Full Text:   <1287>

CLC number: R55

On-line Access: 2011-07-04

Received: 2010-06-18

Revision Accepted: 2010-12-27

Crosschecked: 2011-06-12

Cited: 8

Clicked: 4276

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.7 P.568-574

10.1631/jzus.B1000230


Berbamine inhibits proliferation and induces apoptosis of KU812 cells by increasing Smad3 activity


Author(s):  Yun Liang, Xi Qiu, Rong-zhen Xu, Xiao-ying Zhao

Affiliation(s):  Department of Hematology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China, Cancer Institute, School of Medicine, Zhejiang University, Hangzhou 310009, China

Corresponding email(s):   zrxz@zju.edu.cn

Key Words:  Berbamine, Apoptosis, Smad3, KU812 cells


Yun Liang, Xi Qiu, Rong-zhen Xu, Xiao-ying Zhao. Berbamine inhibits proliferation and induces apoptosis of KU812 cells by increasing Smad3 activity[J]. Journal of Zhejiang University Science B, 2011, 12(7): 568-574.

@article{title="Berbamine inhibits proliferation and induces apoptosis of KU812 cells by increasing Smad3 activity",
author="Yun Liang, Xi Qiu, Rong-zhen Xu, Xiao-ying Zhao",
journal="Journal of Zhejiang University Science B",
volume="12",
number="7",
pages="568-574",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000230"
}

%0 Journal Article
%T Berbamine inhibits proliferation and induces apoptosis of KU812 cells by increasing Smad3 activity
%A Yun Liang
%A Xi Qiu
%A Rong-zhen Xu
%A Xiao-ying Zhao
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 7
%P 568-574
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000230

TY - JOUR
T1 - Berbamine inhibits proliferation and induces apoptosis of KU812 cells by increasing Smad3 activity
A1 - Yun Liang
A1 - Xi Qiu
A1 - Rong-zhen Xu
A1 - Xiao-ying Zhao
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 7
SP - 568
EP - 574
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000230


Abstract: 
Objective: The cytotoxic effect of berbamine on chronic myeloid leukemia (CML) cell line KU812 was evaluated, and the mechanisms of its action were explored. Methods: The effect of berbamine on the KU812 cell growth was determined by methyl thiazolyl tetrazolium (MTT) assay. Flow cytometry was used to profile cell cycle alteration upon berbamine treatment. Reverse transcription polymerase chain reaction (RT-PCR) was carried out to determine the transcripts of transforming growth factor-β (TGF-β) receptors (TβRs), smad3, c-Myc, cyclin D1, p21Cip1(p21), and p27Kip1(p27). Changes in the protein levels of total smad3, phosphorylated smad3, the downstream targets of smad3, and specific apoptosis-related factors were evaluated by Western blotting. Results: berbamine inhibited KU812 cell proliferation in a dose- and time-dependent manner, and the half maximal inhibitory concentration (IC50) values for treatments of 24, 48, and 72 h were 5.83, 3.43, and 0.75 μg/ml, respectively. berbamine induced G1 arrest as well as apoptosis in KU812 cells. Transcriptions of smad3 and p21 were up-regulated, while those of TβRI, TβRII, c-Myc, cyclin D1 and p27 were not changed significantly. The protein levels of both total smad3 and phosphorylated smad3 were both up-regulated after berbamine treatment, together with decreased c-Myc and cyclin D1 and increased p21. Meanwhile, the levels of the anti-apoptotic proteins, such as Bcl-2 and Bcl-xL, were decreased, whereas pro-apoptotic Bax was increased. Conclusions: berbamine suppresses KU812 cell proliferation through induction of cell cycle arrest in G1 and apoptosis. It activates smad3 without additional stimulation of TGF-β, and alters the levels of the smad3 downstream targets, including c-Myc, cyclin D1 and p21. Our findings suggest that berbamine is a promising drug in the treatment of advanced stage patients with CML.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]An, X., Tiwari, A.K., Sun, Y., Ding, P.R., 2010. BCR-ABL tyrosine kinase inhibitors in the treatment of Philadelphia chromosome positive chronic myeloid leukemia: a review. Leuk. Res., 34(10):1255-1268.

[2]Chaudhury, A., Howe, P.H., 2009. The tale of transforming growth factor-β (TGFβ) signaling: a soigné enigma. IUBMB Life, 61(10):929-939.

[3]Claassen, G.F., Hann, S.R., 2000. A role for transcriptional repression of p21CIP1 by c-Myc in overcoming transforming growth factor β-induced cell-cycle arrest. PNAS, 97(17):9498-9503.

[4]Dong, M., Blobe, G.C., 2006. Role of transforming growth factor-β in hematologic malignancies. Blood, 107(12):4589-4596.

