The objectives of this study were to evaluate the effects of tanshinones from a Chinese herb Salvia Miltiorrhiza on the growth of breast cancer cells, and to elucidate cellular and molecular mechanisms of action. Tanshinones showed the dose-dependent effect on the growth inhibition of breast cancer cells in vitro, with tanshinone I (T1) the most potent agent. T1 was also the only tanshinone to have potent activity in inhibiting the growth of the triple-negative breast cancer cell line MDA-MB231. T1 caused cell cycle arrests of both estrogen-dependent and estrogen-independent cell lines associated with alterations of cyclinD, CDK4 and cyclinB, and induced breast cancer cell apoptosis associated with upregulation of c-PARP and downregulation of survivin and Aurora A. Among these associated biomarkers, Aurora A showed the most consistent pattern with the anti-growth activity of tanshinones. Overexpression of Aurora A was also verified in breast tumors. The gene function assay showed that knockdown of Aurora A by siRNA dramatically reduced the growth-inhibition and apoptosis-induction activities of T1, suggesting Aurora A as an important functional target of T1 action. On the other hand, tanshinones had much less adverse effects on normal mammary epithelial cells. Epigenetic mechanism studies showed that overexpression of Aurora A gene in breast cancer cells was not regulated by gene promoter DNA methylation, but by histone acetylation. T1 treatment significantly reduced acetylation levels of histone H3 associated with Aurora A gene. Our results supported the potent activity of T1 in inhibiting the growth of breast cancer cells in vitro in part by downregulation of Aurora A gene function. Our previous studies also demonstrated that T1 had potent anti-angiogenesis activity and minimal side effects in vivo. Altogether, this study warrants further investigation to develop T1 as an effective and safe agent for the therapy and prevention of breast cancer.
References
[1]
American Cancer Society (2010) Overview: Breast Cancer. How Many Women Get BreastCancer? Revised: 09/24/2010.
[2]
Lee WY, Chiu LC, Yeung JH (2008) Cytotoxicity of major tanshinones isolated from Danshen (Salvia miltiorrhiza) on HepG2 cells in relation to glutathione perturbation. Food Chem Toxicol 46: 328–338.
[3]
Wang X, Wei Y, Yuan S, Liu G, Lu Y, et al. (2005) Potential anticancer activity of tanshinone IIA against human breast cancer. Int J Cancer 116: 799–807.
[4]
Su CC, Lin YH (2008) Tanshinone IIA inhibits human breast cancer cells through increased Bax to Bcl-xL ratios. Int J Mol Med 22: 357–361.
[5]
Yuan S, Wang Y, Chen X, Song Y, Yang Y (2002) A study on apoptosis of nasopharyngeal carcinoma cell line induced by Tanshinone II A and its molecular mechanism. Hua Xi Yi Ke Da Xue Xue Bao 33: 84–86, 90:
[6]
Wang J, Wang X, Jiang S, Yuan S, Lin P, et al. (2007) Growth inhibition and induction of apoptosis and differentiation of tanshinone IIA in human glioma cells. J Neurooncol 82: 11–21.
[7]
Liu JJ, Zhang Y, Lin DJ, Xiao RZ (2009) Tanshinone IIA inhibits leukemia THP-1 cell growth by induction of apoptosis. Oncol Rep 21: 1075–1081.
[8]
Yuan SL, Wei YQ, Wang XJ, Xiao F, Li SF, et al. (2004) Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells. World J Gastroenterol 10: 2024–2028.
[9]
Wang X, Yuan S, Wang C (1996) A preliminary study of the anti-cancer effect of tanshinone on hepatic carcinoma and its mechanism of action in mice. Zhonghua Zhong Liu Za Zhi 18: 412–414.
[10]
Zhang P, Pei Y, Qi Y (1999) Influence of blood-activating drugs on adhesion and invasion of cells in lung cancer patients. Zhongguo Zhong Xi Yi Jie He Za Zhi 19: 103–105.
[11]
Mosaddik MA (2003) In vitro cytotoxicity of tanshinones isolated from Salvia miltiorrhiza Bunge against P388 lymphocytic leukemia cells. Phytomedicine 10: 682–685.
[12]
Lee CY, Sher HF, Chen HW, Liu CC, Chen CH, et al. (2008) Anticancer effects of tanshinone I in human non-small cell lung cancer. Mol Cancer Ther 7: 3527–3538.
[13]
Nizamutdinova IT, Lee GW, Son KH, Jeon SJ, Kang SS, et al. (2008) Tanshinone I effectively induces apoptosis in estrogen receptor-positive (MCF-7) and estrogen receptor-negative (MDA-MB-231) breast cancer cells. Int J Oncol 33: 485–491.
[14]
Nizamutdinova IT, Lee GW, Lee JS, Cho MK, Son KH, et al. (2008) Tanshinone I suppresses growth and invasion of human breast cancer cells, MDA-MB-231, through regulation of adhesion molecules. Carcinogenesis 29: 1885–1892.
[15]
Meraldi P, Honda R, Nigg EA (2004) Aurora kinases link chromosome segregation and cell division to cancer susceptibility. Curr Opin Genet Dev 14: 29–36.
[16]
Giet R, Prigent C (1999) Aurora/Ipl1p-related kinases, a new oncogenic family of mitotic serine-threonine kinases. J Cell Sci 112 ( Pt 21): 3591–3601.
[17]
Kaestner P, Stolz A, Bastians H (2009) Determinants for the efficiency of anticancer drugs targeting either Aurora-A or Aurora-B kinases in human colon carcinoma cells. Mol Cancer Ther 8: 2046–2056.
[18]
Vankayalapati H, Bearss DJ, Saldanha JW, Munoz RM, Rojanala S, et al. (2003) Targeting aurora2 kinase in oncogenesis: a structural bioinformatics approach to target validation and rational drug design. Mol Cancer Ther 2: 283–294.
[19]
Marumoto T, Zhang D, Saya H (2005) Aurora-A - a guardian of poles. Nat Rev Cancer 5: 42–50.
[20]
Comperat E, Camparo P, Haus R, Chartier-Kastler E, Radenen B, et al. (2007) Aurora-A/STK-15 is a predictive factor for recurrent behaviour in non-invasive bladder carcinoma: a study of 128 cases of non-invasive neoplasms. Virchows Arch 450: 419–424.
[21]
Comperat E, Bieche I, Dargere D, Laurendeau I, Vieillefond A, et al. (2008) Gene expression study of Aurora-A reveals implication during bladder carcinogenesis and increasing values in invasive urothelial cancer. Urology 72: 873–877.
[22]
Staff S, Isola J, Jumppanen M, Tanner M (2010) Aurora-A gene is frequently amplified in basal-like breast cancer. Oncology reports 23: 307–312.
[23]
Lentini L, Amato A, Schillaci T, Insalaco L, Di Leonardo A (2008) Aurora-A transcriptional silencing and vincristine treatment show a synergistic effect in human tumor cells. Oncol Res 17: 115–125.
[24]
Li D, Zhu J, Firozi PF, Abbruzzese JL, Evans DB, et al. (2003) Overexpression of oncogenic STK15/BTAK/Aurora A kinase in human pancreatic cancer. Clin Cancer Res 9: 991–997.
[25]
Lee EC, Frolov A, Li R, Ayala G, Greenberg NM (2006) Targeting Aurora kinases for the treatment of prostate cancer. Cancer Res 66: 4996–5002.
[26]
Matarasso N, Bar-Shira A, Rozovski U, Rosner S, Orr-Urtreger A (2007) Functional analysis of the Aurora Kinase A Ile31 allelic variant in human prostate. Neoplasia 9: 707–715.
[27]
Qu Y, Zhang L, Mao M, Zhao F, Huang X, et al. (2008) Effects of DNAzymes targeting Aurora kinase A on the growth of human prostate cancer. Cancer Gene Ther 15: 517–525.
[28]
Qu Y, Zhang L, Mu DZ, Huang X, Wei DP, et al. (2008) Effect of aurora A on carcinogenesis of human prostate cancer. Sichuan Da Xue Xue Bao Yi Xue Ban 39: 1–5.
[29]
Buschhorn HM, Klein RR, Chambers SM, Hardy MC, Green S, et al. (2005) Aurora-A over-expression in high-grade PIN lesions and prostate cancer. Prostate 64: 341–346.
[30]
Keen N, Taylor S (2004) Aurora-kinase inhibitors as anticancer agents. Nat Rev Cancer 4: 927–936.
