羊栖菜多糖对人肺癌SPC-A-1细胞及裸鼠移植瘤Ecadherin、MMP2和MMP9表达的影响
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摘要
目的:观察羊栖菜多糖(Sargassum fusiforme polysaccharides,SFPS)对体外培养的人肺癌细胞株SPC-A-1及裸鼠移植瘤组织中上皮钙粘蛋白(Epithelial cadherin,Ecadherin)、基质金属蛋白酶(Matrix metalloproteinase,MMP)2、9表达的影响。方法:应用实时细胞分析技术检测SPC-A-1细胞增殖活性,采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞培养液中Ecadherin、MMP2和MMP9含量;建立裸鼠移植瘤模型,随机分组,采用免疫组化法检测肿瘤组织中Ecadherin、MMP2和MMP9表达。结果:与对照组相比,30、100和300mg/L浓度SFPS作用24h即可抑制SPC-A-1细胞增殖,作用36小时抑制效果最为明显(P<0.05,P<0.01,P<0.001);SFPS作用36h后,100mg/L、300mg/L浓度组的细胞培养液中Ecadherin含量明显增加(P<0.05,P<0.001),而MMP2和MMP9含量在各浓度组(30、100和300mg/L)均降低(P<0.05,P<0.01,P<0.001);20、40mg/kg剂量SFPS处理组的裸鼠移植瘤组织中Ecadherin表达显著增加(P<0.001),而MMP2和MMP9表达明显低于对照组(P<0.001)。结论:SFPS可以抑制SPC-A-1肺癌细胞生长,此作用可能与下调MMP2、MMP9表达水平、上调Ecadherin表达水平有关,提示SFPS具有潜在的抗肿瘤转移作用。
Objective: To observe the effect of Sargassum Fusiforme Polysaccharides(SFPS) on the expression of epithelial cadherin(Ecadherin),matrix metalloproteinase(MMP) 2 and 9 in human lung cancer SPC-A-1 cells and xenografts of nude mice. Methods: Real-time cell analysis was utilized to evaluate the proliferation of SPC-A-1 cells. Ecadherin,MMP2 and MMP9 level in the supernatant was measured by enzyme linked immunosorbent assay(ELISA). Models of xenografts in nude mice were established,and immunohistochemical method was applied to detect the expression of Ecadherin,MMP2 and MMP9 in xenografts. Results: After treatment of 30,100 mg/L and 300 mg/L SFPS on SPC-A-1 cells for 24 h,the proliferation of cells was inhibited,and the effect was the most significant after treated by SFPS for 36 h compared with the controlled group(P<0.05,P<0.01,P<0.001). Ecadherin in the supernatant of SPC-A-1 cells was increased after treated by 100,300 mg/L SFPS for 36 h(P<0.05,P<0.001),while MMP2 and MMP9 were obviously decreased in 30,100 and 300 mg/L SFPS groups(P<0.05,P<0.01,P<0.001). The expression of Ecadherin was markedly enhanced in xenografts of nude mice treated by 20,40 mg/kg SFPS(P<0.001),while MMP2 and MMP9 were obviously reduced(P<0.001). Conclusions: SFPS can inhibit the proliferation of SPC-A-1 cells through down-regulating the expression of MMP2,MMP9 and up-regulating the expression of Ecadherin,which suggest that SFPS might restrain the metastasis of cancer cells.