[5]Druker, B.J., Guilhot, F., O′Brien, S.G., Gathmann, I., Kantarjian, H., Gattermann, N., Deininger, M.W., Silver, R.T., Goldman, J.M., Stone, R.M., et al., 2006. Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N. Engl. J. Med., 355(23):2408-2417.

[6]Elliott, R.L., Blobe, G.C., 2005. Role of transforming growth factor β in human cancer. J. Clin. Oncol., 23(9):2078-2093.

[7]Engelman, J.A., Settleman, J., 2008. Acquired resistance to tyrosine kinase inhibitors during cancer therapy. Curr. Opin. Genet. Dev., 18(1):73-79.

[8]Kim, S.J., Letterio, J., 2003. Transforming growth factor-β signaling in normal and malignant hematopoiesis. Leukemia, 17(9):1731-1737.

[9]Liang, Y., Xu, R.Z., Zhang, L., Zhao, X.Y., 2009. Berbamine, a novel nuclear factor κB inhibitor, inhibits growth and induces apoptosis in human myeloma cells. Acta Pharmacol. Sin., 30(12):1659-1665.

[10]Mitani, K., 2004. Molecular mechanisms of leukemogenesis by AML1/EVI-1. Oncogene, 23(24):4263-4269.

[11]Ozaki, M., Kanemitsu, N., Yasukawa, M., Fujita, S., 1989. Basophilic crisis of chronic myelogenous leukemia. Jpn. J. Med., 28(1):67-71.

[12]Pardali, K., Kowanetz, M., Heldin, C.H., Moustakas, A., 2005. Smad pathway-specific transcriptional regulation of the cell cycle inhibitor p21(WAF1/Cip1). J. Cell. Physiol., 204(1):260-272.

[13]Richter, B.W.M., Duckett, C.S., 2000. The IAP proteins: caspase inhibitors and beyond. Sci. STKE, 44:PE1.

[14]Rooke, H.M., Vitas, M.R., Crosier, P.S., Crosier, K.E., 1999. The TGF-β type II receptor in chronic myeloid leukemia: analysis of microsatellite regions and gene expression. Leukemia, 13(4):535-541.

[15]Ruscetti, F.W., Akel, S., Bartelmez, S.H., 2005. Autocrine transforming growth factor-β regulation of hematopoiesis: many outcomes that depend on the context. Oncogene, 24(37):5751-5763.

[16]Steinberg, M., 2007. Dasatinib: a tyrosine kinase inhibitor for the treatment of chronic myelogenous leukemia and philadelphia chromosome-positive acute lymphoblastic leukemia. Clin. Ther., 29(11):2289-2308.

[17]Wei, Y.L., Xu, L., Liang, Y., Xu, X.H., Zhao, X.Y., 2009a. Berbanine exhibits potent antitumor effects on imatinib-resistant CML cells in vitro and in vivo. Acta Pharmacol. Sin., 30(4):451-457.

[18]Wei, Y.L., Xu, L., Zhao, X.Y., 2009b. Mechanism related to inhibition of leukemia K562 cells by berbamine. J. Zhejiang Univ. (Med. Sci.), 38(4):387-391 (in Chinese).

[19]Wolfraim, L.A., Fernandez, T.M., Mamura, M., Fuller, W.L., Kumar, R., Cole, D.E., 2004. Loss of Smad3 in acute T-cell lymphoblastic leukemia. N. Engl. J. Med., 351(6):552-559.

[20]Xu, R.Z., Dong, Q.H., Yu, Y.Z., Zhao, X.Y., Gan, X.X., Wu, D., Lu, Q.H., Xu, X.H., Yu, X.F., 2006. Berbanine: a novel inhibitor of bcr/abl fusion gene with potent anti-leukemia activity. Leuk. Res., 30(1):17-23.

[21]Yagi, K., Furuhashi, M., Aoki, H., Goto, D., Kuwano, H., Sugamura, K., Miyazono, K., Mitsuyasu Kato, M., 2002. C-Myc is a downstream target of the Smad pathway. J. Biol. Chem., 277(1):854-861.

[22]Zhang, L., Sun, Z., Shen, A.L., Ma, L., Jiang, X.Y., Ma, G.J., 2003. The effects of the Smad3-knockout on the hematopoiesis of mouse. Chin. J. Biotechnol., 19(4):428-432 (in Chinese).

[23]Zhu, H.J., Burgess, A.W., 2001. Regulation of transforming growth factor-β signaling. Mol. Cell Biol. Res. Commun., 4(6):321-330.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

王巨存@天津市天津医院<wangjucun2000@163.com>

2011-11-10 12:58:00

I am very intrested in your study field. I think you did great and meaningful job.

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE