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01-01-2012 | Original Paper

Tanshinone IIA activates calcium-dependent apoptosis signaling pathway in human hepatoma cells

Authors: Zhi-Kai Dai, Jiang-Ke Qin, Jiao-E Huang, Yong Luo, Qing Xu, Hai-Lu Zhao

Published in: Journal of Natural Medicines | Issue 1/2012

Abstract

Tanshinone IIA (Tan IIA), a natural product from herb Salvia miltiorrhiza Bunge, has potential anti-tumor activity. The aim of this study was to pinpoint the molecular mechanisms underlying Tan IIA-induced cancer cell apoptosis. Human hepatoma BEL-7402 cells treated with Tan IIA underwent assessment with MTT assay for cell viability, 10-day culture for colony formation, flow cytometry and fluorescence microscopy for apoptosis and cell cycle analysis. Changes in intracellular [Ca²⁺] and mitochondrial membrane potential (∆ψ) reflected the calcium-dependent apoptosis pathway. RT-PCR was used to detect gene expression of Bad and metallothionein 1A (MT 1A). Cytotoxicity of Tan IIA was tested in human amniotic mesenchymal stem cells (HAMCs). Tan IIA exhibited dose-dependent and time-dependent anticancer effects on BEL-7402 cells through apoptosis and G₀/G₁ arrest. Cells treated with Tan IIA increased their intracellular calcium, decreased their mitochondrial membrane potential and induced Bad and MT 1A mRNA expression. No adverse effects of Tan IIA were found in HAMCs. In conclusion, these results indicate that Tan IIA-induced cancer cell apoptosis acts via activation of calcium-dependent apoptosis signaling pathways and upregulation of MT 1A expression.

Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM (2010) Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 127:2893–2917PubMedCrossRef

Farazi PA, DePinho RA (2006) Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer 6:674–687PubMedCrossRef

Zhou L, Zuo Z, Chow MS (2005) Danshen: an overview of its chemistry, pharmacology, pharmacokinetics, and clinical use. J Clin Pharmacol 45:1345–1359PubMedCrossRef

Niu XL, Ichimori K, Yang X, Hirota Y, Hoshiai K, Li M, Nakazawa H (2000) Tanshinone II-A inhibits low density lipoprotein oxidation in vitro. Free Radic Res 33:305–312PubMedCrossRef

Yan FF, Liu YF, Liu Y, Zhao YX (2009) Sulfotanshinone Sodium Injection could decrease fibrinogen level and improve clinical outcomes in patients with unstable angina pectoris. Int J Cardiol 135:254–255PubMedCrossRef

Dong X, Dong J, Zhang R, Fan L, Liu L, Wu G (2009) Anti-inflammatory effects of tanshinone IIA on radiation-induced microglia BV-2 cells inflammatory response. Cancer Biother Radiopharm 24:681–687PubMedCrossRef

Kapoor S (2009) Tanshinone IIA: a potent, natural anti-carcinogenic agent for the management of systemic malignancies. Chin J Integr Med 15:153PubMedCrossRef

Yoon Y, Kim YO, Jeon WK, Park HJ, Sung HJ (1999) Tanshinone IIA isolated from Salvia miltiorrhiza BUNGE induced apoptosis in HL60 human premyelocytic leukemia cell line. J Ethnopharmacol 68:121–127PubMedCrossRef

Lee WY, Cheung CC, Liu KW, Fung KP, Wong J, Lai PB, Yeung JH (2010) Cytotoxic effects of tanshinones from Salvia miltiorrhiza on doxorubicin-resistant human liver cancer cells. J Nat Prod 73:854–859PubMedCrossRef

10.

Tang Z, Tang Y, Fu L (2003) Growth inhibition and apoptosis induction in human hepatoma cells by tanshinone II A. J Huazhong Univ Sci Technol Med Sci 23:166–168, 172

11.

Zhu B, Zhai Q, Yu B (2010) Tanshinone IIA protects rat primary hepatocytes against carbon tetrachloride toxicity via inhibiting mitochondria permeability transition. Pharm Biol 48:484–487PubMedCrossRef

12.

Shi SL, Li QF, Liu QR, Xu DH, Tang J, Liang Y, Zhao ZL, Yang LM (2009) Nuclear matrix protein, prohibitin, was down-regulated and translocated from nucleus to cytoplasm during the differentiation of osteosarcoma MG-63 cells induced by ginsenoside Rg1, cinnamic acid, and tanshinone IIA (RCT). J Cell Biochem 108:926–934PubMedCrossRef

13.

Yeruva L, Hall C, Elegbede JA, Carper SW (2010) Perillyl alcohol and methyl jasmonate sensitize cancer cells to cisplatin. Anticancer Drugs 21:1–9PubMedCrossRef

14.

Ward MW, Rego AC, Frenguelli BG, Nicholls DG (2000) Mitochondrial membrane potential and glutamate excitotoxicity in cultured cerebellar granule cells. J Neurosci 20:7208–7219PubMed

15.

Ni X, Sullivan GJ, Martin-Caraballo M (2007) Developmental characteristics of AMPA receptors in chick lumbar motoneurons. Dev Neurobiol 67:1419–1432PubMedCrossRef

16.

