Browse > Article

Effects of Rhus verniciflua Stokes Extract on Cell Viability, Cell Cycle Progression and Apoptosis of AGS Cell  

An, Jin-Yeong (Department of Internal Medicine, College of Oriental Medicine, Semyung University)
Ko, Seong-Gyu (Department of Preventive Medicine, College of Oriental Medicine, Kyunghee University)
Ko, Heung (Department of Internal Medicine, College of Oriental Medicine, Semyung University)
Publication Information
Journal of Physiology & Pathology in Korean Medicine / v.20, no.3, 2006 , pp. 701-709 More about this Journal
Abstract
The Rhus verniciflua Stokes (乾漆-RVS) has been used in traditional East Asia medicine for the therapy of gastritis, stomach cancer, although the mechanism for the biological activity is unclear. In the present study aims to investigate RVS extract contributes to growth inhibitory effect and it's the molecular mechanism on the human gastric cancer cells. AGS (gastric cancer cells) and RIEI (normal cells) were treated to different concentrations and periods of RVS extract $(10{\;}{\sim{{\;}100{\;}ug/mil)$. Growth inhibitory effect was analyzed by measuring FACS study and MTS assay. Cell cycle inhibition was confirmed by measuring CDK2 kinase activity by immunoprecipitation and kinase assay. And apoptosis was confirmed by surveying caspase cascades activation using a pan caspase inhibitor Exposure to RVS extract (50 ug/mll) resulted in a synergistic inhibitory effect on cell growth in AGS cells. Growth inhibition was related with the inhibition of proliferation and induction of apoptosis. The extract induces Gl -cell cycle arrest through the regulation of cyclins, the induction of p27kip1, and the decrease CDK2 kinase activity. And upregulated p27kip1 level is caused by protein stability increment by the reduction of S-phase kinase-associated protein 2 (Skp2), a key molecule related with p27kip1 ubiquitination and degradation, and do novo protein synthesis. Besides, 乾漆 extract induces apoptosis through the expression of Bax, poly(ADP-ribose) polymerase (PARP) and activation of caspase-3. RVS extract induces Gl -cell cycle arrest via accumulation of p27kip1 and apoptosis in human gastric cancer cells but not in normal cells, therefore we suggest that the extract can be used as a novel class of anti-cancer drugs.
Keywords
Rhus verniciflua Stokes; RVS; apoptosis; cell cycle; Gl arrest; p27kip;
Citations & Related Records
연도 인용수 순위
  • Reference
1 김성훈. 韓醫學界의 癌硏究動向과 硏究戰略에 대한 연구. 대한한의학회지. 19(1):470-499, 1998
2 Sherr, C.J. and Roberts, J.M. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev.. 12:1501-1121, 1999.
3 Pagano, M., Tam, S.W., Theodoras, A.M. et al. Role of the ubiquitin-proteasome pathway in regulating abundance of the cyclin-dependent kinase inhibitor p27. Science (Wash. DC). 269:682-685, 1995   DOI
4 Sutterluty, H., Chatelain, E., Marti, A., Wirbelauer, C., Senften, M., Muller, U. and Krek, W. p45SKP2 promotes p27Kip1 degradation and induces S phase in quiescent cells. Nat. Cell Biol. 1:207-214, 1999   DOI   ScienceOn
5 Nevins, J.R. E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. Science. 258:424-429, 1992   DOI
6 Nakayama, K.I., Hatakeyama, S. and Nakayama, K. Regulation of the cell cycle at the G1-S transition by proteolysis of cyclin E and p27Kip1. Biochem Biophys Res Commun. 4:853-860, 2001
7 Vlach, J., Hennecke, S., and Amati, B. Phosphorylation- dependent degradation of the cyclin-dependent kinase inhibitor p27Kip1. EMBO J., 16:5334-5344, 1997   DOI   ScienceOn
8 Lloyd, R.V., Erickson, L.A., Jin, L. et al. p27kip1: a multifunctional cyclin dependent kinase inhibitor with prognostic significance in human cancers. Am J Pathol., 2:313-323, 1999
9 Lee, J.C., Lee, K.Y., Kim, J.N., et al. Extract from Rhus verniciflua Stokes is capable of inhibiting the growth of human lymphoma cells. Food Chem Toxicol. 9:1383-1388, 2004
10 Schulman, B.A., Carrano, A.C., Jeffrey, P.D. et al. Insights into SCF ubiquitin ligases from the structure of the Skp1-Skp2 complex. Nature (Lond.). 408:381-386, 2000   DOI   ScienceOn
11 Meier, P., Finch, A. and Evan, G. Apoptosis in development. Nature. 407:796-801, 2000   DOI   ScienceOn
12 Kitts, D.D., and Lim, K. TAntitumorigenic and cytotoxic properties of an ethanol extract derived from Rhus verniciflua Stokes (乾漆). Journal of Toxicology and Environmental Health Part A. 64:357-371, 2001   DOI   ScienceOn
13 Coats, S., Flanagen, M., Nourse, J., and Roberts, J.M. Requirement of p27kip1 for restriction point control of the fibroblast cell cycle. Science. 272:877-880, 1996   DOI
