乳腺癌细胞分泌的血管内皮生长因子诱导肺预转移小生境的形成  

Vascular endothelial growth factor secreted by breast cancer cells plays a critical role in the formation of pre-metastatic niche in the mouse lung

作　　者：李冉冉[1] 袁冰[1] 张颖[1] 戴建建[1] 张鹏飞[1] 房菲菲 韩明勇[1] 

机构地区：[1]山东大学附属省立医院肿瘤中心,济南250021

出　　处：《中华肿瘤杂志》2016年第1期17-22,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金（81272351）

摘　　要：目的探讨乳腺癌细胞诱导骨髓来源细胞（BMDCs）肺预转移小生境的形成及其相关机制。方法将乳腺癌4T1细胞接种至雌性小鼠乳腺，建立乳腺癌小鼠实验动物模型。应用共聚焦显微镜观察BMDCs在肺组织中的聚集，应用小鼠细胞因子芯片检测小鼠血清及4T1细胞培养上清中细胞因子的表达水平，应用血管内皮生长因子（VEGF）连续腹腔注射处理小鼠，观察VEGF对肺预转移小生境形成的作用。结果接种4T1细胞第0天，荷瘤组和对照组小鼠肺组织中均无绿色荧光蛋白（GFP）标记的BMDCs聚集。接种4T1细胞后第7、14天，荷瘤组小鼠肺组织中GFP标记的BMDCs数目分别为（8．7±2．2）个／高倍镜视野和（48．8±3．2）个／高倍镜视野，对照组分别为（1．1±0．8）个／高倍镜视野和（3．1±1．7）个／高倍镜视野，差异均有统计学意义（均P〈0．05）。共聚焦显微镜可见1，1’-双十八烷-3，3，3’，3＇-四甲基吲哚羰花青．高氯酸盐标记的乳腺癌细胞多在BMDCs聚集处定植。荷瘤小鼠血清中VEGF、粒细胞-巨噬细胞集落刺激因子和白介素6水平均高于对照组，差异均有统计学意义（均P〈0．05）；4T1细胞上清中VEGF水平高于RPMI1640培养基上清，差异有统计学意义（P〈0．05）。VEGF组和对照组小鼠肺组织中BMDCs数目分别为（22．8±3．6）个／高倍镜视野和（3．1±0．4）个／高倍镜视野，差异有统计学意义（P〈0．05）。VEGF组小鼠肺组织中转移瘤数目为（36．8±5．4）个，对照组小鼠肺组织中转移瘤数目为（12．6±2．2）个，差异有统计学意义（P〈0．05）。结论原发乳腺癌细胞能够诱导肺预转移小生境形成，这一过程与VEGF密切相关。Objective To explore the formation of pre-metastatic niche in the mouse lung and to study the underlying molecular mechanisms whereby primary breast carcinoma-derived factors mediate recruitment of bone marrow-derived cells （BMDCs） and affect the formation of pre-metastatie lung environment before the arrival of tumor cells. Methods Mammary carcinoma 4T1 cells were inoculated into the mammary gland to construct mouse model of breast cancer. Confocal microscopy was used to detect the recruitment of BMDCs in the pre-metastatic lungs. The expression of factors in the mouse sera and 4T1 cell culture media was assayed using RayBio Custom mouse eytokine antibody array kit. The mice were injected daily with recombinant VEGF for 7 consecutive days to observe the effect of VEGF on BMDCs recruitment in the mouse lung. Results No BMDCs were observed in the lungs of control and 4Tl-tumor-bearing mice on day 0. On day 7 and 14, clusters of BMDCs observed in the lungs of 4Tl-tumor-bearing mice were 8.7±2.2/ objective field and 48.8±3.2/objective field, respectively, significantly higher than those in the control mice （ 1.1±0.8/objective field and 3.1 ± 1.7/objective field） （ P〈0.05 for both）. Confocal microscopic observation found that metastatic breast cancer cells preferentially facilitate BMDCs recruitment sites in the pre-metastatic mouse lungs. The levels of VEGF, GM-CSF, and IL-6 in the serum of 4Tl-tumor-bearing mice were significantly increased compared with those in the control group （P〈0.05 for all）. However, VEGF was detected only in the culture media of 4T1 cells. The amount of BMDCs in the mouse lung tissue was （ 22.8±3.6）/objective field in the ~EGF group and （3.1 ±0.4）/objective field in the control group （P〈0.05）. There were 36.8±5.4 metastatic foci in the lung tissue of VEGF group and 12.6±2.2 in the control group （P〈 0.05）. Conclusions The results of this study demonstrate that primary breast cancer cells can alter the lung microenvironment during the pre-metast

关 键 词：乳腺肿瘤 肿瘤转移 血管内皮生长因子类 肺 预转移小生境 

分 类 号：R737.9[医药卫生—肿瘤]