Browse > Article

Induction of Apoptosis and Its Mechanism by Siegesbeckia Glabrescens in HepG2 cells  

Kim, Yoon-Tae (Department of Physiology, College of Oriental Medicine, Dongguk University)
Lee, Heon-Jae (Department of Physiology, College of Oriental Medicine, Dongguk University)
Kim, Gil-Whon (Department of Physiology, College of Oriental Medicine, Dongguk University)
Shin, Heung-Mook (Department of Physiology, College of Oriental Medicine, Dongguk University)
Publication Information
Journal of Physiology & Pathology in Korean Medicine / v.19, no.3, 2005 , pp. 640-646 More about this Journal
Abstract
This study was performed for the investigation of anticancer effects of Siegesbeckia glabrescens(SG) on HepG2 cells, a human hepatoma cell line. In the previous study, we examined the involvement of nitric oxide (NO) on anti-proliferative and apoptotic efficacy of SG in vascular smooth muscle cells. The possible mechanism of the apoptotic effects of SG was investigated in HepG2 cells. SG showed potent cytotoxic activity in HepG2 but not chang cells, liver normal cells. SG treatment caused morphological change such as cell shrinkage, nuclei condensation and cell blebbing in HepG2 cells. SG also increased the nitrite production of HepG2 cells in a dose-dependent manner. Furthermore, L-NNA treatment inhibited the anti-proliferative effect of SG. From RT-PCR, SG decreased Bcl-2 but no affected on Bax. Western blot for procaspase-3 and COX-2 showed that degradation of procaspase-3 protein level or inhibition of COX-2 protein expression by SG treatment. In addition, the apoptotic effect of SG was also demonstrated by DNA laddering. In conclusion, SG-induced HepG2 cells death can occur via apoptosis which was dose-dependent, and associated with apoptosis-related Bcl-2/Bax gene expressions, COX-2 inhibition, caspase-3 activation and NO pathway. These results suggest that SG is potentially useful as a chemotherapeutic/chemopreventive agent in hepatocellular carcinoma.
Keywords
Siegesbeckia glabrescens; HepG2; apoptosis; Bcl-2; procaspase-3; COX-2;
Citations & Related Records
연도 인용수 순위
  • Reference
1 박래길, 오광록, 이광규, 문연자, 김정훈, 우원흥, 유향 물추출물의 HL-60 혈액암세포에서 세포사멸 유도효과, 약학회지, 45(2):161-168, 2001.
2 Devitt, G.P., Creagh, E.M., Cotter, T.G. Antioxidant 4b,5,9b,10 -tetrahydroindenol{1,2-b}indole inhibits apoptosis by preventing caspase activation following mitochondrial depolarization. Biochemical and Biophysical Communications, 264, 622-629, 1999.
3 김호철, 생희첨과 주증희첨 추출물이 자발성 고혈압 흰쥐의 항 고혈압 작용에 미치는 영향, 본초학회지 13(1):53-58, 1998.
4 김호현, 김길훤, 희렴이 가토의 혈관내피세포성 이완인자에 미치는 영향, 대한한의학회지,18(2):15-32, 1997.
5 신동훈, 희렴의 Nitric Oxide 유리를 통한 평활근세포에서의 Apoptosis 유도, 동국대학교 대학원,2003.
6 Semon, Wu, Yu-Shien, Ko, Ming-Sheng, Teng, Yu-Lin, Ko, Lung-An, Hsu, Chueh, Yung-Yin. Chou, Choong-Chin. Liew and Ying-Shiung, Lee. Adriamycin-induced Cardiomyocyte and Endothelial Cell Apoptosis: In vitro and In vivo studies. J Mol Cell cardiol. 34, 1595-1607, 2002.
7 郁仁存. 中醫腫瘤學, 上海, 상해과학기술출판사, pp 1-22, 41-44, 384-391, 1991.
8 허연진, 김정환, 장정희, 안병준, 서영준. 새로운 Anthraquinone 유도체, DHAQ-97의 항암작용: 아폽토시스에 의한 인체 유방암 세표 사멸 유도 Environmental Mutagens &Carcinogens 20(1) :14-20, 2000.
9 Chiarugi, V., magneil, L., Cinelli, Basi G. Apoposis and the cell cycle. Cell. Mol. Biol. Res. 40:603-612, 1994.
10 張民厭, 龔惠明 主編. 抗腫瘤中藥的臨床應用, 北京, 人民衛生出版社, 1998.
