Pseudolaric Acid B, a Novel Microtubule-Destabilizing Agent That Circumvents Multidrug Resistance Phenotype and Exhibits Antitumor Activity In vivo

Clinical Cancer Research - Tập 11 Số 16 - Trang 6002-6011 - 2005
Vincent Kam Wai Wong1,2, Pauline Chiu1,2, Stephen S.M. Chung3, Larry M. C. Chow4, Yunzhe Zhao4, Burton B. Yang5, Ben C.B. Ko1,2
11Department of Chemistry,
22Open Laboratory of Chemical Biology of the Institute of Molecular Technology for Drug Discovery and Synthesis, and
33Institute of Molecular Biology, The University of Hong Kong;
44Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hong Kong Special Administrative Region, China; and
55Sunnybrook and Women's College Health Sciences Centre and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada

Tóm tắt

Abstract Purpose: Pseudolaric acid B (PAB) is the major bioactive constituent in the root bark of Pseudolarix kaempferi that has been used as an antifungal remedy in traditional Chinese medicine. Previous studies showed that PAB exhibited substantial cytotoxicity. The aims of this study were to elucidate the molecular target of PAB, to examine its mechanism of action, and to evaluate the efficacy of this compound in vivo. Experimental Design: The effect of PAB on cell growth inhibition toward a panel of cancer cell lines was assayed. Cell cycle analysis, Western blotting, immunocytochemistry, and apoptosis analysis were carried out to examine the mechanism of action. Tubulin polymerization assays were conducted to examine the interaction between PAB and tubulin. A P-glycoprotein–overexpressing cell line was used to evaluate the efficacy of PAB toward multidrug-resistant phenotypes. In vivo efficacy of PAB was evaluated by the murine xenograft model. Results: PAB induces cell cycle arrest at G2-M transition, leading to apoptosis. The drug disrupts cellular microtubule networks and inhibits the formation of mitotic spindles. Polymerization of purified bovine brain tubulin was dose-dependently inhibited by PAB. Furthermore, PAB circumvents the multidrug resistance mechanism, displaying notable potency also in P-glycoprotein–overexpressing cells. Finally, we showed that PAB is effective in inhibiting tumor growth in vivo. Conclusions: We identified the microtubules as the molecular target of PAB. Furthermore, we showed that PAB circumvents P-glycoprotein overexpression-induced drug resistance and is effective in inhibiting tumor growth in vivo. Our work will facilitate the future development of PAB as a cancer therapeutic.

Từ khóa


Tài liệu tham khảo

Newman DJ, Cragg GM, Snader KM. Natural products as sources of new drugs over the period 1981-2002. J Nat Prod 2003;66:1022–37.

Breinbauer R, Vetter IR, Waldmann H. From protein domains to drug candidates—natural products as guiding principles in the design and synthesis of compound libraries. Angew Chem Int Ed Engl 2002;41:2878.

Mann J. Natural products in cancer chemotherapy: past, present and future. Nat Rev Cancer 2002;2:143–8.

Giannakakou P, Sackett D, Fojo T. Tubulin/microtubules: still a promising target for new chemotherapeutic agents. J Natl Cancer Inst 2000;92:182–3.

Downing KH, Nogales E. Tubulin structure: insights into microtubule properties and functions. Curr Opin Struct Biol 1998;8:785–91.

Hamel E. Antimitotic natural products and their interactions with tubulin. Med Res Rev 1996;16:207–31.

Jordan MA, Wilson L. Microtubules as a target for anticancer drugs. Nat Rev Cancer 2004;4:253–65.

Tozer GM, Kanthou C, Parkins CS, Hill SA. The biology of the combretastatins as tumour vascular targeting agents. Int J Exp Pathol 2002;83:21–38.

Giannakakou P, Villalba L, Li H, Poruchynsky M, Fojo T. Combinations of paclitaxel and vinblastine and their effects on tubulin polymerization and cellular cytotoxicity: characterization of a synergistic schedule. Int J Cancer 1998;75:57–63.

Li E, Clark AM, Hufford CD. Antifungal evaluation of pseudolaric acid B, a major constituent of Pseudolarix kaempferi. J Nat Prod 1995;58:57–67.

Zhou BN. Some progress on the chemistry of natural bioactive terpenoids from Chinese medicinal plants. Mem Inst Oswaldo Cruz 1991;86 Suppl 2:219–26.

Wang WC, Lu RF, Zhao SX, Zhu YZ. Antifertility effect of pseudolaric acid B. Zhongguo Yao Li Xue Bao 1982;3:188–92.

Pan DJ, Li ZL, Hu CQ, Chen K, Chang JJ, Lee KH. The cytotoxic principles of Pseudolarix kaempferi: pseudolaric acid-A and -B and related derivatives. Planta Med 1990;56:383–5.

Zhou BN, Ying BP, Song GQ, Chen ZX, Han J, Yan YF. Pseudolaric acids from Pseudolarix kaempferi. Planta Med 1983;47:35–8.

Zhou J, Cheng SC, Luo D, Xie Y. Study of multi-drug resistant mechanisms in a taxol-resistant hepatocellular carcinoma QGY-TR 50 cell line. Biochem Biophys Res Commun 2001;280:1237–42.

Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res 1987;47:943–6.

Teramoto H, Malek RL, Behbahani B, Castellone MD, Lee NH, Gutkind JS. Identification of H-Ras, RhoA, Rac1 and Cdc42 responsive genes. Oncogene 2003;22:2689–97.

