Growth Inhibition and Apoptosis Induction by Tanshinone IIA in Human Colon Adenocarcinoma Cells

Abstract

Tanshinone IIA is the most abundant diterpene quinone in Danshen, Salviae miltiorrhizae Radix, a widely prescribed traditional herbal medicine that is used to treat cardiovascular and inflammatory diseases. Recently, tanshinone IIA was demonstrated to induce cell death and apoptosis in a variety of tumors. However, the effect of tanshinone IIA on human colon cancer cells is not clearly understood yet. In this study, the antigrowth and apoptosis-eliciting effects of tanshinone IIA, as well as its cellular mechanisms of actions, were investigated in Colo-205 human colon cancer cells. Tanshinone IIA reduced cell growth in a concentration-dependent manner, inducing apoptosis accompanied by an increase in TUNEL staining and by an increased percentage of cells in the sub-G1 fraction. The expression of p53 and p21 and mitochondrial cytochrome c release were increased in tanshinone IIA-treated cells. In addition, the expression of Fas proteins was up-regulated by tanshinone IIA. Tanshinone IIA-induced catalytic activation of caspases was confirmed by cleavage of caspase-8 and caspase-3. These findings suggest that tanshinone IIA induces apoptosis in Colo-205 cells through both mitochondrial-mediated intrinsic and Fas-mediated extrinsic caspase cell-death pathways. Accordingly, the chemotherapeutic potential of tanshinone IIA for colon cancer warrants further study.

