小鼠前列腺癌微环境中血管内皮生长因子对树突状细胞分化的影响  

Effect of VEGF on differentiation of dendritic cells in microenvironment of murine prostate cancer

作　　者：张蔚[1] 白文坤[1] 寿文德[1] 胡兵[1] 

机构地区：[1]上海交通大学附属第六人民医院超声医学科,上海超声医学研究所,上海200233

出　　处：《肿瘤》2015年第9期990-996,共7页Tumor

基　　金：国家自然科学基金资助项目(编号:81401421)~~

摘　　要：目的 :探讨小鼠前列腺癌微环境中树突状细胞的分化与血管内皮生长因子(vascular endothelial growth factor,VEGF)表达之间的关系。方法 :在BALB/c小鼠的骨髓腔冲洗液中加入细胞因子进行培养,获得树突状细胞(dentritic cells,DCs),应用FCM法检测其表面CD11c和CD83的表达情况。实时荧光定量PCR检测小鼠前列腺癌RM-1细胞中VEGF m RNA的表达,并将携带有VEGF抑制序列的重组载体质粒VEGF sh RNA-GV248转染至RM-1细胞中,筛选出阳性细胞VEGF-free RM-1。将DCs与RM-1细胞或VEGF-free RM-1细胞共培养3 d后,应用FCM法检测DCs表面CD11c和CD83的表达情况。结果 :骨髓腔冲洗液中加入细胞因子培养5 d后,其表面CD11c和CD83的阳性率分别为75.0%和14.6%,说明已获得DCs且部分已分化成熟。RM-1细胞中存在VEGF m RNA的表达。共培养组(DCs与RM-1细胞共培养)和抑制组(DCs与VEGF-free RM-1细胞共培养)DCs的CD11c和CD83阳性率均低于对照组(未进行共培养的DCs)(P值均<0.05),抑制组DCs的CD83阳性率高于共培养组(P<0.05)。结论 :前列腺癌微环境中DCs和成熟DCs的分化均受到抑制,而下调VEGF的表达可以促进成熟DCs的分化。Objective： To investigate the relationship of differentiation status of dendritic cells （DCs） and expression of vascular endothelial growth factor （VEGF） in microenvironment of murine prostate cancer. Methods： The bone marrow was extracted from BALB/c mice and co-cultured with cytokines to stimulate DCs. The expressions of CD11c and CD83 on the surface of DCs were detected by FCM.The expression of VEGF mRNA of murine prostate cancer RM-1 cells was detected by real- time fluorescent quantitative-PCR. The recombination vector VEGF shRNA-GV248 plasmid containing VEGF inhibitor sequence was transfected into RM-1 cells to screen out VEGF-free RM-1 cells. The expressions of CDI lc and CD83 on the surface of DCs after co-culture with RM-1 cells or VEGF-free RM-1 cells were detected by FCMResults： The positive expression rates of CD1]c and CD83 on the surface of DCs in bone marrow after co-culture with cytokines for 5 days were 75.0% and 14.6%, respectively, indicating that DCs including some mature cells were successfully stimulated. VEGF mRNA was expressed in RM-1 cells. The positive expression rates of CD11c and CD83 on the surface of DCs in co-culture group （DCs co-cultured with RM-1 cells） and suppression group （DCs co-cultured with VEGF-free RM-1 cells） were lower than that in the control group （DCs without any co-culture） （P 〈 0.05）. The positive expression rate of CD83 in the suppression group was higher than that in the co-culture group （P 〈 0.05）. Conclusion： The differentiation of DCs and mature DCs in microenvironment of murine prostate cancer was inhibited, while the down-regulation of VEGF expression can improve the differentiation of mature DCs.

关 键 词：前列腺肿瘤 树突状细胞 血管内皮生长因子类 

分 类 号：R737.25[医药卫生—肿瘤]