Abstract
Aim:
The aim of the present study was to investigate the inhibitory effect of pseudolaric acid B (PAB) on human breast cancer MCF-7 cells.
Methods:
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis, morphological changes, acridine orange staining, and agarose gel electrophoresis were applied to detect apoptosis. The percentage of apoptotic and necrotic cells was calculated by the lactate dehydrogenase activity-based cytotoxicity assay; senescence associated (SA)-β-galactosidase activity was detected to evaluate senescence; flow cytometric analysis of propidium iodide staining was carried out to investigate the distribution of cell cycle, and the protein expression was examined by Western blot analysis.
Results:
During apoptosis, the half maximal inhibitory concentration IC50 was 3.4 and 1.35 μmol/L at 36 and 48 h after PAB treatment, respectively. The MCF-7 cells exposed to PAB showed typical characteristics of apoptosis, including the morphological changes and DNA fragmentation. The MCF-7 cells treated with 4 μmol/L PAB for 36 h underwent apoptosis, but not necrosis. The apoptosis induced by PAB was independent of the death receptor pathway. The senescent cells became larger and flatter, and the SA-β-galactosidase staining was positive. PAB induced obvious mitotic arrest and it preceded apoptosis and senescence. The expressions of p21 and p53 was upregulated with PAB treatment, and cyclin B1 was upregulated and transported from the cytoplasm to nuclei, and sustained stable levels.
Conclusion:
PAB induced mitotic arrest in the MCF-7 cells and inhibited proliferation through apoptosis and senescence. The apoptosis was independent of the death receptor pathway.
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References
Li E, Clark AM, Hufford CD . Antifungal evaluation of PAB, a major constituent of Pseudolarix kaempferi. J Nat Prod 1995; 58: 57–67.
Wang WC, Lu RF, Zhao SX, Gu ZP . Comparison of early pregnancy-terminating effect and toxicity between pseudolaric acids A and B. Acta Pharmacol Sin 1988; 9: 445–8.
Wang WC, Lu RF, Zhao SX, Zhu YZ . Antifertility effect of PAB. Acta Pharmacol Sin 1982; 3: 188–92.
Pan DJ, Li ZL, Hu CQ, Chen K, Chang JJ, Lee KH . The cyto-toxic principles of Pseudolarix Kaempferi: Pseudolaric acid-A and B and related derivatives. Planta Med 1990; 56: 383–5.
Gong XF, Wang MW, Tashiro SI, Onodera S, Ikejima T . PAB induces apoptosis through P53 and Bax/Bcl-2 pathways in human melanoma A375-S2 cell. Arch Pham Res 2005; 28: 68–72.
Zhang M, Mo X, Chen XY, Chen L, Li LZ . Studies of apoptosis on K562 cell line induced by pseudolaricd acid B in vitro. Chin Tradit Herb Drugs 2002; 33: 533–5.
Gong XF, Wang MW, Tashiro SI, Onodera S, Ikejima T . Involvement of JNK-initiated p53 accumulation and phosphorylation of p53 in pseudolaric acid B induced cell death. Exp Mol Med 2006; 38: 428–34.
Campisi J . Cellular senescence as a tumor-suppressor mechanism. Trends Cell Biol 2001; 11: S27–31.
Rebbaa A, Zheng X, Chou PM, Mirkin BL . Caspase inhibition switches doxorubicin-induced apoptosis to senescence. Oncogene 2003; 22: 2805–11.
Hartwell LH, Weinert TA . Checkpoints: controls that ensure the order of cell cycle events. Science 1989; 246: 629–34.
Pietenpol JA, Stewart ZA . Cell cycle checkpoint signaling: cell cycle arrest versus apoptosis. Toxicology 2002; 181–182: 475–81.
Micco RD, Fumagalli M . Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 2006, 444: 638–42.
Sherr CJ, Roberts JM . Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev 1995; 9: 1149–63.
Caroline CR, Philippe R, Elisheva YR . Mechanisms of P53-induced apoptosis: in search of genes which are regulated during P53-mediated cell death. Toxicol Lett 1998; 102–103: 491–6.
Peng CY, Graves PR, Thoma RS, Wu ZQ, Shaw AS, Piwnica-Worms H . Mitotic and G2 checkpoint control: regulation of 14-3-3 protein binding by phosphorylation of Cdc25C on serine-216. Science 1997; 277: 1501–5.