tanshinone IIA - apoptosis - Colo-205 - Fas - caspase

References

• 1 Jemal A, Siegel R, Ward E, Murray T, Xu J, Smigal C. et al . Cancer statistics, 2006.  CA Cancer J Clin. 2006;  56 106-30
• 2 Verhoef M J, Balneaves L G, Boon H S, Vroegindewey A. Reasons for and characteristics associated with complementary and alternative medicine use among adult cancer patients: a systematic review.  Integr Cancer Ther. 2005;  4 274-86
• 3 Boon H, Wong J. Botanical medicine and cancer: a review of the safety and efficacy.  Expert Opin Pharmacother. 2004;  5 2485-501
• 4 Fish J M, Welchons D R, Kim Y S, Lee S H, Ho W K, Antzelevitch C. Dimethyl lithospermate B, an extract of Danshen, suppresses arrhythmogenesis associated with the Brugada syndrome.  Circulation. 2006;  113 1393-400
• 5 Chang P N, Mao J C, Huang S H, Ning L, Wang Z J, On T. et al . Analysis of cardioprotective effects using purified Salvia miltiorrhiza extract on isolated rat hearts.  J Pharmacol Sci. 2006;  101 245-9
• 6 Che A J, Zhang J Y, Li C H, Chen X F, Hu Z D, Chen X G. Separation and determination of active components in Radix Salviae miltiorrhizae and its medicinal preparations by nonaqueous capillary electrophoresis.  J Sep Sci. 2004;  27 569-75
• 7 Zhou L, Zuo Z, Chow M S. Danshen: an overview of its chemistry, pharmacology, pharmacokinetics, and clinical use.  J Clin Pharmacol. 2005;  45 1345-59
• 8 Lin R, Wang W R, Liu J T, Yang G D, Han C J. Protective effect of tanshinone IIA on human umbilical vein endothelial cell injured by hydrogen peroxide and its mechanism.  J Ethnopharmacol. 2006;  108 217-22
• 9 Wang A M, Sha S H, Lesniak W, Schacht J. Tanshinone (Salviae miltiorrhizae extract) preparations attenuate aminoglycoside-induced free radical formation in vitro and ototoxicity in vivo.  Antimicrob Agents Chemother. 2003;  47 1836-41
• 10 Jang S I, Kim H J, Kim Y J, Jeong S I, You Y O. Tanshinone IIA inhibits LPS-induced NF-kappaB activation in RAW 264.7 cells: possible involvement of the NIK-IKK, ERK1/2, p38 and JNK pathways.  Eur J Pharmacol. 2006;  542 1-7
• 11 Li W, Li J, Ashok M, Wu R, Chen D, Yang L. et al . A cardiovascular drug rescues mice from lethal sepsis by selectively attenuating a late-acting proinflammatory mediator, high mobility group box 1.  J Immunol. 2007;  78 3856-64
• 12 Ryu S Y, Lee C O, Choi S U. In vitro cytotoxicity of tanshinones from Salvia miltiorrhiza.  Planta Med. 1997;  63 339-42
• 13 Wang X, Morris-Natschke S L, Lee K H. New developments in the chemistry and biology of the bioactive constituents of Tanshen.  Med Res Rev. 2007;  27 133-48
• 14 Sung H J, Choi S M, Yoon Y, An K S. Tanshinone IIA, an ingredient of Salvia miltiorrhiza Bunge, induces apoptosis in human leukemia cell lines through the activation of caspase-3.  Exp Mol Med. 1999;  31 174-8
• 15 Liu J J, Lin D J, Liu P Q, Huang M, Li X D, Huang R W. Induction of apoptosis and inhibition of cell adhesive and invasive effects by tanshinone IIA in acute promyelocytic leukemia cells in vitro.  J Biomed Sci. 2006;  13 813-23
• 16 Wang X, Wei Y, Yuan S, Liu G, Lu Y, Zhang J. et al . Potential anticancer activity of tanshinone IIA against human breast cancer.  Int J Cancer. 2005;  116 799-807
• 17 Yuan S L, Wei Y Q, Wang X J, Xiao F, Li S F, Zhang J. Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells.  World J Gastroenterol. 2004;  10 2024-8
• 18 Li T M, Chen G W, Su C C, Lin J G, Yeh C C, Cheng K C. et al . Ellagic acid induced p53/p21 expression, G1 arrest and apoptosis in human bladder cancer T24 cells.  Anticancer Res. 2005;  25 971-9
• 19 Chen H C, Hsieh W T, Chang W C, Chung J G. Aloe-emodin induced in vitro G2/M arrest of cell cycle in human promyelocytic leukemia HL-60 cells.  Food Chem Toxicol. 2004;  42 1251-7
• 20 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Anal Biochem. 1976;  72 248-54
• 21 Gartel A L, Tyner A L. The role of the cyclin-dependent kinase inhibitor p21 in apoptosis.  Mol Cancer Ther. 2002;  1 639-49
• 22 Yuan S, Wang Y, Chen X, Song Y, Yang Y. 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. 2002;  33 84-6
• 23 Broker L E, Kruyt F A, Giaccone G. Cell death independent of caspases: a review.  Clin Cancer Res. 2005;  11 3155-62
• 24 Mihara M, Erster S, Zaika A, Petrenko O, Chittenden T, Pancoska P. et al . p53 has a direct apoptogenic role at the mitochondria.  Mol Cell. 2003;  11 577-90
• 25 Cohen G M. Caspases: the executioners of apoptosis.  Biochem J. 1997;  326 1-16
• 26 Lowe S W, Cepero E, Evan G. Intrinsic tumour suppression.  Nature. 2004;  432 307-15
• 27 Scaffidi C, Fulda S, Srinivasan A, Friesen C, Li F, Tomaselli K J. et al . Two CD95 (APO-1/Fas) signaling pathways.  Embo J. 1998;  17 1675-87
• 28 Tamura T, Aoyama N, Saya H, Haga H, Futami S, Miyamoto M. et al . Induction of Fas-mediated apoptosis in p53-transfected human colon carcinoma cells.  Oncogene. 1995;  11 1939-46
• 29 Petak I, Tillman D M, Houghton J A. p53 dependence of Fas induction and acute apoptosis in response to 5-fluorouracil-leucovorin in human colon carcinoma cell lines.  Clin Cancer Res. 2000;  6 4432-41