Abstract
Aim:
To investigate the apoptotic mechanism of pseudolaric acid B (PAB) in human breast cancer MCF-7 cells.
Methods:
3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide analysis and morphological changes were applied to detect apoptosis. The percentage of apoptotic and necrotic cells were calculated by the lactate dehydrogenase activity-based cytotoxicity assay, and the protein expression was examined by Western blot analysis.
Results:
PAB and/or the mitogen-activated protein kinases, including p38, c-Jun-N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), did not participate in necrosis. P38 had no obvious function on apoptosis after 4 μmol/L PAB treatment for 36 h, but PAB induced JNK phosphorylation and inhibited ERK phosphorylation in the apoptotic process. In this study the inhibitor of protein tyrosine kinase (PTK) genistein inverted the inhibitory effect of PAB, instead promoting the survival of MCF-7 cells. Like genistein, another PTK inhibitor AG1024 had a similar effect on PAB-treated MCF-7 cells, indicating that PAB activated PTK to induce apoptosis. Together with PAB, genistein increased the expression of p-ERK, and decreased the expressions of JNK and p-JNK in PAB-treated MCF-7 cells at 36 h. And it is considered that the p-ERK and p-JNK were active patterns of ERK and JNK, respectively.
Conclusion:
PTK were upstream of ERK and JNK, and PTK induced apoptosis through activating JNK and inactivating ERK in PAB-treated MCF-7 cells.
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References
Li E, Clark AM, Hufford CD . Antifungal evaluation of PAB, a major constituent of pseudolarix kaempfeAri. 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 cytotoxic principles of Pseudolarix kaempferi: pseudolaric acid-A and B and related derivatives. Planta Med 1990; 56: 383–5.
Yu JH, Cui Q, Jiang YY, Yang W, Tashiro SI, Onodera S, et al. Pseudolaric acid B induces apoptosis, senescence, and mitotic arrest in human breast cancer MCF-7. Acta Pharmacol Sin 2007; 28: 1975–83.
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.
Fadeel B, Orrenius S . Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. J Int Med 2005; 258: 479–517.
Kaufmann SH . Induction of endonucleolytic DNA cleavage in human acute myelogenous breast cancer cells by etoposide, camptothecin, and other cytotoxicanticancer drugs: a cautionary note. Cancer Res 1989; 49: 5870–8.
Rucci N, Recchia I, Angelucci A, Alamanou M, Fattore AD, Fortunati D . Inhibition of protein kinase c-Src reduces the incidence of breast cancer metastases and increases survival in mice: implications for therapy. J Pharmacol Exp Ther 2006; 318: 161–72.
Salvi M, Brunati AM, Toninello A . Tyrosine phosphorylation in mitochondria: A new frontier in mitochondrial signaling. Free Radical Biol Med 2005; 38: 1267–77.
Linassier C, Pierre M, Le PJB, Pierre J .. Mechanisms of action in NIH-3T3 cells of genistein, an inhibitor of EGF receptor tyrosine kinase activity. Biochem Pharmacol 1990; 39: 187–93.
Ullrich A, Schlessinger J . Signal transduction by receptors with tyrosine kinase activity. Cell 1990; 61: 203–12.
Noble MEM, Endicott JA, Johnson LN . Protein kinase inhibitors: insights into drug design from structure. Science 2004; 303: 1800–5.
Kim Hk, Jeong MJ, Kong MY, Han MY, Son KH, Kim HM, et al. Inhibition of Shc/Grb2 protein-protein interaction suppresses growth of B104–1–1 tumors xenografted in nude mice. Life Sci 2005; 78: 321–8.
Conklin CM, Bechberger JF, MacFabe D, Guthrie N, Kurowska EM, Naus CC . Genistein and quercetin increase connexin43 and suppress growth of breast cancer cells. Carcinogenesis 2007; 28: 93–100.
Levitzki A, Gazit A . Tyrosine kinase inhibition: an approach to drug development. Science 1995; 267: 1782–8.
Akiyama T, Ogawara H . Use and specificity of genistein as inhibitor of protein tyrosine kinases. Methods Enzymol 1991; 201: 362–70.
