人参皂苷Rh2对神经母细胞瘤SH-SY5Y的增殖、凋亡、迁移及侵袭的影响
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摘要
目的:针对神经母细胞瘤(neuroblastoma,NB)早期易转移的特性,研究人参皂苷Rh2(ginsenoside Rh2)对神经母细胞瘤的增殖、凋亡、迁移及侵袭的影响。方法:用不同(5、10、20、40、80μg/ml)浓度人参皂苷Rh2来干预神经母细胞瘤细胞系SH-SY5Y。Cell counting kit-8(CCK-8)法、细胞划痕试验、Transwell小室模型检测细胞增殖、迁移及侵袭能力;蛋白质印迹法(Western-blot,WB)试验检测不同浓度下神经母细胞瘤细胞内基质金属蛋白酶2(matrix metalloproteinase2,MMP2)、Bax、Bcl-2、β-catenin蛋白的表达。结果:人参皂苷Rh2可以时间-浓度依赖性地抑制神经母细胞瘤增殖、迁移及侵袭,降低迁移相关蛋白MMP2的表达,上调了促凋亡蛋白Bax的表达水平同时降低抗凋亡蛋白Bcl-2的表达。人参皂苷Rh2处理的神经母细胞瘤内β-catenin水平降低。结论:人参皂苷Rh2可以在体外抑制神经母细胞瘤的增殖,下调MMP2蛋白抑制肿瘤细胞迁移及侵袭,通过wnt通路抑制细胞生长。
Objective: To investigate the characteristics of early metastasis of neuroblastoma(NB),the effects of ginsenoside Rh2 on proliferation,apoptosis,migration and invasion of neuroblastoma were studied.Methods: Ginsenoside Rh2 was used to interfere with neuroblastoma cell line SH-SY5Y(5,10,20,40,80 μg/ml).Cell counting kit -8(CCK-8) assay was used to detect cell proliferation,cell scratch test was used to detect cell migration ability,transwell cell model was used to detect cell invasiveness,and Western-blot test was used to detect the expression of matrix metalloproteinase 2(MMP2) Bax,Bcl-2,and β-catenin protein in neuroblastoma cells under different concentrations.Results: Ginsenoside Rh2 can inhibit the proliferation,migration and invasion of neuroblastoma in time and dose dependent manner to reduce the expression of migration related protein MMP2.The level of β-catenin in neuroblastoma cells treated with ginsenoside Rh2 decreased.Conclusion: Ginsenoside Rh2 can inhibit the proliferation of neuroblastoma cells in vitro,and down regulation of MMP2 protein to inhibit the migration and invasion of tumor cells,and inhibit cell growth through wnt pathway.
Objective: To investigate the characteristics of early metastasis of neuroblastoma(NB),the effects of ginsenoside Rh2 on proliferation,apoptosis,migration and invasion of neuroblastoma were studied.Methods: Ginsenoside Rh2 was used to interfere with neuroblastoma cell line SH-SY5Y(5,10,20,40,80 μg/ml).Cell counting kit -8(CCK-8) assay was used to detect cell proliferation,cell scratch test was used to detect cell migration ability,transwell cell model was used to detect cell invasiveness,and Western-blot test was used to detect the expression of matrix metalloproteinase 2(MMP2) Bax,Bcl-2,and β-catenin protein in neuroblastoma cells under different concentrations.Results: Ginsenoside Rh2 can inhibit the proliferation,migration and invasion of neuroblastoma in time and dose dependent manner to reduce the expression of migration related protein MMP2.The level of β-catenin in neuroblastoma cells treated with ginsenoside Rh2 decreased.Conclusion: Ginsenoside Rh2 can inhibit the proliferation of neuroblastoma cells in vitro,and down regulation of MMP2 protein to inhibit the migration and invasion of tumor cells,and inhibit cell growth through wnt pathway.
引文
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[16]Matthay KK,Reynolds CP,Seeger RC,et al.Long-term results for children with high-risk neuroblastoma treated on a randomized trial of myeloablative therapy followed by 13-cis-retinoic acid:a children's oncology group study[J].Journal of Clinical Oncology,2009,27(7):1007-1013.
[17]Cheung NK,Kushner BH,Laquaglia M,et al.N7:a novel multimodality therapy of high risk neuroblastoma(NB)in children diagnosed over 1 year of age[J].Medical and Pediatric Oncology,2001,36(1):227-230.
[18]Yang Z,Zhao T,Liu H,et al.Ginsenoside Rh2 inhibits hepatocellular carcinoma through beta-catenin and autophagy[J].Scientific Reports,2016,6:19383.
[19]Qian J,Li J,Jia JG,et al.Ginsenoside-Rh2 inhibits proliferation and induces apoptosis of human gastric cancer SGC-7901 side population cells[J].Asian Pacific Journal of Cancer Prevention:APJCP,2016,17(4):1817-1821.
[20]Choi S,Kim TW,Singh SV.Ginsenoside Rh2-mediated G1phase cell cycle arrest in human breast cancer cells is caused by p15 Ink4B and p27 Kip1-dependent inhibition of cyclin-dependent kinases[J].Pharmaceutical Research,2009,26(10):2280-2288.