Miao Z, Jin J, Chen L, Zhu J, Huang W, Zhao J, Qian H, Zhang X (2006) Isolation of mesenchymal stem cells from human placenta: comparison with human bone marrow mesenchymal stem cells. Cell Biol Int 30:681–687PubMedCrossRef

17.

Mizokami T, Hisha H, Okazaki S, Takaki T, Wang XL, Song CY, Li Q, Kato J, Hosaka N, Inaba M, Kanzaki H, Ikehara S (2009) Preferential expansion of human umbilical cord blood-derived CD34-positive cells on major histocompatibility complex-matched amnion-derived mesenchymal stem cells. Haematologica 94:618–628PubMedCrossRef

18.

Yuan SL, Wei YQ, Wang XJ, Xiao F, Li SF, Zhang J (2004) Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells. World J Gastroenterol 10:2024–2028PubMed

19.

Wang ZY (2003) Ham-Wasserman lecture: treatment of acute leukemia by inducing differentiation and apoptosis. Hematology Am Soc Hematol Educ Program 1–13

20.

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 18:412–414PubMed

21.

Soleimanpour SA, Crutchlow MF, Ferrari AM, Raum JC, Groff DN, Rankin MM, Liu C, De Leon DD, Naji A, Kushner JA, Stoffers DA (2010) Calcineurin signaling regulates human islet {beta}-cell survival. J Biol Chem 285:40050–40059PubMedCrossRef

22.

Wang X, Wei Y, Yuan S, Liu G, Lu Y, Zhang J, Wang W (2005) Potential anticancer activity of tanshinone IIA against human breast cancer. Int J Cancer 116:799–807PubMedCrossRef

23.

Liu Y, Chen H, Jiang Y (2001) Protective effect of tanshinone IIA on acute hepatic injury in mice. Zhong Yao Cai 24:588–589PubMed

24.

Liu Y, Wang X (2003) Protective effects of tanshinone IIA on injured primary cultured rat hepatocytes induced by CCl4. Zhong Yao Cai 26:415–417PubMed

25.

Lu Q, Zhang P, Zhang X, Chen J (2009) Experimental study of the anti-cancer mechanism of tanshinone IIA against human breast cancer. Int J Mol Med 24:773–780PubMedCrossRef

26.

Olofsson MH, Havelka AM, Brnjic S, Shoshan MC, Linder S (2008) Charting calcium-regulated apoptosis pathways using chemical biology: role of calmodulin kinase II. BMC Chem Biol 8:2PubMedCrossRef

27.

Williams JP, Micoli K, McDonald JM (2010) Calmodulin - an often-ignored signal in osteoclasts. Ann N Y Acad Sci 1192:358–364PubMedCrossRef

28.

Devireddy LR, Green MR (2003) Transcriptional program of apoptosis induction following interleukin 2 deprivation: identification of RC3, a calcium/calmodulin binding protein, as a novel proapoptotic factor. Mol Cell Biol 23:4532–4541PubMedCrossRef

29.

Crabtree GR (2001) Calcium, calcineurin, and the control of transcription. J Biol Chem 276:2313–2316PubMedCrossRef

30.

Shibasaki F, McKeon F (1995) Calcineurin functions in Ca(2+)-activated cell death in mammalian cells. J Cell Biol 131:735–743PubMedCrossRef

31.

Wang HG, Pathan N, Ethell IM, Krajewski S, Yamaguchi Y, Shibasaki F, McKeon F, Bobo T, Franke TF, Reed JC (1999) Ca2+-induced apoptosis through calcineurin dephosphorylation of BAD. Science 284:339–343PubMedCrossRef

32.

Willis SN, Chen L, Dewson G, Wei A, Naik E, Fletcher JI, Adams JM, Huang DC (2005) Proapoptotic Bak is sequestered by Mcl-1 and Bcl-xL, but not Bcl-2, until displaced by BH3-only proteins. Genes Dev 19:1294–1305PubMedCrossRef

33.

Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB, Korsmeyer SJ (2001) Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292:727–730PubMedCrossRef

34.

Shimizu S, Narita M, Tsujimoto Y (1999) Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399:483–487PubMedCrossRef

35.

Sullivan PG, Dragicevic NB, Deng JH, Bai Y, Dimayuga E, Ding Q, Chen Q, Bruce-Keller AJ, Keller JN (2004) Proteasome inhibition alters neural mitochondrial homeostasis and mitochondria turnover. J Biol Chem 279:20699–20707PubMedCrossRef

36.

Rivera A, Maxwell SA (2005) The p53-induced gene-6 (proline oxidase) mediates apoptosis through a calcineurin-dependent pathway. J Biol Chem 280:29346–29354PubMedCrossRef

37.

Chiu TL, Su CC (2010) Tanshinone IIA induces apoptosis in human lung cancer A549 cells through the induction of reactive oxygen species and decreasing the mitochondrial membrane potential. Int J Mol Med 25:231–236PubMed

38.

