轴突诱导因子4D对人胰腺癌细胞增殖、迁移及血管生成的影响  

Effect of semaphorin 4D on the proliferation,migration and angiogenic of human pancreatic carcinoma cells

作　　者：贾如江[1] 侯丽艳[1] 尹清臣[1] 李丽芳 杨庚武[1] 

机构地区：[1]邯郸市中心医院普外一科,河北邯郸056102 [2]邯郸市丛台区人民医院外科,河北邯郸056102

出　　处：《新乡医学院学报》2017年第12期1063-1067,共5页Journal of Xinxiang Medical University

基　　金：河北省卫计委计划项目(编号:20150457)

摘　　要：目的探讨siRNA沉默轴突诱导因子4D(Sema4D)对人胰腺癌细胞增殖、迁移及血管生成的影响。方法设计合成Sema4D-siRNA,转染至人胰腺癌细胞系中,瞬时转染48 h后利用反转录-聚合酶链反应检测转染前后Sema4D mRNA表达的变化;瞬时转染72 h后利用Western blot法检测转染前后Sema4D蛋白表达的变化;采用四甲基偶氮唑蓝显色法观察转染后细胞生长变化;Transwell迁移实验、划痕修复实验检测转染后人胰腺癌细胞迁移性的改变;小管形成实验观察转染后人胰腺癌细胞培养上清液对血管形成能力的影响。结果转染siRNA后,人胰腺癌细胞中Sema4D mRNA和Sema4D蛋白表达、胰腺癌细胞的生长速度均较阴性对照组和空白对照组下降(P<0.05);Transwell迁移实验和划痕修复实验显示,胰腺癌细胞穿膜细胞数和划痕修复率显著低于阴性对照组和空白对照组(P<0.05);小管形成实验显示,转染siRNA组、阴性对照组和空白对照组血管形成数目分别为0.50±0.02、1.45±0.60、1.37±0.52,组间比较差异有统计学意义(P<0.05)。结论 Sema4D-siRNA能够在胰腺癌细胞中引发RNA干扰效应,下调Sema4D基因表达,抑制胰腺癌细胞增殖,使胰腺癌细胞迁移能力明显下降,抑制血管生成。Objective To investigate the effects of semaphorin 4 D（ Sema4 D） on the proliferation,migration and angiogenic of human pancreatic carcinoma cells. Methods Sema4 D-siRNA was designed and synthesized and transfected into human pancreatic carcinoma cells. After 48 hours of transient infection,the changes of expression of Sema4 D mRNA before and after transfection were detected by reverse transcription-polymeruse chain reaction method. And after 72 hours of transient infection,the changes of expression of Sema4 D protein before and after transfection were detected by Western blot method. The changes of growth of the transfected cells were observed by methyl thiazolyl terazolium assay. Using transwell migration test and scratch repair test to detect the changes of migration ability of human pancreatic carcinoma cells after transfection. Using tubule formation assay to observe the effect of supernatant of pancreatic carcinoma cell cultures on angiogenesis after transfection.Results Compared with the negative control group and blank control group,the expression of Sema4 D mRNA and Sema4 D protein and the growth rate of pancreatic carcinoma cells decreased significantly（ P 0. 05）. In transwell migration test and scratch repair test,it was observed that Pancreatic cancer cells penetrating cell number and scratch repair rate were significantly lower than that in negative control group and blank control group（ P 0. 05）. Tubule formation assay showed that there were significant differences in angiogenesis numbers among siRNA transfection group（ 0. 5 ± 0. 02）,negative control group（ 1. 45 ±0. 60） and blank control group（ 1. 37 ± 0. 52）（ P 0. 05）. Conclusion Sema4 D-siRNA can induce RNA interference in pancreatic carcinoma cells and down-regulate the expression of Sema4 D gene,which can inhibit the proliferation of pancreatic carcinoma cells,significantly reduce the migration ability of pancreatic carcinoma cells and inhibit angiogenesis.

关 键 词：RNA干扰 轴突诱导因子4D 胰腺癌细胞 增殖和迁移能力 血管生成 

分 类 号：R735.9[医药卫生—肿瘤]