Abstract
Aim:
To verify the suppressive effect of berberine on the proliferation of the human pulmonary giant cell carcinoma cell line PG and to demonstrate the mechanisms behind the antitumoral effects of berberine.
Methods:
The proliferative effects of PG cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetry. The cell cycle was examined by flow cytometry. The expression level of cyclin D1 was detected by RT-PCR. The activities of the activating protein-1 (AP-1) and NF-κB signaling pathways related to cyclin D1 were examined by luciferase assay. The cytoplasmic level of c-Jun was detected by Western blot analysis. An electrophoretic mobility shift assay was used to examine the binding of transcription factors to the cyclin D1 gene (CCND1) AP-1 motif.
Results:
The results showed that the proliferation of PG cells treated with different concentrations (10, 20, and 40 μg/mL) of berberine for 24 and 48 h was suppressed significantly compared to the control group. After treatment with berberine, the proportion of PG cells at the G0/G1 phase increased, while cells at the S and G2/M phases decreased. Berberine could inhibit the expression of cyclin D1 in PG cells. Berberine inhibited the activity of the AP-1 signaling pathway, but had no significant effect on the NF-κB signaling pathway. Berberine suppressed the expression of c-Jun and decreased the binding of transcription factors to the CCND1 AP-1 motif.
Conclusion:
Berberine suppresses the activity of the AP-1 signaling pathway and decreases the binding of transcription factors to the CCND1 AP-1 motif. This is one of the important mechanisms behind the antitumoral effects of berberine as a regulator of cyclin D1.
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Xu LH, Liu Y, He XH . Inhibitory effects of berberine on the activation and cell cycle progression of human peripheral lymphocytes. Cell Mol Immunol 2005; 2: 295–300.
Zhao HP, Hong Y, Xie JD, Xie XR, Wang J, Fan JB . Effect of berberine on left ventricular remodeling in renovascular hypertensive rats. Yao Xue Xue Bao 2007; 42: 336–41.
Letasiova S, Jantova S, Miko M, Ovadekova R, Horvathova M . Effect of berberine on proliferation, biosynthesis of macromolecules, cell cycle and induction of intercalation with DNA, dsDNA damage and apoptosis in Ehrlich ascites carcinoma cells. J Pharm Pharmacol 2006; 58: 263–70.
Peng PL, Hsieh YS, Wang CJ, Hsu JL, Chou FP . Inhibitory effect of berberine on the invasion of human lung cancer cells via decreased productions of urokinase-plasminogen activator and matrix metalloproteinase-2. Toxicol Appl Pharmacol 2006; 214: 8–15.
Hao Y, Xu BW, Zheng H, Wang Q, Qiu QY, Huang QF . Effect of berberine on invasion and migration of PG cells from a high metastatic human giant lung carcinoma cell line. Chin J Pathophysiol 2007; 23: 474–8.
Matsushime H, Roussel MF, Ashmun RA, Sherr CJ . Colony-stimulating factor 1 regulates novel cyclins during the G1 phase of the cell cycle. Cell 1991; 65: 701–13.
Kato JY, Matsuoka M, Strom DK, Sherr CJ . Regulation of cyclin D dependent kinase 4 (cdk4) by cdk4-activating kinase. Mol Cell Biol 1994; 14: 2713–21.
Kato J, Matsushime H, Hiebert SW, Ewen ME, Sherr CJ . Direct binding of cyclin D to the retinoblastoma gene product (pRb) and pRb phosphorylation by the cyclin D-dependent kinase CDK4. Genes Dev 1993; 7: 331–42.
Gautschi O, Ratschiller D, Gugger M, Betticher DC, Heighway J . Cyclin D1 in non-small cell lung cancer: a key driver of malignant transformation. Lung Cancer 2007; 55: 1–14.
Eferl R, Wagner EF . AP-1: a double-edged sword in tumorigenesis. Nat Rev Cancer 2003; 3: 859–68.
Joyce D, Bouzahzah B, Fu M, Albanese C, D'Amico M, Steer J, et al. Integration of rac-dependent regulation of cyclin D1 transcription through a nuclear factor-κB-dependent pathway. J Biol Chem 1999; 274: 25245–9.
