Tanshinone II-A is an alcohol extract of the root of the traditional Chinese medicinal plant Salvia miltiorrhiza Bunge, whose effects and mechanism in tumor metastasis are still unclear. The aim of this study was to investigate the effects of tanshinone II-A on tumor invasion and metastasis in human hepatocellular carcinoma (HCC) and its possible mechanism of action.
Methods and Study Design
The HCC cell lines HepG2 and SMMC-7721 were treated with tanshinone II-A at different doses. Invasion and metastasis of tumor cells were examined by in vitro and in vivo assays. The molecular mechanisms of tanshinone II-A for inhibiting invasion and metastasis of HCC cells were investigated by Western blot and gelatin zymography.
Results
Treatment with tanshinone II-A had inhibitory effects on the migration and invasion of HCC cells. Increasing doses resulted in enhanced inhibitory effects. At 0.5 mg/L, the inhibitory effect was noticeable. At 1 mg/L, the inhibitory rate was 53.15%. The inhibitory effect became stronger with time; among 24, 48, 72 and 96 hours of treatment, the most significant effects were observed at 72 hours. Tanshinone II-A also significantly inhibited in vivo metastasis of HepG2 cells. Tanshinone II-A inhibited in vitro and in vivo invasion and metastasis of HCC cells by reducing the expression of the metalloproteinases MMP2 and MMP9 and by blocking NF-kappa B activation.
Conclusions
Tanshinone II-A effectively inhibited invasion and metastasis of HCC cells in vitro and in vivo, partly by inhibiting the activity of MMP2 and MMP9, and partly via the NF-kappa B signal transduction pathway.
ParkinD.M., BrayF., FerlayJ.: Global cancer statistics, 2000.CA Cancer J Clin, 55: 74–108, 2002.
2.
YuanS.L., WeiY.Q., WangX.J., XiaoF., LiS.F., ZhangJ.: Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells.World J Gastroenterol, 10: 2024–2028, 2004.
3.
El-SeragH.B., MasonA.C.: Rising incidence of hepatocellular carcinoma in the United States.N Engl J Med, 340: 745–750, 1999.
4.
De Vos IrvineH., GoldbergD., HoleD.J., McMenaminJ.: Trends in primary liver cancer.Lancet, 351: 215–216, 1998.
5.
ZhangQ., KangX., ZhaoW.: Antiangiogenic effect of low-dose cyclophosphamide combined with ginsenoside Rg3 on Lewis lung carcinoma.Biochem Biophys Res Commun, 342: 824–828, 2006.
6.
KangX.M., ZhangQ.Y., TongD.D., ZhaoW.: Experimental study on anti-angiogenesis in mice with Lewis lung carcinoma by low-dose of cyclophosphamide combined with ginsenoside Rg3.Zhongguo Zhong Xi Yi Jie He Za Zhi, 25: 730–733, 2005.
7.
IishiH., TatsutaM., BabaM., UeharaH., NakaizumiA., ShinkaiK., AkedoH., FunaiH., IshiguroS., KitagawaI.: Inhibition by ginsenoside Rg3 of bombesin-enhanced peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane in Wistar rats.Clin Exp Metastasis, 15: 603–611, 1997.
8.
ChenJ., PengH., Ou-YangX., HeX.: Research on the antitumor effect of ginsenoside Rg3 in B16 melanoma cells.Melanoma Res, 18: 322–329, 2008.
9.
KimH.S., LeeE.H., KoS.R., ChoiK.J., ParkJ.H., ImD.S.: Effects of ginsenosides Rg3 and Rh2 on the proliferation of prostate cancer cells.Arch Pharm Res, 27: 429–435, 2004.
10.
LiuW.K., XuS.X., CheC.T.: Anti-proliferative effect of ginseng saponins on human prostate cancer cell line.Life Sci, 67: 1297–1306, 2000.
11.
YuanS.L., HuangR.M., WangX.J., SongY., HuangG.Q.: Reversing effect of Tanshinone on malignant phenotypes of human hepatocarcinoma cell line.World J Gastroenterol, 4: 317–319, 1998.
12.
YuanS.L., WeiY.Q., WangX.J., XiaoF., LiS.F., ZhangJ.: Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells.World J Gastroenterol, 10: 2024–2028, 2004.
13.
WuW. L., ChangW.L., ChenC.F.: Cytotoxic activities of tanshinones against human carcinoma cell lines.Am J Chin Med, 19: 207–216, 1991.
14.
YoonY., KimY.O., JeonW.K., ParkH.J., SungH.J.: Tanshinone IIA isolated from Salvia miltiorrhiza BUNGE induced apoptosis in HL60 human premyelocytic leukemia cell line.J Ethnopharmacol, 68: 121–127, 1999.
15.
SungH.J., ChoiS.M., YoonY., AnK.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, 31: 174–178, 1999.
16.
AiZ.W.: Reversing effect of retinoic acid on some phenotypes of human hepatocarcinoma cell line.Zhonghua Zhongliu Zazhi, 13: 9–12, 1991.
17.
