Abstract
Objective
To investigate the effect of berbamine on human hepatoma cell line SMMC7721.
Methods
The effects of 24 h and 48 h incubation with different concentrations (0∼64 µg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI (propidium iodide) staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential (Δψ m); the expression of activated caspase3 and caspase9 was analyzed by Western-blot.
Results
The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose-and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential (Δψ m) and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.
Conclusion
Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.
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References
Bhattacharyya, A., Lahiry, L., Mandal, D., Sa, G., Das, T., 2005. Black tea induces tumor cell apoptosis by Bax translocation, loss in mitochondrial transmembrane potential, cytochrome c release and caspase activation. Int. J. Cancer, 117(2):308–315. [doi:10.1002/ijc.21075]
Bossy-Wetzel, E., Green, D.R., 1999. Caspases induce cytochrome c release from mitochondria by activating cytosolic factors. J. Biol. Chem., 274(25):17484–17490. [doi:10.1074/jbc.274.25.17484]
Carnero, A., 2002. Targeting the cell cycle for cancer therapy. Br. J. Cancer, 87(2):129–133. [doi:10.1038/sj.bjc.6600458]
Dong, Q.H., Zheng, S., Xu, R.Z., Lu, Q.H., He, L.M., 2004. Study on effect of berbamine on multidrug resistance leukemia K562/Adr cells. Zhongguo Zhong Xi Yi Jie He Za Zhi, 24(9):820–822 (in Chinese).
Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A., Nagata, S.A., 1998. A caspase-activated DNase that degrades DNA during apoptosis and its inhibitor ICAD. Nature, 391(6662):43–50. [doi:10.1038/34112]
Ge, M.Z., Liu, X., Zhang, Y., 1998. An experiment on immune protection function of berbamine in mice injury by irradiation. Immunol. J., 14:238–240 (in Chinese)
Green, D.R., Reed, J.C., 1998. Mitochondria and apoptosis. Science, 281(5381):1309–1312. [doi:10.1126/science.281.5381.1309]
Hall, A.J., Wild, C.P., 2003. Liver cancer in low and middle income countries. BMJ, 326(7397):994–995. [doi:10.1136/bmj.326.7397.994]
Hirsch, T., Marchetti, P., Susin, S.A., Dallaporta, B., Zamzami, N., Marzo, I., Geuskens, M., Kroemer, G., 1997. The apoptosis-necrosis paradox. Apoptogenic proteases activated after mitochondrial permeability transition determine the model of cell death. Oncogene, 15(13):1573–1581. [doi:10.1038/sj.onc.1201324]
Hsu, Y.L., Kuo, Y.C., Kuo, P.L., Ng, L.T., Kuo, Y.H., Lin, C.C., 2005. Apoptotic effects of extract from Antrodia camphorate fruiting bodies in human hepatocellular carcinoma cell lines. Cancer Lett., 221(1):77–89. [doi:10.1016/j.canlet.2004.08.012]
Koopman, G., Reutelingsperger, C.M.P., Kuijten, G.A.M., Keehnen, R.M.J., Pals, S.T., van Oers, M.T.J., 1994. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood, 84(5):1415–1420.
Küpeli, E., Kosar, M., Yasilada, E., Husnu, K., Baser, C., 2002. A comparative study on the anti-inflammatory, antinociceptive and antipyretic effects of isoquinoline alkaloids from the roots of Turkish Berberis species. Life Sci., 72(6):645–657. [doi:10.1016/S0024-3205(02)02200-2]
Liu, X., Zhou, Z.R., 1996. The effect of berbamine on the immunoregulation of BALB/c mice. J. Chin. Med. Univ., 25:229–231 (in Chinese).
