重楼总苷Ⅰ对人视网膜色素上皮细胞系ARPE-19增殖、迁移的影响
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摘要
目的探讨重楼总苷Ⅰ对人视网膜色素上皮细胞系(ARPE-19)增生、迁移的影响。方法 MMT法检测重楼总苷Ⅰ对ARPE-19细胞增殖的影响,Transwell小室实验检测ARPE-19细胞侵袭能力的变化,划痕实验检测ARPE-19细胞迁移能力的变化,实时荧光定量PCR检测ERK1/2 mRNA的表达,Western blot法检测ERK1/2蛋白、p-ERK1/2蛋白表达水平。结果重楼总苷Ⅰ可抑制ARPE-19细胞的增生、侵袭、迁移,下调p-ERK1/2蛋白的表达,差异有统计学意义(P<0.01),但对总ERK1/2表达、ERKl mRNA和ERK2 mRNA表达无明显影响。结论重楼总苷Ⅰ能有效抑制ARPE-19细胞的增殖、侵袭、迁移,可能与重楼总苷Ⅰ抑制了ARPE-19细胞ERK1/2蛋白磷酸化有关,为中药重楼治疗增生性玻璃体视网膜疾病提供了依据。
Objective To study the effects of Polyphyllin Ⅰ on the proliferation and migration of human retinal pigment epithelial cell ARPE-19. Methods MTT assay was performed to investigate the effect of Polyphyllin I on the proliferation of ARPE-19 cells. Invasion ability of ARPE-19 cells was determined by a Matrigel invasion assay.Migration ability of ARPE-19 cells was determined by a wound healing assay. Real-time quantitative reverse transcriptase polymerase chain reaction was used to detect ERK1 /2 mRNA expression. Western blot was used to detect the expression of ERK1 /2,p-ERK1 /2. Results Polyphyllin Ⅰ inhibited proliferation of ARPE-19 cells in a dose dependent manner( P < 0. 01). Polyphyllin Ⅰ treatment reduced migration and invasion capacity of ARPE-19 cells( P < 0. 01). Polyphyllin Ⅰ treatment has no effect on the ERK1 /2 mRNA and ERK1 /2 protein expression in ARPE-19 cells,but p-ERK1 /2 protein expression was markedly decreased by treatment with Polyphyllin Ⅰ( P <0. 01). Conclusion Polyphyllin Ⅰ can effectively inhibit the ARPE-19 cells proliferation and migration,it may be related to inhibition of the phosphorylation of ERK1 /2 in ARPE-19 cells. This study provides some evidences for the efficacy of paradis in the treatment of proliferative vitreoretinopathy.
Objective To study the effects of Polyphyllin Ⅰ on the proliferation and migration of human retinal pigment epithelial cell ARPE-19. Methods MTT assay was performed to investigate the effect of Polyphyllin I on the proliferation of ARPE-19 cells. Invasion ability of ARPE-19 cells was determined by a Matrigel invasion assay.Migration ability of ARPE-19 cells was determined by a wound healing assay. Real-time quantitative reverse transcriptase polymerase chain reaction was used to detect ERK1 /2 mRNA expression. Western blot was used to detect the expression of ERK1 /2,p-ERK1 /2. Results Polyphyllin Ⅰ inhibited proliferation of ARPE-19 cells in a dose dependent manner( P < 0. 01). Polyphyllin Ⅰ treatment reduced migration and invasion capacity of ARPE-19 cells( P < 0. 01). Polyphyllin Ⅰ treatment has no effect on the ERK1 /2 mRNA and ERK1 /2 protein expression in ARPE-19 cells,but p-ERK1 /2 protein expression was markedly decreased by treatment with Polyphyllin Ⅰ( P <0. 01). Conclusion Polyphyllin Ⅰ can effectively inhibit the ARPE-19 cells proliferation and migration,it may be related to inhibition of the phosphorylation of ERK1 /2 in ARPE-19 cells. This study provides some evidences for the efficacy of paradis in the treatment of proliferative vitreoretinopathy.
引文
[1]Ryan S J.视网膜[M].天津:天津科技翻译出版公司,2011:2209-11.
[2]李彬,阚红卫,张大传,等.衰老对大鼠视网膜色素上皮细胞ROS生成和NADPH氧化酶表达的影响[J].安徽医科大学学报,2012,47(8):936-9.
[3]朱翠萍.云南红药胶囊与银杏叶片在治疗眼底出血性疾病中的应用[J].中国医药指南,2011,9(7):28-9.
[4]Kong M,Fan J,Dong A.Effects of polyphyllin I on growth inhibition of human non-small lung cancer cells and in xenograft[J].Acta Biochim Biophys Sin(Shanghai),2010,42(11):827-33.