[31]
Dar AA, Goff LW, Majid S, Berlin J, El-Rifai W (2010) Aurora kinase inhibitors--rising stars in cancer therapeutics? Mol Cancer Ther 9: 268–278.
[32]
Gorgun G, Calabrese E, Hideshima T, Ecsedy J, Perrone G, et al. (2010) A novel aurora-A kinase inhibitor MLN8237 induces cytotoxicity and cell cycle arrest in multiple myeloma. Blood 115: 5202–5213.
[33]
Gong Y, Li Y, Lu Y, Li L, Abdolmaleky HM, et al. (2011) Bioactive tanshinones in Salvia Miltiorrhiza inhibit the growth of prostate cancer cells in vitro and in mice. Int J Cancer 129: 1042–1052.
[34]
Lima SC, Hernandez-Vargas H, Herceg Z (2010) Epigenetic signatures in cancer: Implications for the control of cancer in the clinic. Current opinion in molecular therapeutics 12: 316–324.
[35]
Banerjee HN, Verma M (2009) Epigenetic mechanisms in cancer. Biomarkers in medicine 3: 397–410.
[36]
Dworkin AM, Huang TH, Toland AE (2009) Epigenetic alterations in the breast: Implications for breast cancer detection, prognosis and treatment. Seminars in cancer biology 19: 165–171.
[37]
Veeck J, Esteller M (2010) Breast cancer epigenetics: from DNA methylation to microRNAs. Journal of mammary gland biology and neoplasia 15: 5–17.
[38]
Davie JR (2003) Inhibition of histone deacetylase activity by butyrate. The Journal of nutrition 133: 2485S–2493S.
[39]
Lea MA, Rasheed M, Randolph VM, Khan F, Shareef A, et al. (2002) Induction of histone acetylation and inhibition of growth of mouse erythroleukemia cells by S-allylmercaptocysteine. Nutrition and cancer 43: 90–102.
[40]
Fang M, Chen D, Yang CS (2007) Dietary polyphenols may affect DNA methylation. The Journal of nutrition 137: 223S–228S.
[41]
Cui Y, Lu C, Liu L, Sun D, Yao N, et al. (2008) Reactivation of methylation-silenced tumor suppressor gene p16INK4a by nordihydroguaiaretic acid and its implication in G1 cell cycle arrest. Life sciences 82: 247–255.
[42]
Bi HC, Law FC, Zhong GP, Xu CS, Pan Y, et al. (2007) Study of tanshinone IIA tissue distribution in rat by liquid chromatography-tandem mass spectrometry method. Biomed Chromatogr 21: 473–479.
[43]
Song M, Hang TJ, Zhang Zh X, Du R, Chen J (2005) Determination of cryptotanshinone and its metabolite in rat plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 827: 205–209.
[44]
Hao H, Wang G, Li P, Li J, Ding Z (2006) Simultaneous quantification of cryptotanshinone and its active metabolite tanshinone IIA in plasma by liquid chromatography/tandem mass spectrometry (LC-MS/MS). J Pharm Biomed Anal 40: 382–388.
[45]
Li J, Wang G, Li P, Hao H (2005) Simultaneous determination of tanshinone IIA and cryptotanshinone in rat plasma by liquid chromatography-electrospray ionisation-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 826: 26–30.
[46]
Li Y, Gong Y, Li L, Abdolmaleky HM, Zhou J-R (2012) Bioactive tanshinone I inhibits the growth of lung cancer in part via downregulation of Aurora A function. Molecular Carcinogenesis in press.
[47]
Singh AV, Franke AA, Blackburn GL, Zhou JR (2006) Soy phytochemicals prevent orthotopic growth and metastasis of bladder cancer in mice by alterations of cancer cell proliferation and apoptosis and tumor angiogenesis. Cancer Res 66: 1851–1858.
[48]
Abdolmaleky HM, Smith CL, Zhou J-R, Thiagalingam S (2008) Epigenetic alterations of the dopaminergic system in major psychiatric disorders. Methods Mol Biol 448: 187–212.
[49]
Mai Z, Blackburn GL, Zhou JR (2007) Soy phytochemicals synergistically enhance the preventive effect of tamoxifen on the growth of estrogen-dependent human breast carcinoma in mice. Carcinogenesis 28: 1217–1223.
