骨桥蛋白与乳腺癌细胞体外形成血管生成拟态的关系  

Relationship between Osteopontin and Vasculogenic Mimicry in Breast Cancer Cells in Vitro

作　　者：朱白鹭[1] 朱伟锋[1] 唐荣[2] 揭克敏[1] 

机构地区：[1]南昌大学医学院生物化学与分子生物学教研室,南昌330006 [2]抚州市第一人民医院检验科,江西抚州344000

出　　处：《南昌大学学报（医学版）》2012年第12期13-18,26,共7页Journal of Nanchang University：Medical Sciences

基　　金：江西省自然科学基金(00540036)

摘　　要：目的探讨骨桥蛋白(OPN)在乳腺癌细胞血管生成拟态(VM)的形成中的作用,及其与MMP-9和血管内皮生长因子(VEGF)的关系。方法实验分为转染组(miR-opn2组)和3个对照组[正常对照组(control组)、转染试剂对照组(LipLTX组)、阴性质粒对照组(miR-neg组)],分别转染MDA-MB-231细胞株30h后,收集细胞进行体外三维培养,24h后行PAS染色,显微镜下观察各组VM的数量,提取各实验组细胞,采用RT-PCR、WesternBlot测定OPN、MMP-9和VEGF的mRNA和蛋白质的表达。结果 miR-opn2组的MDA-MB-231细胞株形成VM的数量,OPN、MMP-9和VEGF的mRNA和蛋白表达水平较3个对照组的明显减少(P<0.05),3个对照组组间相比差异无统计学意义(P>0.05)。结论体外三维培养模式中人乳腺癌细胞株MDA-MB-231能够形成典型VM现象,靶向沉默OPN蛋白能够有效抑制体外三维培养模式中MDA-MB-231形成VM的数量,可能与其下调了VM相关蛋白MMP-9和VEGF的表达有关。Objective To investigate the effect of osteopontin(OPN) on the formation of vasculogenic mimicry(VM) in breast cancer cells,and to explore the relationship of OPN to MMP-9 and vascular endothelial growth factor(VEGF).Methods MDA-MB-231 cells were divided into four groups:normal control group(control group),transfection reagent control group(LipLTX group),negative vector control group(miR-neg group) and recombinant vector group(miR-opn2 group).After transfection for 30 hours,cells were collected and three-dimensionally cultured for 24 hours.The number of VM was detected under a microscope before and after PAS staining.The mRNA and protein expression of OPN,MMP-9 and VEGF were determined by RT-PCR and Western blot.Results Compared with miR-opn2 group,the number of VM and the mRNA and protein expression of OPN,MMP-9 and VEGF significantly increased in other three groups(P<0.05).There were no obvious differences among control group,LipLTX group and miR4-neg group(P>0.05).Conclusion Typical VM exists in three-dimensional culture of MDA-MB-231 human breast cancer cells.Targeted silencing of OPN protein can inhibit the formation of VM through reducing the expression of MMP-9 and VEGF.

关 键 词：乳腺癌 骨桥蛋白 血管生成拟态 体外三维培养 MMP-9 血管内皮生长因子 

分 类 号：R737.9[医药卫生—肿瘤]