Akiyama T, Ishida J, Nakagawa S, Ogawara H, Watanabe S, Itoh N, et al. Genistein-Induced Changes in Lipid Metabolism of Ovariecto-mized Rats. J Biol Chem 1987; 226: 5592–5.
Schmidt F, Knobbe CB, Frank B, Wolburg H, Weller M . The topoi-somerase II inhibitor, genistein, induces G2/M arrest and apoptosis in human malignant glioma cell lines. Oncol Rep 2008; 19: 1061–6.
Michael H, Alexander H, Andreas PS, Viola B, Detlef S, Hans S . Blockade of IGF-1 receptor tyrosine kinase has antineoplastic effects in hepatocellular carcinoma cells. Biochem Pharmacol 2006; 71: 1435–48.
Klekotka PA, Santoro SA, Wang H, Zutter MM . Specific residues within the alpha 2 integrin subunit cytoplasmic domain regulate migration and cell cycle progression via distinct MAPK pathways. J Biol Chem 2001; 276: 32 353–61.
Schaeffer HJ, Weber MJ . Mitogen-activated protein kinases: specific messages from ubiquitous messengers. Mol Cell Biol 1999; 19: 2435–44.
Xia ZG, Dikens M, Raingesud J, Davis RJ, Greenberg ME . Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 1995; 270: 1336–41.
Hagemann C, Blank JL . The ups and downs of MEK kinase interactions. Cell Signal 2001; 13: 863–75.
Bradham CA, McClay DR . p38 MAPK is essential for secondary axis specification and patterning in sea urchin embryos. Development 2006; 133: 21–32.
Yoon S, Seger R . The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors 2006; 24: 21–44.
Wada T, Penninger JM . Mitogen-activated protein kinases in apoptosis regulation. Oncogene 2004; 23: 2838–49.
Jin Z, El-Deiry WS . Overview of cell death signaling pathways. Cancer Biol Ther 2005; 4: 139–63.
Matsuda S, Gotoh Y, Nishida E . Signaling pathways mediated by the mitogen-activated protein (MAP) kinase/MAP kinase cascade. J Leukoc Biol 1994; 56: 548–53.
Kharbanda S, Ren R, Pandey P, Shafman TD, Feller SM, Weichsel-baum RR, et al. Activation of the c-Abl tyrosine kinase in the stress response to DNA damaging agents. Nature 1995; 376: 785–8.
Kim YM, Talanian RV, Billiar TR . Nitric oxide inhibits apoptosis by preventing increases in caspase-3-like activity via two distinct mechanisms. J Biol Chem 1997; 272: 31 138–48.
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.
Cui Q, Tashiro S, Onodera S, Minami M, Ikejima T . Oridonin induced autophagy in human cervical carcinoma HeLa cells through Ras, JNK, and p38 regulation. J Pharmacol Sci 2007; 105: 317–25.
Yang C, Han R . Genistein suppresses adhesion induced protein tyrosine phosphorylation and invasion of BI6 BL6 melanoma cells. Cancer Lett 1998; 129: 117–25.
Akimoto T, Nonaka T, Ishikawa H, Sakurai H, Saitoh JI, Takahashi T, et al. Genistein, a tyrosine kinase inhibitor, enhanced radiosensitivity in human esophageal cancer cell lines in vitro: possible involvement of inhibition of survival signaltransduction pathways. Int J Radiat Oncol Biol Phys 2001; 50: 195–201.
Tong YG, Zhang XW, Geng MY, Yue JM, Xin XL, Tian F, et al. Pseudolarix acid B, a new tubulin-binding agent, inhibits angiogen-esis 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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Project supported by Shenyang Pharmaceutical University and Showa Pharmaceutical University.
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Yu, Jh., Wang, Hj., Li, Xr. et al. Protein tyrosine kinase, JNK, and ERK involvement in pseudolaric acid B-induced apoptosis of human breast cancer MCF-7 cells. Acta Pharmacol Sin 29, 1069–1076 (2008). https://doi.org/10.1111/j.1745-7254.2008.00835.x
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DOI: https://doi.org/10.1111/j.1745-7254.2008.00835.x
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