摘要
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。
Objective：To investigate cancer cells proliferation and apoptosis regulated by VEGF-C on HeLa cell line.To confirm whether KDR,a receptor of VEGF-C,and cell transduction pathways PI3K or MAPK were involved in the proliferation and apoptosis induced by VEGF-C in vivo.Methods：Stimulated by recombinant VEGF-C in vivo,HeLa cells were used to analyze the index of cell proliferation by MTT assay.Early apoptosis and cell cycle were analyzed by FCM and proteins expression of Bcl-2 and Cyclin D1 by Western Blotting.HeLa cells were treated by VEGF-C,KDR antibody,MAPK specific inhibitor PD98059,and PI3K specific inhibitor LY294002,and the effects were the investigated through the above-mentioned applications.Results：Treatment of VEGF-C（50ng/μl）was found to increase cells proliferation（2.13 vs 1.00）,cells number in S phase（64.26±0.20 vs 30.91±0.09）,and apoptosis index significantly reduced after the stimulation of VEGF-C（5.5±0.25 vs 12.4±0.30）.The protein expression level of Bcl-2 and Cyclin D1 in HeLa were increased after stimulated by VEGF-C.The effects induced by VEGF-C including cells proliferation,apoptosis inhibition,the Bcl-2 and Cyclin D1 expression were blocked in certain degree by Pre-simulated with KDR-Ab and LY294002 as well as PD98059.But apoptosis inhibition effect induced by VEGF-C was not influenced by PD98059.Conclusions：In response to exogenous VEGF-C,the cells proliferation is increased as mediated by KDR,which involved the activation of PI3K or/and MAPK pathways through increasing the expression of Cyclin D1 to promote cells cycle phase S.Cell apoptosis is inhibited through cell signal transduction PI3K by increasing the expression of Bcl-2.
出处
《现代妇产科进展》
CSCD
北大核心
2011年第11期877-880,885,共5页
Progress in Obstetrics and Gynecology
关键词
VEGF-C
宫颈癌
增殖
凋亡
信号传导
VEGF-C
Cervical cancer
Proliferation
Apoptosis
Cells signal transduction