[5]Ma D D,Lu H X,Xu L S,et al.Polyphyllin D exerts potent antitumour effects on lewis cancer cells under hypoxic conditions[J].J Int Med Res,2009,37(3):631-40.
[6]程卉,苏婧婧,候会洁,等.重楼皂苷Ⅰ抗宫颈癌He La细胞作用及机制研究[J].中药材,2013,36(11):1815-9.
[7]Khan M A,Brady C J,Kaiser R S.Clinical management of proliferative vitreoretinopathy:an update[J].Retina,2015,35(2):165-75.
[8]Dunn K C,Aotaki-Keen A E,Putkey F R,et al.ARPE-19,a human retinal pigment epithelial cell line with differentiated properties[J].Exp Eye Res,1996,62(2):155-69.
[9]陈家劲,王娟娟,张鹏,等.重楼活性单体抑制结肠癌SW620细胞增殖并诱导细胞凋亡[J].暨南大学学报(自然科学与医学版),2015,36(2):124-30.
[10]Marshall C J.Specificity of receptor tyrosine kinase signaling:transient versus sustained extracellular signal-regulated kinase activation[J].Cell,1995,80(2):179-85.
[11]Yokawa E,Takai S,Tanaka M,et al.Overexpression of ERK,an EPH family receptor protein tyrosine kinase,in various human tumors[J].Cancer Res,1994,54(14):3645-50.
[12]Lim S C,Duong H Q,Parajuli K R,et al.Pro-apoptotic role of the MEK/ERK pathway in urso-deoxycholic acid-induced apoptosis in SNU601 gastric cancer cells[J].Oncol Rep,2012,28(4):1429-34.
[13]Ling M,Li Y,Xu Y,et al.Regulation of miRNA-21 by reactive oxygen species-activated ERK/NF-κB in arsenite-induced cell transformation[J].Free Radic Biol Med,2012,52(9):1508-18.
[14]Huang T H,Wu F,Loeb G B,et al.Up-regulation of miR-21 by HER2/neu signaling promotes cell invasion[J].J Biol Chem,2009,284(27):18515-24.
[2]李彬,阚红卫,张大传,等.衰老对大鼠视网膜色素上皮细胞ROS生成和NADPH氧化酶表达的影响[J].安徽医科大学学报,2012,47(8):936-9.
[3]朱翠萍.云南红药胶囊与银杏叶片在治疗眼底出血性疾病中的应用[J].中国医药指南,2011,9(7):28-9.
[4]Kong M,Fan J,Dong A.Effects of polyphyllin I on growth inhibition of human non-small lung cancer cells and in xenograft[J].Acta Biochim Biophys Sin(Shanghai),2010,42(11):827-33.
[5]Ma D D,Lu H X,Xu L S,et al.Polyphyllin D exerts potent antitumour effects on lewis cancer cells under hypoxic conditions[J].J Int Med Res,2009,37(3):631-40.
[6]程卉,苏婧婧,候会洁,等.重楼皂苷Ⅰ抗宫颈癌He La细胞作用及机制研究[J].中药材,2013,36(11):1815-9.
[7]Khan M A,Brady C J,Kaiser R S.Clinical management of proliferative vitreoretinopathy:an update[J].Retina,2015,35(2):165-75.
[8]Dunn K C,Aotaki-Keen A E,Putkey F R,et al.ARPE-19,a human retinal pigment epithelial cell line with differentiated properties[J].Exp Eye Res,1996,62(2):155-69.
[9]陈家劲,王娟娟,张鹏,等.重楼活性单体抑制结肠癌SW620细胞增殖并诱导细胞凋亡[J].暨南大学学报(自然科学与医学版),2015,36(2):124-30.
[10]Marshall C J.Specificity of receptor tyrosine kinase signaling:transient versus sustained extracellular signal-regulated kinase activation[J].Cell,1995,80(2):179-85.
[11]Yokawa E,Takai S,Tanaka M,et al.Overexpression of ERK,an EPH family receptor protein tyrosine kinase,in various human tumors[J].Cancer Res,1994,54(14):3645-50.
[12]Lim S C,Duong H Q,Parajuli K R,et al.Pro-apoptotic role of the MEK/ERK pathway in urso-deoxycholic acid-induced apoptosis in SNU601 gastric cancer cells[J].Oncol Rep,2012,28(4):1429-34.
[13]Ling M,Li Y,Xu Y,et al.Regulation of miRNA-21 by reactive oxygen species-activated ERK/NF-κB in arsenite-induced cell transformation[J].Free Radic Biol Med,2012,52(9):1508-18.
[14]Huang T H,Wu F,Loeb G B,et al.Up-regulation of miR-21 by HER2/neu signaling promotes cell invasion[J].J Biol Chem,2009,284(27):18515-24.