Marchetti, P., Castedo, M., Susin, S.A., Zamzami, N., Hirsch, T., Macho, A., Haeffner, A., Hirsch, F., Geuskens, M., Kroemer, G., 1996. Mitochondrial permeability transition is a central coordinating event of apoptosis. J. Exp. Med., 184(3):1155–1160. [doi:10.1084/jem.184.3.1155]
Pallis, M., Grundy, M., Turzanski, J., Kofler, R., Russell, N., 2001. Mitochondrial membrane sensitivity to depolarization in acute myeloblastic leukemia is associated with spontaneous in vitro apoptosis, wild-type TP53, and vicinal thiol/disulfide status. Blood, 98(2):405–413. [doi:10.1182/blood.V98.2.405]
Parkin, D.M., Bray, F., Ferlay, J., Pisani, P., 2001. Estimating the world cancer burden: GLOBOCAN 2000. Int. J. Cancer, 94(2):153–156. [doi:10.1002/ijc.1440]
PIetenpol, J.A., Stewart, Z.A., 2002. Cell cycle checkpoint signaling: cell cycle arrest versus apoptosis. Toxicology, 181–182:475–481. [doi:10.1016/S0300-483X(02)00460-2]
Schmitt, C.A., Lowe, S.W., 1999. Apoptosis and therapy. J. Pathol., 187(1):127–137. [doi:10.1002/(SICI)1096-9896(199901)187:1〈127::AID-PATH251〉3.0.CO;2-T]
Sun, J.R., Zhang, X.H., He, Z.W., Gu, Y., Yu, Y.Z., Fang, Y.M., Lu, Q.H., Dong, Q.H., Xu, R.Z., 2006. The mechanism of apoptosis of chronic myeloid leukemia cells induced by the novel p210 bcr/abl inhibitor berbamine. Zhonghua Yi Xue Za Zhi, 86(32):2246–2251 (in Chinese).
Thompson, C.B., 1995. Apoptosis in the pathogenesis and treatment of disease. Science, 267(5203):1456–1462. [doi:10.1126/science.7878464]
Thornberry, N.A., Lazebnik, Y., 1998. Caspases: enemies within. Science, 281(5381):1312–1316. [doi:10.1126/science.281.5381.1312]
Wang, X.J., Wei, Y.Q., Yuan, S.L., Liu, G.J., Lu, Y.R., Zhang, J., Wang, W.D., 2005. Potential anticancer activity of tanshinoe IIA against human breast cancer. Int. J. Cancer, 116(5):799–807. [doi:10.1002/ijc.20880]
Wolf, B.B., Schuler, M., Echeverri, F., Green, D.R., 1999. Caspase-3 is the primary activator of apoptotic DNA fragmentation via DNA fragmentation factor-45/inhibitor of caspase-activated DNase inactivation. J. Biol. Chem., 274(43):30651–30656. [doi:10.1074/jbc.274.43.30651]
Wu, D., Lin, M.F., Zhao, X.Y., 2005. Effects of berbamine on K562 cells and its mechanisms in vitro and in vivo. Zhongguo Shi Yan Xue Ye Xue Za Zhi, 13(3):373–378 (in Chinese).
Xu, C.Q., Dong, D.L., Du, Z.M., Chen, D.W., Gong, D.M., Yang, B.F., 2004. Comparison of the anti-arrhythmic effects of matrine and berbamine with amiodarone and RP58866. Yao Xue Xue Bao, 39(9):691–694 (in Chinese).
Xu, R.Z., Dong, Q.H., Yu, Y., Zhao, X., Gan, X., Wu, D., Lu, Q., Xu, X., Yu, X.F., 2006. Berbamine: a novel inhibitor of bcr/abl fusion gene with potent anti-leukemia activity. Leuk. Res., 30(1):17–23. [doi:10.1016/j.leukres.2005.05.023]
Yang, K., Zhao, X., Xiao, P., Liu, G., 1982. Clinical trial of berbamine in 405 leukopenia patients. Yao Hsueh Tung Pao, 17:21–22 (in Chinese).
Zhu, X.W., Sui, W.Z., 1986. Experimental study on the anti-tumor action of berbamine in mice. Zhong Xi Yi Jie He Za Zhi, 6(10):611–613 (in Chinese).
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Project supported by the National Natural Science Foundation of China (No. 30400521), the Science and Technology Department of Zhejiang Province (Nos. 2004D31026 and 2002D3007) and the Education Department of Zhejiang Province (No. 20060427), China
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Wang, Gy., Zhang, Jw., Lü, Qh. et al. Berbamine induces apoptosis in human hepatoma cell line SMMC7721 by loss in mitochondrial transmembrane potential and caspase activation. J. Zhejiang Univ. - Sci. B 8, 248–255 (2007). https://doi.org/10.1631/jzus.2007.B0248
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DOI: https://doi.org/10.1631/jzus.2007.B0248