Objective: To observe the effect of Sargassum Fusiforme Polysaccharides(SFPS) on the expression of epithelial cadherin(Ecadherin),matrix metalloproteinase(MMP) 2 and 9 in human lung cancer SPC-A-1 cells and xenografts of nude mice. Methods: Real-time cell analysis was utilized to evaluate the proliferation of SPC-A-1 cells. Ecadherin,MMP2 and MMP9 level in the supernatant was measured by enzyme linked immunosorbent assay(ELISA). Models of xenografts in nude mice were established,and immunohistochemical method was applied to detect the expression of Ecadherin,MMP2 and MMP9 in xenografts. Results: After treatment of 30,100 mg/L and 300 mg/L SFPS on SPC-A-1 cells for 24 h,the proliferation of cells was inhibited,and the effect was the most significant after treated by SFPS for 36 h compared with the controlled group(P<0.05,P<0.01,P<0.001). Ecadherin in the supernatant of SPC-A-1 cells was increased after treated by 100,300 mg/L SFPS for 36 h(P<0.05,P<0.001),while MMP2 and MMP9 were obviously decreased in 30,100 and 300 mg/L SFPS groups(P<0.05,P<0.01,P<0.001). The expression of Ecadherin was markedly enhanced in xenografts of nude mice treated by 20,40 mg/kg SFPS(P<0.001),while MMP2 and MMP9 were obviously reduced(P<0.001). Conclusions: SFPS can inhibit the proliferation of SPC-A-1 cells through down-regulating the expression of MMP2,MMP9 and up-regulating the expression of Ecadherin,which suggest that SFPS might restrain the metastasis of cancer cells.
引文
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[2] Ji YB,Ji CF,Yue L.Human gastric cancer cell line SGC-7901apoptosis induced by SFPS-B2via a mitochondrial-mediated pathway[J].Biomed Mater Eng,2014,24(1):1141～1147
[3] 严磷磷,梁倩,李继承,等.羊栖菜多糖通过激活Caspase途径诱导Lovo细胞凋亡[J].细胞生物学杂志,2006,28(2):193～200
[4] Chen X,Nie W,Yu G,et al.Antitumor and immunomodulatory activity of polysaccharides from Sargassum fusiforme[J].Food Chem Toxicol,2012,50(3～4):695～700
[5] Chen H,hang L,Long X,et al.Sargassum fusiforme polysaccharides inhibit VEGF-A-related angiogenesis and proliferation of lung cancer in vitro and in vivo[J].Biomed Pharmacother,2017,85:22～27
[6] Wu X,Jiang W,Lu J,Yu Y,Wu B.Analysis of the monosaccharide composition of water-soluble polysaccharides from Sargassum fusiforme by high performance liquid chromatography/electrospray ionisation mass spectrometry[J].Food chemistry,2014,145:976～983
[7] 王百龙,孙稚颖,周凤琴.羊栖菜成分以及药理研究进展[J].山东中医杂志,2015,34(9):722～725
[8] 梁倩,李继承,张华芳.羊栖菜多糖诱导HL-60细胞凋亡的研究[J].实验生物学报,2004,37(2):125～128
[9] 王建光,杨新宇.羊栖菜多糖诱导MCF-7细胞凋亡机制的研究[J].中国老年学杂志,2005,25(5):567～569
[10] 况炜,陈慧玲.羊栖菜多糖对人肺癌细胞及肿瘤血管内皮细胞模型增殖活性的影响[J].现代实用医学,2011,23(3):256～257,267
[11] Gialeli C,Theocharis AD,Karamanos NK.Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting[J].FEBS J,2011,278(1):16～27
[12] Solov’eva NI,Timoshenko OS,Kugaevskaia EV,et al.Key enzymes of degradation and angiogenesis as a factors of tumor progression in squamous cell carcinoma of the cervix[J].Bioorg Khim,2014,40(6):743～751
[13] 史芳涛,王庆锋.凋亡抑制蛋白基因Livin及肿瘤转移相关基因在肺癌组织中的表达及其与肺癌临床特征的关系分析[J].癌症进展,2017,15(7):794～797,805
[14] Ii M,Yamamoto H,Adachi Y,et al.Role of matrix metalloproteinase-7(matrilysin) in human cancer invasion,apoptosis,growth,and angiogenesis[J].Exp Biol Med (Maywood).2006,231(1):20～27
[15] 郝立宏,赵瑾瑶,宫琳琳,等.上皮钙黏蛋白在肺癌组织中的表达及其基因点突变[J].解剖学杂志,2012,35(2):171～173,185