迷迭香酸抑制人舌癌Tca8113细胞转移作用及初步机制研究
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摘要
目的探讨迷迭香酸对舌癌Tca8113细胞转移的抑制作用及初步机制。方法用磺酰罗丹明B(SRB)比色法检测迷迭香酸对Tca8113细胞增殖活性的影响;Transwell小室迁移试验测定舌癌Tca8113细胞的迁移活性;明胶酶谱法检测Tca8113细胞中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)的表达活性;蛋白质印迹法检测蛋白IKK、p-IKK、Akt和p-Akt在Tca8113细胞中表达的变化。结果迷迭香酸能明显抑制Tca8113细胞的增殖活性,且具有时间和浓度依赖关系;能浓度依赖性降低Tca8113细胞的迁移活性以及细胞中MMP-2和MMP-9的表达(P<0.05)。胞浆中蛋白IKK的总量和p-IKK的量均随着迷迭香酸浓度的增加而降低;Akt的总量并不随迷迭香酸的浓度而变化,但是p-Akt的量明显呈浓度依赖性降低(P<0.05)。结论迷迭香酸对舌癌Tca8113细胞的转移具有一定的抑制作用,其机制可能与PI3K/Akt信号通路和核因子-κB信号通路有关。
Objective To investigate the inhibitory effects of RA(rosmarinic acid)on metastasis of human tongue carcinoma Tca8113 cells and its preliminary mechanisms. Methods The inhibitory effects of RA onthe proliferation of Tca8113 cells were observed using sulforhodamine B(SRB) assay. Transwell chamber assay was done to study the inhibitory effects of RA on the migration of Tca8113 cells. The expressions of MMP-2 and MMP-9 in Tca8113 cells were assessed through Gelatin Zymography assay. The protein expressions of IKK, p-IKK, Akt and p-Akt were committed using Western blotting analysis. Results RA significantly inhibited the proliferation of Tca8113 cells in a concentration- and time- dependent manner. Meanwhile,RA also reduced the migration activities and MMP-2 and MMP-9 expressions of Tca8113 cells in a concentration-dependent manner. Moreover, the IKK and p-IKK expressions in cytoplasm for Tca8113 cells were reduced in a concentration-dependent manner, but the expressions of Akt did not changed by RA, and the level of p-Akt was reduced in a concentration-dependent manner. Conclusion RA can inhibit the proliferation, migration and expressions of MMP- 2 and MMP- 9 in Tca8113 cells and the mechanisms maybe attribute to PI3K/Akt and NF-κB signaling pathway.
Objective To investigate the inhibitory effects of RA(rosmarinic acid)on metastasis of human tongue carcinoma Tca8113 cells and its preliminary mechanisms. Methods The inhibitory effects of RA onthe proliferation of Tca8113 cells were observed using sulforhodamine B(SRB) assay. Transwell chamber assay was done to study the inhibitory effects of RA on the migration of Tca8113 cells. The expressions of MMP-2 and MMP-9 in Tca8113 cells were assessed through Gelatin Zymography assay. The protein expressions of IKK, p-IKK, Akt and p-Akt were committed using Western blotting analysis. Results RA significantly inhibited the proliferation of Tca8113 cells in a concentration- and time- dependent manner. Meanwhile,RA also reduced the migration activities and MMP-2 and MMP-9 expressions of Tca8113 cells in a concentration-dependent manner. Moreover, the IKK and p-IKK expressions in cytoplasm for Tca8113 cells were reduced in a concentration-dependent manner, but the expressions of Akt did not changed by RA, and the level of p-Akt was reduced in a concentration-dependent manner. Conclusion RA can inhibit the proliferation, migration and expressions of MMP- 2 and MMP- 9 in Tca8113 cells and the mechanisms maybe attribute to PI3K/Akt and NF-κB signaling pathway.
引文
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[15]Wu H T,Sie S S,Kuan T C,et al.Identifying the regulative role of NF-kappa B binding sites within promoter region of human matrix metalloproteinase 9(MMP-9)by TNF-alpha induction[J].Appl Biochem Biotechnol,2013,169(2):438-449.
[2]Xu Y,Xu G,Liu L,et al.Anti-invasion effect of rosmarinic acid via the extracellular singnal-regulated kinase and exidation-reduction pathway in Ls174-T cells[J].J Cell Biochem,2010,111(2):370-379.
[3]Xu Y,Jiang Z,Ji G,et al.Inhibition of bone metastasis from breast carcinoma by rosmarinic acid[J].Planta Med,2010,76(10):956-962.
[4]Jeong H J,Choi Y,Kim M H,et al.Rosmarinic acid,active component of Dansam-Eum attenuates ototoxicity of cochlear hair cells through blockage of caspase-1 activity[J].PLo S One,2011,6(4):e18815.
[5]王志强,张秀英,李文广,等.异甘草素抗肿瘤活性及初步机制研究[J].中国药理学通报,2015,31(8):1159-1165.
[6]Zhang X Y,Zhu P,Zhang X Y,et al.Natural antioxidantisoliquiritigenin ameliorates contractile dysfunction of hypoxic cardiomyocytes via AMPK signaling pathway[J].Mediators Inflamm,2013,2013:390-890.
[7]Spallarossa P,Garibaldi S,Altieri P,et al.Carvedilol prevents doxorubicin-induced free radical release and apoptosis in cardiomyocytes in vitro[J].J Mol Cell Cardiol,2004,37(4):837-846.
[8]Yesil-Celiktas O,Sevimli C,Bedir E,et al.Inhibitory effects of rosemary extracts,carnosic acid and rosmarinic acid on the growth of various human cancer cell lines[J].Plant Foods Hun Nutr,2010,65(2):158-163.
[9]Tavafi M,Ahmadvand H.Effect of rosmarinic acid on inhibition of gentamicin induced nephrotoxicity in rats[J].Tissue Cell,2011,43(6):392-397.
[10]Ishigami S,Natsugoe S,Nakashima H,et al.Biological aggressiveness of alpha-fetoprotein(AFP)-positive gastric cancer[J].Hepatogasroenterology,2006,53(69):338-341.
[11]Katori H,Baba Y,Imagawa Y,et al.Reduction of in vivo tumor growth by MMI-166,a selective matrix metalloproteinase inhibitor,through inhibition of tumor angiogenesis in squamous cell carcinoma cell lines of head and neck[J].Cancer Letters,2002,178(2):151-159.
[12]Zhou X,Tna M I,Stone Hawthorne V,et al.Activation of the Akt/mammalian target of rapamycin/4E-BP1 pathway by Erb B2 overexpression predicts tumor progression in breast cancers[J].Clin Cancer Res,2004,10(20):6779-6788.
[13]Dan H C,Cooper M J,Cogswell P C,et al.Akt-dependent regulation of NF-kappa B is controlled by m TOR and Raptor in association with IKK[J].Genes Dev,2008,22(11):1490-1500.
[14]Molinero L L,Alegre M L.Role of T cell-nuclear factorκB in transplantation[J].Transplant Rev(Orlando),2012,26(3):189-200.
[15]Wu H T,Sie S S,Kuan T C,et al.Identifying the regulative role of NF-kappa B binding sites within promoter region of human matrix metalloproteinase 9(MMP-9)by TNF-alpha induction[J].Appl Biochem Biotechnol,2013,169(2):438-449.