Sastry KS, Smith AJ, Karpova Y, Datta SR, Kulik G (2006) Diverse antiapoptotic signaling pathways activated by vasoactive intestinal polypeptide, epidermal growth factor, and phosphatidylinositol 3-kinase in prostate cancer cells converge on BAD. J Biol Chem 281:20891–20901PubMedCrossRef

39.

She QB, Solit DB, Ye Q, O’Reilly KE, Lobo J, Rosen N (2005) The BAD protein integrates survival signaling by EGFR/MAPK and PI3 K/Akt kinase pathways in PTEN-deficient tumor cells. Cancer Cell 8:287–297PubMedCrossRef

40.

Theocharis SE, Margeli AP, Klijanienko JT, Kouraklis GP (2004) Metallothionein expression in human neoplasia. Histopathology 45:103–118PubMedCrossRef

41.

Alonso-Gonzalez C, Mediavilla D, Martinez-Campa C, Gonzalez A, Cos S, Sanchez-Barcelo EJ (2008) Melatonin modulates the cadmium-induced expression of MT-2 and MT-1 metallothioneins in three lines of human tumor cells (MCF-7, MDA-MB-231 and HeLa). Toxicol Lett 181:190–195PubMedCrossRef

42.

Saito T, Tezuka T, Konno R, Fujii N (2010) Protective effects of metallothionein I and II against metal- and ultraviolet radiation-induced damage in cultured lens epithelial cells. Jpn J Ophthalmol 54:486–493PubMedCrossRef

43.

Majumder S, Roy S, Kaffenberger T, Wang B, Costinean S, Frankel W, Bratasz A, Kuppusamy P, Hai T, Ghoshal K, Jacob ST (2010) Loss of metallothionein predisposes mice to diethylnitrosamine-induced hepatocarcinogenesis by activating NF-kappaB target genes. Cancer Res 70:10265–10276PubMedCrossRef

44.

de Gruijl FR (2000) Photocarcinogenesis: UVA vs UVB. Methods Enzymol 319:359–366PubMedCrossRef

45.

Ioachim E, Assimakopoulos D, Peschos D, Zissi A, Skevas A, Agnantis NJ (1999) Immunohistochemical expression of metallothionein in benign premalignant and malignant epithelium of the larynx: correlation with p53 and proliferative cell nuclear antigen. Pathol Res Pract 195:809–814PubMedCrossRef

46.

Klaassen CD, Liu J, Choudhuri S (1999) Metallothionein: an intracellular protein to protect against cadmium toxicity. Annu Rev Pharmacol Toxicol 39:267–294PubMedCrossRef

47.

Hu R, Hebbar V, Kim BR, Chen C, Winnik B, Buckley B, Soteropoulos P, Tolias P, Hart RP, Kong AN (2004) In vivo pharmacokinetics and regulation of gene expression profiles by isothiocyanate sulforaphane in the rat. J Pharmacol Exp Ther 310:263–271PubMedCrossRef

48.

Yeh CT, Yen GC (2005) Effect of sulforaphane on metallothionein expression and induction of apoptosis in human hepatoma HepG2 cells. Carcinogenesis 26:2138–2148PubMedCrossRef

49.

Cherian MG, Jayasurya A, Bay BH (2003) Metallothioneins in human tumors and potential roles in carcinogenesis. Mutat Res 533:201–209PubMedCrossRef

50.

Krizkova S, Masarik M, Majzlik P, Kukacka J, Kruseova J, Adam V, Prusa R, Eckschlager T, Stiborova M, Kizek R (2010) Serum metallothionein in newly diagnosed patients with childhood solid tumours. Acta Biochim Pol 57:561–566PubMed

51.

Yap X, Tan HY, Huang J, Lai Y, Yip GW, Tan PH, Bay BH (2009) Over-expression of metallothionein predicts chemoresistance in breast cancer. J Pathol 217:563–570PubMedCrossRef

52.

Jin R, Huang J, Tan PH, Bay BH (2004) Clinicopathological significance of metallothioneins in breast cancer. Pathol Oncol Res 10:74–79PubMedCrossRef

53.

Xia WJ, Yang M, Fok TF, Li K, Chan WY, Ng PC, Ng HK, Chik KW, Wang CC, Gu GJ, Woo KS, Fung KP (2005) Partial neuroprotective effect of pretreatment with tanshinone IIA on neonatal hypoxia-ischemia brain damage. Pediatr Res 58:784–790PubMedCrossRef

Title: Tanshinone IIA activates calcium-dependent apoptosis signaling pathway in human hepatoma cells
Authors: Zhi-Kai Dai
Jiang-Ke Qin
Jiao-E Huang
Yong Luo
Qing Xu
Hai-Lu Zhao
Publication date: 01-01-2012
Publisher: Springer Japan
Published in: Journal of Natural Medicines / Issue 1/2012
Print ISSN: 1340-3443
Electronic ISSN: 1861-0293
DOI: https://doi.org/10.1007/s11418-011-0576-0

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Tanshinone IIA activates calcium-dependent apoptosis signaling pathway in human hepatoma cells

Abstract

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Springer Medicine

Abstract

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