L1CAM及α5-整合素表达对胰腺癌细胞分泌促血管生成因子的影响  

Effect of the expression of L1CAM and α5-integrin to the secrete of pro-angiogenic factors in pancreatic cancer

作　　者：王奕智[1] 葛春林[1] 

机构地区：[1]中国医科大学附属第一医院胰胆外科,辽宁沈阳110001

出　　处：《现代肿瘤医学》2018年第7期998-1004,共7页Journal of Modern Oncology

基　　金：2015年沈阳市科学技术计划项目(编号:F15-199-1-48);2014年辽宁省教育厅(编号:L2014294)

摘　　要：目的:探究体外培养胰腺癌细胞系中L1CAM与α5-整合素产生促血管生成因子对内皮细胞血管生成方面的影响。方法:将重组质粒pc DNA3.1-L1CAM和α5-整合素小干扰RNA分别转入Patu8988和Capan-2胰腺癌细胞系上调和下调靶基因的表达。Western blot及实时定量PCR检测Patu8988及Capan-2胰腺癌细胞系中促血管生成因子[信号转导及转录激活因子3(signal transducers and activators of transcription3,STAT3)、磷酸化信号转导及转录因子3(phosphorylated signal transducers and activators of transcription 3,pSTAT3)、血管内皮生长因子-A(vascular endothelial growth factor-A,VEGF-A)、血管生成素2(angiopoietin-2,Ang-2)]表达的水平差异;血管生成实验检测改变L1CAM及α5-整合素表达水平后Patu8988和Capan-2胰腺癌细胞系上清液对体外血管生成的影响。结果:上调Patu8988及Capan-2胰腺癌细胞系中L1CAM的表达水平可促进pSTAT3、VEGF-A及Ang-2的mRNA及蛋白表达水平(P<0.01),并促进人脐静脉内皮细胞系(human umbilical vein endothelium cell,HUVEC)的完整血管形成(P<0.01)。而抑制Patu8988及Capan-2胰腺癌细胞系中的α5-整合素表达水平显著减少pSTAT3、VEGF-A及Ang-2的mRNA及蛋白表达水平(P<0.01),并明显削减了HUVEC的成管能力(P<0.01)。结论:体外培养胰腺癌细胞系中,L1CAM表达水平增加显著促进促血管生成因子表达和血管生成能力;而抑制α5-整合素的表达水平则可显著抑制促血管生成因子表达和成管能力。Objective：To determine the promoted effect of the pro-angiogenic factors produced by L1 cell adhesion molecule （L1CAM） and α5-integrin on tube formation of endothelium in pancreatic cancer in vitro.Methods：pcDNA3.1-L1CAM and α5-integrin siRNA were transfected into Patu8988 and Capan-2 cell lines respectively to upregulate and downregulate the level of target proteins.Western blot and qRT-PCR were used to detect the difference levels between experimental group and control group about pro-angiogenic factors in Patu8988 and Capan-2 cell lines,such as signal transducers and activators of transcription 3 （STAT3）,phosphorylated signal transducers and activators of transcription 3 （pSTAT3）,vascular endothelial growth factor-A （VEGF-A） and angiopoietin-2 （Ang-2）.Tube formation assay was used to determine the effect of supernatant of Patu8988 and Capan-2 cell lines to capacity of tube formation after transfecting pcDNA3.1-L1CAM and α5-integrin siRNA.Results：High level of L1CAM promoted the most network formation of human umbilical vein endothelium cell （HUVEC）（P〈0.01） and increased the mRNA and protein level of pSTAT3,VEGF-A and Ang-2 （P〈0.01）.Silencing the expression of α5-integrin inhibited the ability of tube formation of HUVEC （P〈0.01） and decreased the mRNA and protein level of pSTAT3,VEGF-A and Ang-2 （P〈0.01）.Conclusion：In pancreatic cancer cell lines in vitro,increased level of L1CAM induced the expression of the pro-angiogenic factors and tube formation,and silencing the expression of α5-integrin reduced the expression of the pro-angiogenic factors and tube formation.

关 键 词：胰腺癌 L1CAM 血管生成 α5-整合素 

分 类 号：R735.9[医药卫生—肿瘤]