Murray AW, Kirschner MW . Dominoes and clocks: the union of two views of cell cycle regulation. Science 1989; 246: 614–21.
Sarin A, Hadid E, Henkart PA . Caspase dependence of target cell damage induced by cytotoxic T lymphocytes. J Immunol 1998; 161: 2810–3.
Kim YM, Talanian RV, Billiar TR . Nitric oxide inhibits apoptosis by preventing increases on caspase-3-like activity via two distinct mechanisms. J Biol Chem 1997; 272: 31 138–47.
Dimri G P, Leet X, Basile G, Acost M, Scorrt G, Campisi J, et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Sci Acad USA 1995; 92: 9363–7.
Lee KY, Park JA, Chung E, Lee YH, Kim SI, Lee SK . Ginsenoside Rh2 blocks the cell cycle of SK-KEP-1cells at the G1/S boundary by selectively inducing the protein expression of p27kipl. Cancer Lett 1996; 110: 193–200.
Nalini K, Rayudu G, Vsha G, Jinah C, Henry JF . Role of protein kinase C in basal and hydrogen peroxide stimulated NF-κB activation in the murine macrophage. Arch Biochem Biochem Biophys 1998; 350: 79–86.
Janicke RU, Sprengart ML, Wati MR, Porter AG . Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 1998; 273: 9357–60.
Jin P, Hardy S, Morgan DO . Nuclear localization of cyclin B1 controls mitotic entry after DNA damage. J Cell Biol 1998; 141: 875–85.
Kerr JF, Wyllie AH, Currie AR . Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972; 26: 239–57.
Cui Q, Yu JH, Wu JN, Tashiro S, Onodera S, Minami M, et al. P53-mediated cell cycle arrest and apoptosis through a caspase-3-independent, but caspase-9-dependent pathway in oridonin-treated MCF-7 human breast cancer cells. Acta Pharmacol Sin 2007; 28: 1057–66.
Cregan SP, Dawson VL, Slack RS . Role of AIF in caspase-depen-dent and caspase-independent cell death. Oncogene 2004; 23: 2785–96.
Chinnaiyan AM, O'Rourke K, Tewari M, Dixit VM . FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell 1995; 81: 505–12.
May P, May E . Twenty years of p53 research: structural and functional aspects of the p53 protein. Oncogene 1999; 18: 7621–36.
Macleod KF, Sherry N, Hannon G, Beach D, Tokino T, Kinzler K, et al. P53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage. Genes Dev 1995; 9: 935–44.
Flatt PM, Tang LJ, Scatena CD, Szak ST, Pietenpol JA . P53 regulation of G2 checkpoint is retinoblastoma protein dependent. Mol Cell Biol 2000; 20: 4210–23.
Ingber DE, Prusty D, Sun Z, Betensky H, Wang N . Cell shape, cytoskeletal mechanics and cell cycle control in angiogenesis. J Biomech 1995; 28: 1471–84.
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.
Wang LG, Liu XM, Kreis W, Budman DR . The effect of antimicrotubule agents on signal transduction pathways of apoptosis: a review. Cancer Chemother Pharmacol 1999; 44: 355–61.
Tong YG, Zhang XW, Geng MY, Yue JM, Xin XL, Tian F, et al. Pseudolarix acid B, a new tubulin-binding agent, inhibits angio-genesis by interacting with a novel binding site on tubulin. Mol Pharmacol 2006; 69: 1226–33.
Wong VK, Chiu P, Chung SS, Chow LM, Zhao YZ, Yang BB, et al. Pseudolaric acid B, a novel microtubule-destabilizing agent that circumvents multidrug resistance phenotype and exhibits antitumor activity in vivo. Clin Cancer Res 2005; 11: 6002–11.
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Yu, Jh., Cui, Q., Jiang, Yy. et al. Pseudolaric acid B induces apoptosis, senescence, and mitotic arrest in human breast cancer MCF-7. Acta Pharmacol Sin 28, 1975–1983 (2007). https://doi.org/10.1111/j.1745-7254.2007.00706.x
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DOI: https://doi.org/10.1111/j.1745-7254.2007.00706.x
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