[21]Somiari SB,Somiari RI,Heckman CM,et al.Circulating MMP2and MMP9 in breast cancer-potential role in classification of patients into low risk,high risk,benign disease and breast cancer categories[J].International Journal of Cancer,2006,119(6):1403-1411.
[22]Tutton MG,George ML,Eccles SA,et al.Use of plasma MMP-2and MMP-9 levels as a surrogate for tumour expression in colorectal cancer patients[J].International Journal of Cancer,2003,107(4):541-550.
[2]Cheung NK,Dyer MA.Neuroblastoma:developmental biology,cancer genomics and immunotherapy[J].Nature Reviews Cancer,2013,13(6):397-411.
[3]Vo KT,Matthay KK,Neuhaus J,et al.Clinical,biologic,and prognostic differences on the basis of primary tumor site in neuroblastoma:A report from the international neuroblastoma risk group project[J].Journal of Clinical Oncology,2014,32(28):3169-3176.
[4]Schulte M,Koster J,Rahmann S,et al.Cancer evolution,mutations,and clonal selection in relapse neuroblastoma[J].Cancer,2018,372(2):263-268.
[5]Guan N,Huo X,Zhang Z,et al.Ginsenoside Rh2 inhibits metastasis of glioblastoma multiforme through Akt-regulated MMP13[J].Tumour Biology,2015,36(9):6789-6795.
[6]Zammit V,Baron B,Ayers D.Mi RNA influences in neuroblast modulation:An introspective analysis[J].Genes,2018,9(1):26.
[7]Liao YX,Zhang ZP,Zhao J,et al.Effects of fibronectin 1 on cell proliferation,senescence and apoptosis of human glioma cells through the PI3K/AKT signaling Pathway[J].Cellular Physiology and Biochemistry,2018,48(3):1382-1396.
[8]Wang W,Li P,Xu J,et al.Resveratrol attenuates high glucose-induced nucleus pulposus cell apoptosis and senescence through activating the ROS-mediated PI3K/Akt pathway[J].Bioscience Reports,2018,38(2):BSR20171454.
[9]Zhuang RJ,Ma J,Shi X,et al.Cold-inducible protein RBM3 protects UV irradiation-induced apoptosis in neuroblastoma cells by affecting p38 and JNK pathways and Bcl2 family proteins[J].2017,63(2):142-151.
[10]Kim YS,Jin SH.Ginsenoside Rh2 induces apoptosis via activation of caspase-1 and-3 and up-regulation of Bax in human neuroblastoma[J].Archives of Pharmacal Research,2004,27(8):834-839.
[11]Fei XF,Wang BX,Tashiro S,et al.Apoptotic effects of ginsenoside Rh2 on human malignant melanoma A375-S2 cells[J].Acta Pharmacologica Sinica,2002,23(4):315-322.
[12]Kim YS,Jin SH,Lee YH,et al.Ginsenoside Rh2 induces apoptosis independently of Bcl-2,Bcl-x L,or Bax in C6Bu-1 cells[J].Archives of Pharmacal Research,1999,22(5):448-453.
[13]Tolbert VP,Coggins GE,Maris JM.Genetic susceptibility to neuroblastoma[J].Current Opinion in Genetics&Development,2017,42:81-90.
[14]Maris JM,Hogarty MD,Bagatell R,et al.Neuroblastoma[J].Lancet(London,England),2007,369(9579):2106-2120.
[15]Xicoy H,Wieringa B,Martens GJ.The SH-SY5Y cell line in Parkinson's disease research:a systematic review[J].Molecular Neurodegeneration,2017,12(1):10.
[16]Matthay KK,Reynolds CP,Seeger RC,et al.Long-term results for children with high-risk neuroblastoma treated on a randomized trial of myeloablative therapy followed by 13-cis-retinoic acid:a children's oncology group study[J].Journal of Clinical Oncology,2009,27(7):1007-1013.
[17]Cheung NK,Kushner BH,Laquaglia M,et al.N7:a novel multimodality therapy of high risk neuroblastoma(NB)in children diagnosed over 1 year of age[J].Medical and Pediatric Oncology,2001,36(1):227-230.
[18]Yang Z,Zhao T,Liu H,et al.Ginsenoside Rh2 inhibits hepatocellular carcinoma through beta-catenin and autophagy[J].Scientific Reports,2016,6:19383.
[19]Qian J,Li J,Jia JG,et al.Ginsenoside-Rh2 inhibits proliferation and induces apoptosis of human gastric cancer SGC-7901 side population cells[J].Asian Pacific Journal of Cancer Prevention:APJCP,2016,17(4):1817-1821.
[20]Choi S,Kim TW,Singh SV.Ginsenoside Rh2-mediated G1phase cell cycle arrest in human breast cancer cells is caused by p15 Ink4B and p27 Kip1-dependent inhibition of cyclin-dependent kinases[J].Pharmaceutical Research,2009,26(10):2280-2288.
[21]Somiari SB,Somiari RI,Heckman CM,et al.Circulating MMP2and MMP9 in breast cancer-potential role in classification of patients into low risk,high risk,benign disease and breast cancer categories[J].International Journal of Cancer,2006,119(6):1403-1411.
[22]Tutton MG,George ML,Eccles SA,et al.Use of plasma MMP-2and MMP-9 levels as a surrogate for tumour expression in colorectal cancer patients[J].International Journal of Cancer,2003,107(4):541-550.