丹参酮Ⅱ_A对乳腺癌细胞阿霉素化疗敏感性的影响及相关机制研究
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摘要
目的探讨丹参酮IIA对乳腺癌细胞阿霉素化疗敏感性的影响及相关机制。方法分别用丹参酮IIA、阿霉素、阿霉素+丹参酮IIA干预人乳腺癌MCF-7和阿霉素耐药的MCF-7(MCF-7/dox)细胞,采用MTS法检测细胞增殖率,流式细胞术检测细胞凋亡率,划痕实验观察细胞迁移能力,Western blotting法检测腺瘤样结肠息肉易感蛋白(APC)、β-catenin、钙黏附蛋白E(E-cadherin)、基质金属蛋白酶-2(MMP-2)、MMP-9蛋白的表达。结果丹参酮IIA能明显增强阿霉素对MCF-7和MCF-7/dox细胞的增殖抑制、迁移抑制和诱导凋亡作用;与阿霉素敏感的MCF-7细胞比较,MCF-7/dox细胞中APC蛋白表达水平显著降低(P<0.05),β-catenin蛋白表达水平显著提高(P<0.05);与单用阿霉素比较,阿霉素与丹参酮IIA联合使用能明显上调MCF-7及MCF-7/dox细胞内APC及E-cadherin表达水平(P<0.05),下调β-catenin、MMP-2及MMP-9蛋白表达水平(P<0.05)。结论 APC/β-catenin信号通路参与了乳腺癌细胞阿霉素耐药的发生;丹参酮IIA能够通过调控APC/β-catenin信号通路增强乳腺癌细胞对阿霉素化疗的敏感性。
Objective To assess the effect of tanshinone IIA on the chemosensitivity of doxorubicin in breast cancer cells and investigate its related mechanisms. Methods MCF-7 and MCF-7/dox cells were respectively treated with tanshinone IIA, doxorubicin, and doxorubicin combined with tanshinone IIA. MTS assay was used to detect cell proliferation; Flow cytometry was used to analyze cell apoptosis; Scratch assay was used to evaluate cell migration; Western blotting was used to detect the expressions of APC, β-catenin, E-cadherin, MMP-2, and MMP-9. Results Tanshinone IIA could significantly enhance the inhibitory effects of doxorubicin on cell proliferation and migration of MCF-7 and MCF-7/dox cells and the effect of doxorubicin on inducing cell apoptosis. Compared to MCF-7 cells, the protein expression of APC in MCF-7/dox cells was significantly decreased(P < 0.05) while the expression of β-catenin in MCF-7/dox cells was significantly increased(P < 0.05). Compared to treatment with doxorubicin alone, combined treatment of doxorubicin and tanshinone IIA could significantly up-regulate the protein expression of APC and E-cadherin(P < 0.05) and down-regulate the protein expression of β-catenin, MMP-2, and MMP-9(P < 0.05). Conclusion The APC/β-catenin pathway was involved in the development of doxorubicin resistance in breast cancer cells. Tanshinone IIA enhanced the chemosensitivity of doxorubicin in breast cancer cells through regulating APC/β-catenin pathway.
Objective To assess the effect of tanshinone IIA on the chemosensitivity of doxorubicin in breast cancer cells and investigate its related mechanisms. Methods MCF-7 and MCF-7/dox cells were respectively treated with tanshinone IIA, doxorubicin, and doxorubicin combined with tanshinone IIA. MTS assay was used to detect cell proliferation; Flow cytometry was used to analyze cell apoptosis; Scratch assay was used to evaluate cell migration; Western blotting was used to detect the expressions of APC, β-catenin, E-cadherin, MMP-2, and MMP-9. Results Tanshinone IIA could significantly enhance the inhibitory effects of doxorubicin on cell proliferation and migration of MCF-7 and MCF-7/dox cells and the effect of doxorubicin on inducing cell apoptosis. Compared to MCF-7 cells, the protein expression of APC in MCF-7/dox cells was significantly decreased(P < 0.05) while the expression of β-catenin in MCF-7/dox cells was significantly increased(P < 0.05). Compared to treatment with doxorubicin alone, combined treatment of doxorubicin and tanshinone IIA could significantly up-regulate the protein expression of APC and E-cadherin(P < 0.05) and down-regulate the protein expression of β-catenin, MMP-2, and MMP-9(P < 0.05). Conclusion The APC/β-catenin pathway was involved in the development of doxorubicin resistance in breast cancer cells. Tanshinone IIA enhanced the chemosensitivity of doxorubicin in breast cancer cells through regulating APC/β-catenin pathway.
