seol, Jae-kyun; Rhee, Jae-hwa; Won, Jin-hee; Moon, Goo

The Journal of Internal Korean Medicine > Volume 28(4); 2007 > Article

Original Article

The Journal of Internal Korean Medicine 2007;28(4): 694-708.

紫金錠이 간암세포주 HepG2의 세포고사 및 세포주기에 미치는 영향

조영기, 전지영, 신용진, 설재균, 이재화, 원진희, 문구

원광대학교 한의과대학 내과학교실

Induction of Apoptosis and Cell Cycle Arrest by Jageum-Jung in HepG2 Hepatoma Cells

Young-kee Cho, Ji-young Jeon, Yong-jeen Shin, Jae-kyun Seol, Jae-hwa Rhee, Jin-hee Won, Goo Moon

Dept. of Internal Medicine, College of Korean Medicine, Wonkwang University

Correspondence

Goo Moon ,Tel: 063-270-1014, Fax: 063-270-1594, Email: gmoon@wonkang.ac.kr

Published online: December 30, 2007.

ABSTRACT

Objectives:
Jageum-Jung is used as an anti-cancer agent in oriental medicine, but the mechanism by which it induces cel l death in cancer cells is still unclear. The purpose of this study was to investigate the effects of Jageum-Jung on apoptosis an d cell cycle arrest in HepG2 hepatoma cells.

Methods :
Various cancer cell lines including HepG2, C6 glioma, SH-SY5Y, PANC-1, and MCF-7 cells, were used. Apopt osis was determined by DAPI nuclei staining and flow cytometry in HepG2 cells treated with various concentrations (from 25 to 200 ㎍/㎖) of H₂O extract of Jageum-Jung (JGJ) for 48 hrs. Expression of cell cycle arrest mediators including Rb, p53, p2 1, cyclin B1, cdk4, and cyclin E proteins were measured by Western blot analysis. To estimate intracellular hydrogen peroxide levels and intracellular nitric oxide levels, HepG2 cells were stained with DCFH-DA dye and DAF dye, subjected on flow cyto metric analysis.

Results:
1. Jageum-Jung decreased the viability of HepG2 cells in a dose-dependent manner. 2. Jageum-Jung induced the catalytic activation of caspase-3 in HepG2 cells. 3.. Jageum-Jung increased the intracellular hydrogen peroxide and NO in HepG2 cells. 4. Jageum-Jung increased the expression of Rb, p53 and p21 in HepG2 cells. 5. Jageum-Jung induced the expression of cyclin B1, cdk4, and cyclin E in HepG2 cells.

Conclusions:
Taken together, we suggest that Jageum-Jung exhibits cytotoxic effects on HepG2 cells, causing apoptosis and cell cycle arrest. The results showed that Jageum-Jung may do so by regulating the expression of specific target molecules that promote efficient apoptotic cell death following G₂/M phase arrest in a dose-dependent manner.

Key words: Jageum-Jung, HepG2 cells, Apoptosis, Cell Cycle Arrest, ROS