表皮生长因子对胶质瘤干细胞增殖与侵袭的影响
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摘要
目的研究表皮生长因子(EGF)对胶质瘤干细胞生物学行为影响。方法胰蛋白酶消化法进行胶质瘤细胞原代培养,然后将培养基更换为含有EGF、bFGF、B27的DEME/F12培养基,培养后获得悬浮生长的肿瘤球细胞。悬浮生长的肿瘤球进行免疫荧光及分化实验。CCK-8方法检测EGF对胶质瘤干细胞增殖能力影响。Transwell方法检测EGF对胶质瘤干细胞侵袭能力影响。Western blotting实验研究EGF对胶质瘤干细胞中金属基质蛋白酶(MMP)-2表达影响。结果悬浮生长肿瘤球为胶质瘤干细胞。EGF促进胶质瘤干细胞增殖和侵袭,同时提高胶质瘤干细胞MMP-2表达。结论 EGF促进胶质瘤干细胞的增殖和侵袭与调控MMP-2信号通路相关。
Objective To study the influence of epidermal growth factor(EGF) on biological behavior of glioma stem cells. Methods The glioma cells was cultured primarily by trypsin digestion method, and then the medium was replaced by DEME/F12 medium containing EGF, BFGF and B27, and the suspended tumor spherical cells were obtained.Immunofluorescence and differentiation experiments were carried out in suspended tumor cells. The proliferation and invasion ability of glioma stem cells were detected by CCK-8 kid and Transwell respectively. The expression of metal matrix protease(MMP)-2 in glioma stem cells was studied by Western blotting. Results The suspended growth tumor cells were identified to be glioma stem cells. EGF promoted the proliferation and invasion ability of glioma stem cells and upregulated MMP-2 expression level in glioma stem cells. Conclusion The promotion of the proliferation and invasion ability of glioma stem cells by EGF was related to regulating MMP-2 signaling pathway.
Objective To study the influence of epidermal growth factor(EGF) on biological behavior of glioma stem cells. Methods The glioma cells was cultured primarily by trypsin digestion method, and then the medium was replaced by DEME/F12 medium containing EGF, BFGF and B27, and the suspended tumor spherical cells were obtained.Immunofluorescence and differentiation experiments were carried out in suspended tumor cells. The proliferation and invasion ability of glioma stem cells were detected by CCK-8 kid and Transwell respectively. The expression of metal matrix protease(MMP)-2 in glioma stem cells was studied by Western blotting. Results The suspended growth tumor cells were identified to be glioma stem cells. EGF promoted the proliferation and invasion ability of glioma stem cells and upregulated MMP-2 expression level in glioma stem cells. Conclusion The promotion of the proliferation and invasion ability of glioma stem cells by EGF was related to regulating MMP-2 signaling pathway.
引文
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[10]Ou Y,Wu Q,Wu C,et al.Migfilin promotes migration and invasion in glioma by driving EGFR and MMP-2 signalings:A positive feedback loop regulation[J].J Genet Genomics,2017,44(12):557-565.
[2]Louis DN,Perry A,Reifenberger G,et al.The 2016 World Health OrganizationClassification of Tumors of the Central Nervous System:a summary[J].Acta Neuropathol,2016,131(6):803-820.
[3]Stupp R,Hegi ME,Mason WP,et al.Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study:5-year analysis of the EORTC-NCIC trial[J].Lancet Oncol,2009,10(5):459-466.
[4]Singh SK,Hawkins C,Clarke ID,et al.Identification of human brain tumour initiating cells[J].Nature,2004,432(7015):396-401.
[5]Reya T,Morrison SJ,Clarke MF,et al.Stem cells,cancer,and cancer stem cells[J].Nature,2001,414(6859):105-111.
[6]Singh SK,Clarke ID,Terasaki M,et al.Identification of a cancer stem cell in human brain tumors[J].Cancer Res,2003,63(18):5821-5828.
[7]张东勇,王一维,张路阳,等.TGF-β2通过基质金属蛋白酶通路影响胶质瘤干细胞的侵袭能力[J].中国肿瘤生物治疗杂志,2018,25(4):357-362.
[8]邢德广,方向,王成伟,等.表皮生长因子通过上调骨桥蛋白表达促进胶质瘤细胞的增殖和侵袭[J].中华神经外科杂志,2017,33(12):1273-1277.
[9]Ramaswamy P,Aditi Devi N,Hurmath Fathima K,et al.Activation of NMDA receptor of glutamate influences MMP-2 activity and proliferation of glioma cells[J].Neurol Sci,2014,35(6):823-829.
[10]Ou Y,Wu Q,Wu C,et al.Migfilin promotes migration and invasion in glioma by driving EGFR and MMP-2 signalings:A positive feedback loop regulation[J].J Genet Genomics,2017,44(12):557-565.