FOXC2对卵巢癌血管生成及作用机制的研究
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摘要
目的探讨叉头框C2(forkhead box C2,FOXC2)对人卵巢癌肿瘤血管生成及作用机制。方法应用FOXC2慢病毒基因转染技术,将FOXC2-LV和空载体基因分别转染到人卵巢癌A2780细胞株。细胞分为未转染组、空载体组和FOXC2过表达组。Matrigel基质胶血管形成实验和Transwell小室检测各组细胞上清作用下人脐静脉内皮细胞(HUEVCs)成管能力和迁移能力的变化。RT-PCR检测各组细胞FOXC2、DLL4/Notch1、内皮细胞粘附分子(Platelet endothelial cell adhesion molecule-1,CD31)和基质金属蛋白酶2(matrix metalloproteinase-2,MMP2)mRNA表达,Western blot检测各组细胞FOXC2、DLL4/Notch1、CD31和MMP2蛋白表达。结果 FOXC2过表达组A2780细胞上清诱导HUEVCs闭合小管数和迁移细胞数均显著高于未转染组、空载体组(P<0.05)。FOXC2过表达组DLL4/Notch1、CD31和MMP2 mRNA表达水平均显著高于未转染组和空载体组(P<0.05)。FOXC2过表达组DLL4/Notch1、CD31和MMP2 mRNA蛋白表达水平均显著高于未转染组和空载体组(P<0.05)。结论 FOXC2可能通过上调DLL4/Notch1、CD38及MMP2表达促进HUEVCs成管能力和迁移能力,影响卵巢癌发展。
Objective To investigate the effect of box C2 C2( FOXC2) on the angiogenesis and the mechanism of tumor angiogenesis in human ovarian cancer. Methods FOXC2-LV and empty vector were transfected into human ovarian cancer cell line A2780 by gene transfer technique. The cells were divided into non transfection group,empty vector group and FOXC2 over expression group. Matrigel Matrigel angiogenesis assay and Transwell assay of human umbilical vein endothelial cells cell supernatant effect( HUEVCs) changes of tube capability and migration capability. The expression levels of mRNA of FOXC2,DLL4/Notch1,detectendothelial cell adhesion molecules( Platelet endothelial cell adhesion molecule-1,CD31) and matrix metalloproteinase 2( matrix,metalloproteinase-2,MMP2) in cells of each group were detected by RT-PCR,Western expression of blot cells were detected with the expression levels of protein of FOXC2,DLL4/Notch1,CD31 and MMP2 in cells of each group were detected by western blot. Results The number and number of HUEVCs cells in A2780 cells of FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). The expression levels of mRNA of FOXC2,DLL4/Notch1,CD31 and MMP2 in FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). The protein expressions levels of FOXC2,DLL4/Notch1,CD31 and MMP2 in FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). Conclusion FOXC2 may promote expression levels of DLL4/Notch1,CD38 and HUEVCs by up regulating the MMP2 tube forming ability and migration ability,and affect the development of ovarian cancer.
Objective To investigate the effect of box C2 C2( FOXC2) on the angiogenesis and the mechanism of tumor angiogenesis in human ovarian cancer. Methods FOXC2-LV and empty vector were transfected into human ovarian cancer cell line A2780 by gene transfer technique. The cells were divided into non transfection group,empty vector group and FOXC2 over expression group. Matrigel Matrigel angiogenesis assay and Transwell assay of human umbilical vein endothelial cells cell supernatant effect( HUEVCs) changes of tube capability and migration capability. The expression levels of mRNA of FOXC2,DLL4/Notch1,detectendothelial cell adhesion molecules( Platelet endothelial cell adhesion molecule-1,CD31) and matrix metalloproteinase 2( matrix,metalloproteinase-2,MMP2) in cells of each group were detected by RT-PCR,Western expression of blot cells were detected with the expression levels of protein of FOXC2,DLL4/Notch1,CD31 and MMP2 in cells of each group were detected by western blot. Results The number and number of HUEVCs cells in A2780 cells of FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). The expression levels of mRNA of FOXC2,DLL4/Notch1,CD31 and MMP2 in FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). The protein expressions levels of FOXC2,DLL4/Notch1,CD31 and MMP2 in FOXC2 over expression group were significantly higher than those of non transfection group and empty vector group( P < 0. 05). Conclusion FOXC2 may promote expression levels of DLL4/Notch1,CD38 and HUEVCs by up regulating the MMP2 tube forming ability and migration ability,and affect the development of ovarian cancer.
引文
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[13]Wang ZQ,Bachvarova M,Morin C,et al.Role of the polypeptide N-acetylgalactosaminyltransferase 3 in ovarian cancer progression:possible implications in abnormal mucin Oglycosylation[J].Oncotarget,2014,5(2):544-560.
[2]屈洪波,吴诚义,范原铭,等.沉默FOXC2对TGF-β1诱导的MCF-7细胞上皮-间质转化的逆转作用[J].中国病理生理杂志,2013,29(5):850-856.
[3]郑春花.转录因子FOXC2在宫颈癌发生的研究[D].吉林大学,2014.
[4]冯硕.FOXC2在宫颈癌组织中的表达及其临床意义[D].吉林大学,2014.
[5]Koo HY,Shim KS,Liu T,et al.Abstract 18384:HypoxiaInduced FOXC2 Expression in Human Endothelial Cells is Dependent on the MEK and PI3K Pathways[J].Circulation,2013,6(22):18384.
[6]Sasahira T,Ueda N,Yamamoto K,et al.Prox1 and FOXC2Act as Regulators of Lymphangiogenesis and Angiogenesis in Oral Squamous Cell Carcinoma[J].Plos One,2014,9(3):92534-92534.
[7]Lin Y,Mckinnon KE,Ha SW,et al.Inorganic phosphate induces cancer cell mediated angiogenesis dependent on forkhead box protein C2(FOXC2)regulated osteopontin expression[J].Molecular Carcinogenesis,2015,54(9):926-934.
[8]Imayama N,Yamada SI,Yanamoto S,et al.FOXC2 expression is associated with tumor proliferation and invasion potential in oral tongue squamous cell carcinoma[J].Pathol Oncol Res,2015,21(3):783-791.
[9]权原.胚胎性转录因子FOXC2调控卵巢癌细胞增殖与侵袭的作用机制研究[D].吉林大学,2015.
[10]Lathia JD,Mack SC,Mulkearnshubert EE,et al.Cancer stem cells in glioblastoma.[J].Genes Dev,2015,29(12):1203-1217.
[11]Herrera AC,Victorino VJ,Campos FC,et al.Impact of tumor removal on the systemic oxidative profile of patients with breast cancer discloses lipid peroxidation at diagnosis as a putative marker of disease recurrence[J].Clin Breast Cancer.,2014,14(6):451-459.
[12]Kashatus J,Nascimento A,Myers L,et al.Erk2 Phosphorylation of Drp1 Promotes Mitochondrial Fission and MAPKDriven Tumor Growth[J].Molecular Cell,2015,57(3):537-551.
[13]Wang ZQ,Bachvarova M,Morin C,et al.Role of the polypeptide N-acetylgalactosaminyltransferase 3 in ovarian cancer progression:possible implications in abnormal mucin Oglycosylation[J].Oncotarget,2014,5(2):544-560.