VEGF-C对宫颈癌细胞增殖和凋亡信号传导通路的影响Study on the cell signal transductions of cells proliferation and apoptosis induced by VEGF-C in cervical carcinoma
杜雪;糜若然;
摘要(Abstract):
目的:研究VEGF-C对体外培养的宫颈癌HeLa细胞增殖和凋亡的影响;研究VEGF-C受体KDR、信号通路PI3K、MAPK在VEGF-C对宫颈癌增殖和凋亡调控中的作用。方法:应用重组人VEGF-C蛋白体外刺激宫颈癌HeLa细胞,MTT法检测细胞增殖,流式细胞仪检测细胞周期和凋亡,Western blot检测增殖与凋亡相关基因Bcl-2、CyclinD1蛋白表达;应用KDR-Ab、信号通路PI3K抑制剂LY294002、信号通路MAPK抑制剂PD98059预处理HeLa细胞,再进行VEGF-C刺激,观察上述指标的变化。结果:重组VEGF-C(50ng/μl)刺激HeLa细胞后增殖指数增加(2.13 vs 1),细胞周期S期比率增多[(64.26±0.20)%vs(30.91±0.09)%,P<0.05],细胞凋亡率降低(3.29±0.35 vs 7.44±0.55,P<0.05);Bcl-2、Cyclin D1表达增加(P<0.05)。KDR-Ab、LY294002预处理后与VEGF-C组相比增殖指数降低,细胞周期S期比率下降,凋亡指数升高,Bcl-2、Cyclin D1表达降低。PD98059预处理后,与VEGF-C组相比增殖指数降低、细胞周期S期比率下降、Bcl-2、Cyclin D1表达降低,但对VEGF-C诱导的凋亡无明显影响。结论:外源性VEGF-C作用于肿瘤细胞自身的KDR受体,激活细胞内信号传导通路MAPK途径和(或)PI3K途径诱导Cyclin D1表达,使肿瘤细胞S期加快,促进细胞周期的进程,进而促进He-La细胞增殖;通过PI3K途径诱导Bcl-2表达,抑制凋亡。
关键词(KeyWords): VEGF-C;宫颈癌;增殖;凋亡;信号传导
基金项目(Foundation):
作者(Authors): 杜雪;糜若然;
DOI: 10.13283/j.cnki.xdfckjz.2011.11.010
参考文献(References):
- [1]Van Trappen PO,Steele D,Lowe DG,et al.Expression of vascular endothelial growth factor(VEGF)-C and VEGF-D,and their receptor VEGFR-3,during different stages of cervical carcinogenesis[J].J Pathol,2003,201(4):544-554
- [2]Joukov V,Pajusola K,Kaipainen A,et al.A novel vascular endothelial growth factor,VEGF-C,is a ligand for the Flt4(VEGFR-3)and KDR(VEGFR-2)receptor tyrosine kina-ses[J].EMBO J,1996,15(2):290-298
- [3]Girnita L,Shenoy SK,Sehat B,et al.Beta-arrestin and Mdm2mediate IGF-1receptor-stimulated ERK activation and cell cycle progression[J].J Biol Chem,2007,282(15):11329-11338
- [4]Talha A,Yam CH,Yi S,et al.On the concentrations of cy-clins and cyclin-dependent kinases in extracts of cultured human cells[J].Biochemistry,2000,39(31):9494-9501
- [5]Lukas J,Bartkova J,Bartek J.Convergence of mitogenic signalling cascades from diverse classes of receptors at thecyclin D-cyclin-dependent kinase-pRb-controlled G1 checkpoint[J].Mol Cell Biol,1996,16(12):6917-6925
- [6]杨绍杰,孟金萍,屈祎,等.细胞凋亡信号传导通路的研究进展[J].中国比较医学杂志,2007,17(5):297-301
- [7]Dias S,Choy M,Alitalo K,et al.Vascular endothelial growth factor(VEGF)-C signaling through FLT-4(vegfr-3)mediates leukemic cell proliferation,survival,and re-sistance to chemotherapy[J].Blood,2002,99(6):2179-2184
- [8]Shibuya M,Ito N,Claesson-welsh L.Structure and function of vascular endothelial growth factor receptor-1and-2[J].Curr Top Microbiol Immunol,1999,237:59-83
- [9]Gupta K,Kshirsagar S,Li W,et al.VEGF prevents apopto-sis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling[J].Exp Cell Res,1999,247(2):495-504
- [10]Datta SR,Brunet A,Greenberg ME.Cellular survival:a play in three Akts[J].Genes Dev,1999,13(22):2905-2927
- [11]Powers MV,Workman P.Targeting of multiple signalling pathways by heat shock protein90molecular chaperone inhibitors[J].Endocr Relat Cancer,2006,13(Suppl1):S125-S135