引文
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[20]Zhan X,Li M,Zuo K,et al.upregulated mi R-155 in papillary thyroid carcinoma promotes tumor growth by targeting APC and activating Wnt/beta-catenin signaling[J].J Clin Endocrinol Metab,2013,98(8):1305-1313.
[21]Wang C,Jin H,Wang N,et al.Gas6/Axl Axis contributes to chemoresistance and metastasis in breast Cancer through Akt/GSK-3β/β-catenin signaling[J].Theranostics,2016,6(8):1205-1219.
[22]Ma J,Guo X,Zhang J,et al.PTEN gene induces cell invasion and migration via regulating AKT/GSK-3beta/beta-catenin signaling pathway in human gastric cancer[J].Dig Dis Sci,2017,60(12):3415-3425.
[23]Zhang F,Wang Z,Fan Y,et al.Elevated STAT3signaling-mediated upregulation of MMP-2/9 confers enhanced invasion ability in multidrug-resistant breast cancer cells[J].Int J Mol Sci,2015,16(10):24772-24790.
[24]李幼梅,张思仪,赵连梅,等.LIN28A、E-cadherin和vimentin在浸润性乳腺癌中的表达及其临床意义[J].癌变·畸变·突变,2017,29(4):304-308.
[25]Webb A H,Gao B T,Goldsmith Z K,et al.Inhibition of MMP-2 and MMP-9 decreasescellular migration,and angiogenesis in in vitro models of retinoblastoma[J].BMC Cancer,2017,doi:10.1186/s12885-017-3418-y.
[2]Maione F,Vincenza V,Cantone M G,et al.Molecular mechanism of tanshinone IIA and cryptotanshinone in platelet anti-aggregating effects:An integratedstudy of pharmacology and computational analysis[J].Fitoterapia,2015,doi:10.1016/j.fitote.2014.11.024.
[3]Pang H,Han B,Yu T,et al.The complex regulation of tanshinone IIA in rats with hypertension-induced left ventricular hypertrophy[J].PLo S One,2014,9(3):e92216.
[4]Chiu T L,Su C C.Tanshinone IIA increases protein expression levels of PERK,ATF6,IRE1α,CHOP,caspase-3 and caspase-12 in pancreatic cancer Bx PC-3cell-derived xenograft tumors[J].Mol Med Rep,2017,15(5):3259-3263.
[5]Li G,Shan C,Liu L,et al.Tanshinone IIA inhibits HIF-1αand VEGFA expression in breast cancer cells via m TOR/p70S6K/RPS6/4E-BP1 signaling pathway[J].PLo S One,2015,10(2):e0117440.
[6]Lee W D,Liang Y J,Chen B H.Effects of tanshinone nanoemulsion and extract on inhibition of lung cancer cells A549[J].Nanotechnology,2016,doi:10.1088/0957-4484/27/49/495101.
[7]Lu M,Wang J.Tanshinone IIA induces huamn colorectal cancer cell apoptosis:The potential roles of Aurora A-p53and survivin-mediated signaling pathways[J].Int J Oncol,2016,49(2):603-610.
[8]Huang S Y,Chang S F,Liao K F,et al.Tanshinone IIAinhibits epithelial-mesenchymal transition in bladder cancer cells via modulation of STAT3-CCL2 signaling[J].Int J Mol Sci,2017,doi:10.3390/ijms18081616.