11 Esperanza, R. Matarredona, Maribel Murilo-Carretero, Bernardo Moreno-Lopez, Carmen Estrada. Nitric Oxide synthesis inhibition increase proliferation of neural precursors isolated from the postnatal mouse subventricular zone. Brain Research, 995, 274-284, 2004.
12 Thompson, C.B. Apoptosis in the pathogenesis and treatment of dosage. Science, 267, 1456-1462, 1995.
13 Chen, Q., Yang, G.W., An, L.G. Apoptosis of hepatoma cells SMMC-7721 induced by Ginkgo biloba seed polysaccharide. World J Gastroenterol, 8, 832-836, 2002.
14 Griess, P. Bemerkungen zu der abhandlung der H.H. Weselsky und Benedikt. 'Ueber einige azoverbindungen.' Chem. Ber. 12, 426. 1879.
15 Taketo, M.M. Cyclooyxgenase-2 inhibition in tumorigenesis (part II), J Natl Cancer Inst, 90, 1609-1620, 1998.
16 Wanebo, H.J., Falkson, S.E. Cancer: Principles and Practice of Oncology, vol.1. Lippincott, London, pp 836-870, 1989.
17 Antonsson, B., Martinou, J.C. The Bcl-2 protein family. Exp. Cell Res, 256, 50-57, 2000.
18 Qui, D.K., Ma, X., Peng, Y.S. Chen, X.Y. Significance of cyclooxygenase-2 expression in human primay hepatocellular carcinoma. World J Gastroenterol, 8, 815-817, 2002.
19 Kurose, I., Higuchi, H., et al. Rat Kupffer cell-derived nitric oxide suppresses proliferation and induces apoptosis of syngeneic hepatoma cells. Gastroenterology. 111(4):1058-1070, 1996.
20 저수금, 惡性腫瘤中醫調理四法, 상해중의약잡지 7, 33-34, 1992.
21 Lowery, O.H. et al. Protein measurement with the Folin phenol reagent. J BioI Chem. 193, 265-275. 1951.
22 Planchon, S.M., Wuerzberger, S., Frydman, B., Witiak, D.T., Hutson, P., Church, Dr, Wilding, G., Boothman, D.A. $\beta$ -Lapachone-mediated apoptosis in human promyelocytic leukemia(HL-60) and human prostate cancer cells: p-53-independent response. Cancer Res., 55, 3706-3711, 1995.
23 高學敏. 中藥學, 北京, 北京中醫學院, pp 155-156, 1986.
24 金臭顯, 申興默, 金吉萱. 稀簽이 收縮된 血管에 미치는 影響, 東醫生理學會誌, 10(1):25-36, 1995.
25 劉嘉湘 主編. 實用中醫學腫瘤手冊, 上海, 上海科技教育出版社, pp 1-7, 1996.
26 신흥묵. 豨簽의 혈관이완 효능과 항산화 동태에 관한 연구, 대한한의학회지 21(1):77-83, 2000.
27 Taketo, M.M. Cyclooyxgenase-2 inhibition in tumorigenesis (part I), J Natl Cancer Inst, 90, 1529-1536, 1998.
28 Koga, H., Sakisaka, S., Ohishi, M., Kawaguchi, T., Taniguchi, E., Sasatomi, K., Harada, M., Kusabab, T., Tanaka, M., Kimura, R, Nakasima, Y., Nakasima, O., Kojiro. M., Kurohiji, T., Sata, M. Expression of cyclooxygenase-2 in human hepatocellular carcinoma: relevance to tumar defifferentiation, Hepatology 29, 688-696, 1999.
29 남상수, 박동석, 희렴약침 자극이 자발성 고혈압 흰쥐의 혈압에 미치는 영향 및 강압기전에 대한 연구, 대한한의학회지, 18(1):218-237, 1997.
30 Muller, I., Jenner, A., Bruchelt, G., Niethammer, D., Halliwell, B. Effect of concentration on the cytotoxic mechanism of doxorubicin-apoptosis and oxidative DNA damage. Biochem. Biophy. Res. Commun. 230, 254-257, 1977.
31 趟健娥. 吳一純敎授治療晚期惡性腫瘤的經驗, 陝西醫學:,14, 451-453, 1993.
32 Scudiero, D. A., Shoemaker, R. H., Paull, K. D., Monks, A., Tierney, S., Nofziger, T. H., Currens, M. J., Seniff, D., Boyd, M. R. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Research. 48, 4827-4833. 1988.
33 Herrmann, M., Lorenz, H.M., Vollm, R., Grunkem, M., Woithm, W., Kaldenm, J.R. A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res. 22, 5506-5507, 1994.
34 GeneBank Accesion No: M13994, Human Bcl-2 complete CDS.