Koizumi Y, Arai M, Tomoda H, Omura S. Oxaline, a fungal alkaloid, arrests the cell cycle in M phase by inhibition of tubulin polymerization. Biochim Biophys Acta 2004;1693:47–55.

Guba M, von Breitenbuch P, Steinbauer M, et al. Rapamycin inhibits primary and metastatic tumor growth by antiangiogenesis: involvement of vascular endothelial growth factor. Nat Med 2002;8:128–35.

Wu Y, Sheng W, Chen L, et al. Versican V1 isoform induces neuronal differentiation and promotes neurite outgrowth. Mol Biol Cell 2004;15:2093–104.

Sheng W, Wang G, Wang Y, et al. The roles of versican V1 and V2 isoforms in cell proliferation and apoptosis. Mol Biol Cell 2005.

Loike JD, Horwitz SB. Effects of podophyllotoxin and VP-16-213 on microtubule assembly in vitro and nucleoside transport in HeLa cells. Biochemistry 1976;15:5435–43.

Clifford B, Beljin M, Stark GR, Taylor WR. G2 arrest in response to topoisomerase II inhibitors: the role of p53. Cancer Res 2003;63:4074–81.

Smits VA, Klompmaker R, Vallenius T, Rijksen G, Makela TP, Medema RH. p21 inhibits Thr161 phosphorylation of Cdc2 to enforce the G2 DNA damage checkpoint. J Biol Chem 2000;275:30638–43.

Jordan MA. Mechanism of action of antitumor drugs that interact with microtubules and tubulin. Curr Med Chem Anti-Canc Agents 2002;2:1–17.

Downing KH. Structural basis for the interaction of tubulin with proteins and drugs that affect microtubule dynamics. Annu Rev Cell Dev Biol 2000;16:89–111.

Ravelli RB, Gigant B, Curmi PA, et al. Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain. Nature 2004;428:198–202.

Ling V, Charles F. Kettering Prize. P-glycoprotein and resistance to anticancer drugs. Cancer 1992;69:2603–9.

Ueda K, Cardarelli C, Gottesman MM, Pastan I. Expression of a full-length cDNA for the human “MDR1” gene confers resistance to colchicine, doxorubicin, and vinblastine. Proc Natl Acad Sci U S A 1987;84:3004–8.

Leonessa F, Green D, Licht T, et al. MDA435/LCC6 and MDA435/LCC6MDR1: ascites models of human breast cancer. Br J Cancer 1996;73:154–61.

Chaudhuri AR, Luduena RF. Griseofulvin: a novel interaction with bovine brain tubulin. Biochem Pharmacol 1996;51:903–9.

Pettit RK, McAllister SC, Pettit GR, Herald CL, Johnson JM, Cichacz ZA. A broad-spectrum antifungal from the marine sponge Hyrtios erecta. Int J Antimicrob Agents 1997;9:147–52.

Uckun FM, Mao C, Jan ST, et al. Spongistatins as tubulin targeting agents. Curr Pharm Des 2001;7:1291–6.

Stephen RL, Gustafsson MC, Jarvis M, et al. Activation of peroxisome proliferator-activated receptor δ stimulates the proliferation of human breast and prostate cancer cell lines. Cancer Res 2004;64:3162–70.

Mitchison T, Kirschner M. Dynamic instability of microtubule growth. Nature 1984;312:237–42.

Rusan NM, Fagerstrom CJ, Yvon AM, Wadsworth P. Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-α tubulin. Mol Biol Cell 2001;12:971–80.

Mitchison TJ. Microtubule dynamics and kinetochore function in mitosis. Annu Rev Cell Biol 1988;4:527–49.

Dumontet C, Sikic BI. Mechanisms of action of and resistance to antitubulin agents: microtubule dynamics, drug transport, and cell death. J Clin Oncol 1999;17:1061–70.

Geney R, Ungureanu M, Li D, Ojima I. Overcoming multidrug resistance in taxane chemotherapy. Clin Chem Lab Med 2002;40:918–25.

Ambudkar SV, Kimchi-Sarfaty C, Sauna ZE, Gottesman MM. P-glycoprotein: from genomics to mechanism. Oncogene 2003;22:7468–85.

Mabjeesh NJ, Escuin D, La Vallee TM, et al. 2ME2 inhibits tumor growth and angiogenesis by disrupting microtubules and dysregulating HIF. Cancer Cell 2003;3:363–75.

Pribluda VS, Gubish ER Jr., Lavallee TM, Treston A, Swartz GM, Green SJ. 2-Methoxyestradiol: an endogenous antiangiogenic and antiproliferative drug candidate. Cancer Metastasis Rev 2000;19:173–9.

Petitclerc E, Deschesnes RG, Cote MF, et al. Antiangiogenic and antitumoral activity of phenyl-3-(2-chloroethyl)ureas: a class of soft alkylating agents disrupting microtubules that are unaffected by cell adhesion-mediated drug resistance. Cancer Res 2004;64:4654–63.

Micheletti G, Poli M, Borsotti P, et al. Vascular-targeting activity of ZD6126, a novel tubulin-binding agent. Cancer Res 2003;63:1534–7.

Li MH, Miao ZH, Tan WF, et al. Pseudolaric acid B inhibits angiogenesis and reduces hypoxia-inducible factor 1α by promoting proteasome-mediated degradation. Clin Cancer Res 2004;10:8266–74.

Thông tin
Thông tin xuất bản

Clinical Cancer Research

Tập 11 Số 16

6002-6011

Thông tin tác giả