[9]Jiang B,Zhang L,Wang Y,et al.Tanshinone IIA sodium sulfonate protects against cardiotoxicity induced by doxorubicin in vitro and in vivo[J].Food Chem Toxicol,2009,47(7):1538-1544.
[10]Hong H J,Liu J C,Chen P Y,et al.Tanshinone IIAprevents doxorubicin-induced cardiomyocyte apoptosis through Akt-dependent pathway[J].Int J Cardiol,2012,157(2):174-179.
[11]高洁,白茹,李长志,等.丹参酮IIA对阿霉素抑制肿瘤细胞生长作用的影响[J].药物评价研究,2010,33(2):91-94.
[12]Chalakur-Ramireddy N K R,Pakala S B.Combined drug therapeutic strategies for the effective treatment of triple negative breast cance[J].Biosci Rep,2018,doi:10.1042/BSR20171357.
[13]Hasan M,Leak R K,Stratford R E,et al.Drug conjugates——an emerging approach to treat breast cancer[J].Pharmacol Res Perspect,2018,6(4):e00417.
[14]Zhang X D,He C X,Cheng J,et al.Sodium tanshinone IIA sulfonate(DS-201)induces vasorelaxation of rat mesenteric arteries via inhibition of L-type Ca2+channel[J].Front Pharmacol,2018,doi:10.3389/fphar.2018.00062.
[15]冯帅,沙爽,常庆,等.丹参酮IIA对缺血性心脏病中细胞凋亡与自噬的调控机制研究进展[J].中草药,2018,49(19):4670-4677.
[16]Kanwar S S,Yu Y,Nautiyal J,et al.The Wnt/betacatenin path-way regulates growth and maintenance of colonospheres[J].Mol Cancer,2010,doi:10.1186/1476-4598-9-212.
[17]Cho S G.APC downregulated 1 inhibits breast cancer cell invasion by inhibiting the canonical WNT signaling pathway[J].Oncol Lett,2017,14(4):4845-4852.
[18]Tian J,He H,Lei G.Wnt/beta-catenin pathway in bone cancer[J].Tumour Biol,2014,35(10):9439-9445.
[19]赵轶峰,魏玉磊,王萍,等.姜黄素抑制Wnt/β-catenin信号通路诱导乳腺癌细胞凋亡的机制研究[J].现代中西医结合杂志,2016,25(29):3202-3204.
[20]Zhan X,Li M,Zuo K,et al.upregulated mi R-155 in papillary thyroid carcinoma promotes tumor growth by targeting APC and activating Wnt/beta-catenin signaling[J].J Clin Endocrinol Metab,2013,98(8):1305-1313.
[21]Wang C,Jin H,Wang N,et al.Gas6/Axl Axis contributes to chemoresistance and metastasis in breast Cancer through Akt/GSK-3β/β-catenin signaling[J].Theranostics,2016,6(8):1205-1219.
[22]Ma J,Guo X,Zhang J,et al.PTEN gene induces cell invasion and migration via regulating AKT/GSK-3beta/beta-catenin signaling pathway in human gastric cancer[J].Dig Dis Sci,2017,60(12):3415-3425.
[23]Zhang F,Wang Z,Fan Y,et al.Elevated STAT3signaling-mediated upregulation of MMP-2/9 confers enhanced invasion ability in multidrug-resistant breast cancer cells[J].Int J Mol Sci,2015,16(10):24772-24790.
[24]李幼梅,张思仪,赵连梅,等.LIN28A、E-cadherin和vimentin在浸润性乳腺癌中的表达及其临床意义[J].癌变·畸变·突变,2017,29(4):304-308.
[25]Webb A H,Gao B T,Goldsmith Z K,et al.Inhibition of MMP-2 and MMP-9 decreasescellular migration,and angiogenesis in in vitro models of retinoblastoma[J].BMC Cancer,2017,doi:10.1186/s12885-017-3418-y.