14 Polyak, K., Kato, J.Y., Solomon, M.J., Sherr, C.J., Massague, J., Roberts, J.M., and Koff, A. P27 KIP1, a cyclin Cdk inhibitor, links transforming growth factor beta and contact inhibition to cell cycle arrest. Genes Dev. 8:9-22, 1994   DOI   ScienceOn
15 Helin, K., Harlow, E. and Fattaey, A.R. Inhibition of E2F-1 transactivation by direct binding of the retinoblastoma protein. Mol. Cell. Biol. 13:6501-6508, 1993   DOI
16 Jung, N.C. Biological activity of urushiol and flavonoids from Lac tree (Rhus verniciflua Stokes). Ph.D. Thesis, Chonnam National University, Kwang-ju, South Korea, 1998
17 Lee, J.C., Kim, J. and Jang, Y.S. Ethanol-eluted extract of Rhus verniciflua stokes inhibits cell growth and induces apoptosis in human lymphoma cells. J Biochem Mol Biol.. 4:337-343, 2003
18 Toyoshima, H. and Hunter, T. p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21.Cell. 78:67-74, 1994   DOI   ScienceOn
19 김 병, 문 구. 위암의 동서의학적 진치개황. 대한한의학회지. 17(2):100-116, 1996
20 통계청. 2004년도 사망원인통계결과, 서울. 통계청, pp 13-15, 2005
21 Kothakota, S., Azuma, T., Reinhard, C. et al. Caspase- 3-generated fragment of gelsolin: Effector of morphological change in apoptosis. Science. 278:294-298, 1997   DOI
22 Coqueret, O. New roles for p21 and p27 cell-cycle inhibitors: a function for each cell compartment Trends. Cell Biol. 2:65-70, 2003
23 Tsvetkov, L.M., Yeh, K.H., Lee, S.J., Sun, H. and Zhang, H. p27(Kip1) ubiquitination and degradation is regulated by the SCF(Skp2) complex through phosphorylated Thr187 in p27. Curr. Biol. 9:661-664, 1999   DOI   ScienceOn
24 Stacy, W.B. p27 as a target for cancer therapeutics. Cancer cell. 3:111-115, 2003   DOI   ScienceOn
25 Jonstone, R.W. Apoptosis: a link between cancer genetics and chemotherapy. Cell. 108:153-164, 2002   DOI   ScienceOn
26 Atan, G., James, M.M. and Stanley, J.K. Bcl-2 family members and the mitochondria in apoptosis. Genes. Dev. 13:1899-1911, 1999   DOI   ScienceOn
27 Green, D.R. Apoptotic pathways: paper wraps stone blunts scissors. Cell. 102:1-4, 2000   DOI   ScienceOn
28 Nunez, G., Benedict, M.A., Hu, Y. and Inohara, N. Caspases: The proteases of the apoptotic pathway. Oncogene. 17:3237-3245, 1998   DOI   ScienceOn
29 윤성우, 유봉하, 박동원, 유기원. 위암이 동의학 및 동서의 결합치료에 관한 문헌적 고찰. 대한종양학회지. 2(1):177-192, 1996
30 Humbert, P.O., Verona, R., Trimarchi, J.M., Rogers, C., Dandapani, S. and Lees, J.A. E2F3 is critical for normal cellular proliferation. Genes Dev. 14:690-703, 2000
31 Lee, J.C., Kim, J., Lim, K.T., Yang, M.S. and Jang, Y.S. Ethanol eluted extract of Rhus verniciflua Stokes showed both antioxidant and cytotoxic effects on mouse thymocytes depending on the dose and time of the treatment. Journal of Biochemistry and Molecular Biology. 34:250-258, 2001
32 Elledge, S.J. and Harper, J.W. The role of protein stability in the cell cycle and cancer. Biochim. Biophys. Acta. 1377:M61-M70, 1998
33 Lee, J.C., Lim, K.T. and Jang, Y.S. Identification of Rhus verniciflua Stokes compounds that exhibit free radical scavenging and anti-apoptotic properties. Biochimica et Biophysica Acta., 1570:181-191, 2002   DOI
34 Dalton, S. Cell cycle regulation of the human cdc2 gene. EMBO J. 11:1797-1804, 1992
35 Sheaff, R.J., Groudine, M., Gordon, M., Roberts, J.M. and Clurman, B.E. Cyclin E-CDK2 is a regulator of p27Kip1. Genes Dev.. 11:1464-1478, 1997   DOI   ScienceOn
36 안덕균. 한국본초도감. 서울. 교학사, p 729, 2003
37 Nasmyth, K. Viewpoint: putting the cell cycle in order. Science. 274:1643-1645, 1996   DOI   ScienceOn
38 Masuda, T.A., Inoue, H., Sonoda, H. et al. Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma: modulation of malignant phenotype by Skp2 overexpression, possibly via p27 proteolysis. Cancer Res. 13:3819-3825, 2002
39 Flemington, E.K., Speck, S.H., and Kaelin, Jr. W.G. E2F-1-mediated transactivation is inhibited by complex formation with the retinoblastoma susceptibility gene product. Proc. Natl. Acad. Sci. USA, 90:6914-6918, 1993
40 김동희, 김성훈. 抗癌劑 및 放射線 副作用에 대한 韓方療法. 동의병리학회지. 9:239-264, 1994
41 Adams, J.M. and Cory, S. The Bcl-2 protein family: arbiters of cell survival. Science. 281:1322-1326, 1998   DOI   ScienceOn
42 Masuda, T.A., Inoue, H., Sonoda, H. et al. Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma: modulation of malignant phenotype by Skp2 overexpression, possibly via p27 proteolysis. Cancer Res. 13:3819-3825, 2002
43 Carrano, A.C., Eytan, E., Hershko, A. and Pagano, M. SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nat. Cell Biol. 1:193-199, 1999.   DOI   ScienceOn