[논문]Pseudolaric Acid B Induces Apoptosis Through p53 and Bax/Bcl-2 Pathways in Human Melanoma A375-S2 Cells

Gong Xian-Feng; Wang Min-Wei; Tashiro Shin-Ichi; Onodera Satoshi; Ikejima Takashi

doi:10.1007/bf02975138

Pseudolaric Acid B Induces Apoptosis Through p53 and Bax/Bcl-2 Pathways in Human Melanoma A375-S2 Cells 원문보기

Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea, v.28 no.1, 2005년, pp.68 - 72

Gong Xian-Feng (China-Japan Research Institute of Medical and Pharmaceutical Sciences, Department of Pharmacology, Shen-yang Pharmaceutical University) , Wang Min-Wei (Department of Pharmacology, Shenyang Pharmaceutical university) , Tashiro Shin-Ichi (Department of Clinical and Biomedical Sciences, Showa Pharmaceutical university) , Onodera Satoshi (Department of Clinical and Biomedical Sciences, Showa Pharmaceutical university) , Ikejima Takashi (China-Japan Research Institute of Medical and Pharmaceutical Sciences, Shen-yang Pharmaceutical University)

Abstract ▼ AI-Helper

Pseudolaric acid B is a major compound found in the bark of Pseudolarix kaempferi Gordon. In our study, pseudolaric acid B inhibited growth of human melanoma cells, A375-S2 in a time and dose-dependent manner. A375-S2 cells treated with pseudolaric acid B showed typical characteristics of apoptosis including morphologic changes, DNA fragmentation, sub-diploid peak in flow cytometry, cleavage of poly-ADP ribose polymerase (PARP) and degradation of inhibitor of caspase-activated DNase (ICAD). P53 protein expression was upregulated while cells were arrested at the $G_2/M$ phase of the cell cycle. There was a decrease in the expression of anti-apoptotic Bcl-2 and Bcl-xL proteins, whereas pro-apoptotic Bax was increased. The two classical caspase substrates, PARP and ICAD, were both decreased in a time-dependent manner, indicating the activation of downstream caspases.

주제어

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가설 설정

In this study, we investigated the apoptotic effect of ps이jd이aric acid B on human melanoma A375-S2 cells.

제안 방법

To investigate further features of cell growth inhibition by pseudolaric acid B, flow cytometric analysis was performed. After treatment with increasing concentrations of pseudolaric acid B for 24 h, the cells had accumulated in the GJM phase.

대상 데이터

Pseudolaric acid B was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Hoechst 33258, RNase A, Proteinase K, and 3-(4, 5u加1枪甘»1巾谊2이-2刑)・2, 5- diphenyltetrazolium bromide (MTT) were purchased from Sigma Chemical (St. Louis, MO). Rabbit p이yclonal antibodies against poly-ADP ribose polymeras으 (PARP) and inhibitor of caspase- activated DNase (ICAD), mouse monoclonal antibodies against Bcl-2, Bcl-xL, and p53, as well as horseradish peroxidase-conjugated secondary antibodies (goat-anti-rabbit and goat-anti-mouse) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).
Louis, MO). Rabbit p이yclonal antibodies against poly-ADP ribose polymeras으 (PARP) and inhibitor of caspase- activated DNase (ICAD), mouse monoclonal antibodies against Bcl-2, Bcl-xL, and p53, as well as horseradish peroxidase-conjugated secondary antibodies (goat-anti-rabbit and goat-anti-mouse) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit polyclonal antibody against Bax was obtained from Oncogene Research Products (Cambridge, MA).

이론/모형

The cells (5x 104 cells per well) were incubated with various concentrations of pseudolaric acid B for 12 h (♦), 24 h (■), 36 h (▲), or 48 h (□). Growth inhibition was evaluated by the MTT method.

성능/효과

Since PARP and ICAD were classical substrates of caspases, we measured the expression of PARP and ICAD by Western blot analysis. The results showed that both PARP and ICAD expression had decreased,suggesting that caspases participated in pseudolaric acid B induced A375-S2 cell death (Fig. 8).