HaifengJ., YanglinP., LijieH., HuihongZ., XiaohuaL., LinaZ., LiS., JieL., LiuH., JiugangS., HuahongX., JuanG., ShuangH., YingL., DaimingF.: p75 neurotrophin receptor inhibits invasion and metastasis of gastric cancer.Mol Cancer Res, 5: 423–433, 2007.
18.
SunF.X., TangZ.Y., LuiK.D., YeS.L., XueQ., GaoD.M., MaZ.C.: Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues.Int J Cancer, 66: 239–243, 1996.
19.
TianJ., TangZ.Y., YeS.L., LiuY.K., LinZ.Y., ChenJ., XueQ.: New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis.Br J Cancer, 81: 814–821, 1999.
20.
KondoS., KubotaS., ShimoT., NishidaT., YosimichiG., EguchiT., SugaharaT., TakigawaM.: Connective tissue growth factor increased by hypoxia may initiate angiogenesis in collaboration with matrix metalloproteinases.Carcinogenesis, 23: 769–776, 2002.
21.
PeiL., ZhenghuaiT., ZhifengZ., HonghaoL., ZhengjiM.: Inhibitory effects of salvianolic acid B on the high glucose-induced mesangial proliferation via NF-κ B-dependent pathway.Biol Pharm Bull, 31: 1381–1386, 2008.
AuwardtR.B., MudgeS.J., ChenC., PowerD.A.: Inhibition with antisense oligonucleotide suggests that IkappaB-alpha does not form a negative autoregulatory loop for NF-kappaB in mesangial cells.Exp Nephrol, 8: 144–151, 2000.
25.
ParkinD.M.: Global cancer statistics in the year 2000.Lancet Oncol, 2: 533–543, 2001.
26.
TangZ.Y., YeS.L., LiuY.K., QinL.X., SunH.C., YeQ.H., WangL., ZhouJ., QiuS.J., LiY., JiX.N., LiuH., XiaJ.L., WuZ.Q., FanJ., MaZ.C., ZhouX.D., LinZ.Y., LiuK.D.: A decade's studies on metastasis of hepatocellular carcinoma.J Cancer Res Clin Oncol, 130: 187–196, 2004.
27.
ChoJ.Y., ChungH.C., NohS.H., RohJ.K., MinJ.S., KimB.S.: High level of urokinase-type plasminogen activator is a new prognostic marker inpatients with gastric carcinoma.Cancer, 79: 878–883, 1997.
28.
IwamotoJ., MizokamiY., TakahashiK., NakajimaK., OhtsuboT., MiuraS., NarasakaT., TakeyamaH., OmataT., ShimokobeK., ItoM., TakeharaH., MatsuokaT.: Expressions of urokinase-type plasminogen activator, its receptor and plasminogen activator inhibitor-1 in gastric cancer cells and effects of Helicobacter pylori.Scand J Gastroenterol, 40: 783–793, 2005.
29.
FarinaA.R., CoppaA., TiberioA., TacconelliA., TurcoA., CollettaG., GulinoA., MackayA.R.: Transforming growth factorbeta1 enhances the invasiveness of human MDA-MB-231 breast cancer cells by up-regulating urokinase activity.Int J Cancer, 75: 721–730, 1998.
30.
KanekoT., KonnoH., BabaM., TanakaT., NakamuraS.: Urokinase-type plasminogen activator expression correlates with tumor angiogenesis and poor outcome in gastric cancer.Cancer Sci, 94: 43–49, 2003.
31.
MatsumuraS., OueN., NakayamaH., KitadaiY., YoshidaK., YamaguchiY., ImaiK., NakachiK., MatsusakiK., ChayamaK., YasuiW.: A single nucleotide polymorphism in the MMP-9 promoter affects tumor progression and invasive phenotype of gastric cancer.J Cancer Res Clin Oncol, 131: 19–25, 2005.
32.
AlfanoD., FrancoP., VoccaI., GambiN., PisaV., ManciniA., CaputiM., CarrieroM.V., IaccarinoI., StoppelliM.P.: The urokinase plasminogen activator and its receptor: role in cell growth and apoptosis.Thromb Haemost, 93: 205–211, 2005.
33.
JinU.H., SuhS.J., ChangH.W., SonJ.K., LeeS.H., SonK.H., ChangY.C., KimC.H.: Tanshinone IIA from Salvia miltiorrhiza BUNGE inhibits human aortic smooth muscle cell migration and MMP-9 activity through AKT signaling pathway.J Cell Biochem, 104: 15–26, 2008.
34.
BoursV., BonizziG., Bentires-AljM., BureauF., PietteJ., LekeuxP., MervilleM.: NF-kappaB activation in response to toxical and therapeutical agents: role in inflammation and cancer treatment.Toxicology, 153: 27–38, 2000.
35.
HenkelT., MachleidtT., AlkalayI., KronkeM., Ben-NeriahY., BaeuerleP.A.: Rapid proteolysis of I kappa B-alpha is necessary for activation of transcription factor NF-kappa B.Nature, 365: 182–185, 1993.