Natsume H, Sasaki S, Kitagawa M, Kashiwabara Y, Matsushita A, Nakano K, et al. β-Catenin/Tcf-1-mediated transactivation of cyclin D1 promoter is negatively regulated by thyroid hormone. Biochem Biophys Res Commun 2003; 309: 408–13.
Shaulian E, Karin M . AP-1 in cell proliferation and survival. Oncogene 2001; 20: 2390–400.
Fu M, Wang C, Zhang X, Pestell RG . Signal transduction inhibitors in cellular function. Methods Mol Biol 2004; 284: 15–36.
Wu BQ, Sun YK, Zheng J, Fang WG, Wang JL . Metastatic behavior of cultured human lung carcinoma cell lines in nude mice. In: Rygaard J, Brünner N, Græm N, Spang-Thomsen M . Immunedeficient animals in biomedical research. Proceedings of the 5th International Workshop on Immune-deficient Animals; Copenhagen, Denmark, Oct 13–16, 1985; Karger, Basel, Switzerland; 20 May 1987; p 296–302.
Xia D, Li X, Lou Y, Han W, Ding P, Zhang Y, et al. Overexpression of chemokine-like factor 2 promotes the proliferation and survival of CaCl2 skeletal muscle cells. Biochim Biophys Acta 2002; 1591: 163–73.
Albanese C, Johnson J, Watanabe G, Eklund N, Vu D, Arnold A, et al. Transforming p21ras mutants and c-Ets-2 activate the cyclin D1 promoter through distinguishable regions. J Biol Chem 1995; 270: 23589–97.
Yang-Yen HF, Chambard JC, Sun YL, Smeal T, Schmidt TJ, Drouin J, et al. Transcriptional interference between c-Jun and the glucocorticoid receptor: mutual inhibition of DNA binding due to direct protein-protein interaction. Cell 1990; 62: 1205–15.
Lin S, Tsai SC, Lee CC, Wang BW, Liou JY, Shyu KG . Berberine inhibits HIF-1α expression via enhanced proteolysis. Mol Pharmacol 2004; 66: 612–9.
Jantova S, Cipak L, Letasiova S . Berberine induces apoptosis through a mitochondrial/caspase pathway in human promonocytic U937 cells. Toxicol In Vitro 2007; 21: 25–31.
Lin CC, Lin SY, Chung JG, Lin JP, Chen GW, Kao ST . Down-regulation of cyclin B1 and up-regulation of wee1 by berberine promotes entry of leukemia cells into the G2/M-phase of the cell cycle. Anticancer Res 2006; 26: 1097–104.
Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Akao S, Fujiwara H . Inhibition of activator protein 1 activity by berberine in human hepatoma cells. Planta Med 1999; 65: 381–3.
Mitani N, Murakami K, Yamaura T, Ikeda T, Saiki I . Inhibitory effect of berberine on the mediastinal lymph node metastasis produced by orthotopic implantation of Lewis lung carcinoma. Cancer Lett 2001; 165: 35–42.
Lee RJ, Albanese C, Fu M, D'Amico M, Lin B, Watanabe G, et al. Cyclin D1 is required for transformation by activated Neu and is induced through an E2F-dependent signaling pathway. Mol Cell Biol 2000; 20: 672–83.
Fantl V, Stamp G, Andrews A, Rosewell I, Dickson C . Mice lacking cyclin D1 are small and show defects in eye and mammary gland development. Genes Dev 1995; 9: 2364–72.
Yu Q, Geng Y, Sicinski P . Specific protection against breast cancers by cyclin D1 ablation. Nature 2001; 411: 1017–21.
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Project supported by the National High Technology Research and Development Program of China (863 Program; No 2006AA02A305).
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Luo, Y., Hao, Y., Shi, Tp. et al. Berberine inhibits cyclin D1 expression via suppressed binding of AP-1 transcription factors to CCND1 AP-1 motif. Acta Pharmacol Sin 29, 628–633 (2008). https://doi.org/10.1111/j.1745-7254.2008.00786.x
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DOI: https://doi.org/10.1111/j.1745-7254.2008.00786.x
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