참고문헌 (21)

Agarwal, N. and Mehta, K., Possible involvement of Bcl-2 pathway in retinoid X receptor alpha-induced apoptosis of HL-60 cells. Biochem. Biophys. Res. Comm., 230, 251-253 (1997)

상세보기
Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A., and Nagata, S., A., Caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature, 391, 43-50 (1998)

상세보기
Hermeking, H., Lengauer, C., Polyak, K., He, T. C., Zhang, L., Thiagalingam, S., Kinzler, K. W., and Vogelstein, B., 14-3-3- Sigma is a p53-regulated inhibitor of $G_2$ /M progression. Mol. Cell, 1, 3-11 (1997)

상세보기
Hockenbery, D. M., Oltvai, Z. N., Yin, X. M., Milliman, C. L., and Korsmeyer, S. J., Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell, 75, 241-251 (1993)

상세보기
Kastan, M.B., Onyekwere, O., Sidransky, D., Vogelstein, B., and Craig, R.W., Participation of p53 protein in cellular response to DNA damage. Cancer Res., 51, 6304-6311 (1991)
Kerr, J.F., Wyllie, A.H., and Currie, A.R., Apoptosis: a basic biologic phenomenon with wide-ranging implications in tissue kinetics. Br. J. Cancer, 26, 239-257 (1972)

상세보기
Lane, D.P., p53, guardian of the genome. Nature, 258, 15-16 (1992)

상세보기
Lazebnik, Y. A., Kaufmann, S. H., Desnoyers, S., Poirier, G. G., and Earnshaw, W. C., Cleavage of poly (ADP-ribose) polymerase by a proteinase with properties like ICE. Nature, 371, 346-347 (1994)

상세보기
Li, E., Clark, A. M., and Hufford, C. D., Antifungal evaluation of pseudolaric acid B, a major constituent of pseudolarix kaempferi. J. Nat. Prod., 58, 57-67 (1995)

상세보기
Liu, X. S., Zou, H., Slaughter, C., and Wang, X., DFF, a heterodimeric protein that functions downstream of caspase- 3 to trigger DNA fragmentation during apoptosis. Cell, 89, 175-184 (1997)

상세보기
Mengubas, K., Riordan, F. A., Hoffbrand, V., and Wickremasinghe, G., Co-ordinated downregulation of Bcl-2 and Bax expression during granulocytic and macrophage-like differentiation of the HL60 premyelocytic leukemia cell line. FEBS. Lett., 394, 356-360 (1996)

상세보기
Nicholson, D. W., Ali, A., Thornberry, N. A., Vaillancourt, J. P., Ding, C. K., Gallant, M., Gareau, Y., Griffin, P. R., Labelle, M., and Lazebnik, Y. A., Identification and inhibition of the ICE/ CED-3 protease necessary for mammalian apoptosis. Nature, 376, 37-43 (1995)

상세보기
Pan, D. J., Li, Z. L., Hu, C. Q., Chen, K., Chang, J. J., and Lee, K. H., The cytotoxic principles of Pseudolarix Kaempferi: Pseudolaric acid-A and B and related derivatives. Planta Med., 56, 383-385 (1990)

상세보기
Sakahira, H., Enari, M., and Nagata, S., Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature, 391, 96-99 (1998)

상세보기
Schwartz, D., Almong, N., Peled, A., Goldfinger, N., and Rotter, V., Role of wild-type p53 in the $G_{2}$ phase: regulation of the ${\alpha}$ - irradiation induced delay and DNA repair. Oncogene, 15, 2597-2607 (1997)

상세보기
Stewart, N., Hicks, G. G., Paraskevans, F., and Mowat, M., Evidence for a second cell cycle block at $G_2$ /M by p53. Oncogene, 10, 109-115 (1995)

상세보기
Taylor, W.R., and Stark, G.R., Regulation of the $G_2$ /M transition by p53. Oncogene, 20, 1803-1815 (2001)

상세보기
Wang, W. C., Lu, R.F., Zhao, S. X., and Gu, Z. P., Comparison of early pregnancy-terminating effect and toxicity between pseudolaric acids A and B. Acta. Pharmacol. Sin., 9, 445-448 (1988)
Wang, W. C., Lu, R. F., Zhao, S. X., and Zhu, Y. Z., Antifertility effect of pseudolaric acid B. Acta. Pharmacol. Sin., 3, 188- 192 (1982)
Wu, L. X., Xu, J. H., Wu, G. H., and Chen, Y. Z., Inhibitory effect of curcumin on proliferation of K562 cells involves downregulation of $p210^{bcr/abl}$ -initiated Ras signal transduction pathway. Acta. Pharmacol. Sin., 11, 1155-1160 (2003)
Yue, T. L., Ohlstein, H. E., and Ruffolo, R. R., Apoptosis: a potential target for discovering novel therapies for cardiovascular diseases. Curr. Opin. Chem. Biol., 3, 474-480 (1999)

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