巨噬细胞集落刺激因子和粒细胞-巨噬细胞集落刺激因子对小鼠乳腺癌4T1细胞迁移及血管内皮生长因子A表达的影响

Effect of M-CSF and GM-CSF on migration and VEGF-A expression of breast cancer cell line 4T1

目的 探讨巨噬细胞集落刺激因子（M-CSF）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）对乳腺癌4T1细胞迁移及血管内皮生长因子（VEGF）-A表达的影响.方法 分别用5、10 ng/ml M-CSF和GM-CSF处理小鼠乳腺癌4T1细胞株,采用实时荧光定量PCR方法检测4T1细胞中细胞因子VEGF-AmRNA表达量的变化.划痕实验和Transwell实验检测用5 ng/ml M-CSF、5 ng/ml GM-CSF和10 ng/mlVEGF-A对4T1细胞迁移和侵袭能力的影响.结果 经5、10ng/ml M-CSF分别处理4T1细胞后VEGF-AmRNA在12h和24 h的相对表达量分别为17.81±2.49和17.48±5.43、5.15±2.59和5.45±4.28;经5、10 ng/ml GM-CSF分别处理4T1细胞后VEGF-A mRNA在12h和24 h的相对表达量分别为9.77±2.39和7.61±2.80、6.53±2.41和6.30±2.89.与无细胞因子处理的对照组相比,除10 ng/ml GM-CSF处理4T1细胞24 h组VEGF-A mRNA表达水平差异无统计学意义（P＞0.05）外,其他各组VEGF-A mRNA表达水平差异均有统计学意义（均P＜ 0.05）,处理细胞12h时VEGF-A的表达较24 h时更明显（均P＜ 0.01）.划痕实验显示M-CSF和GM-CSF可促进4T1细胞的迁移,但VEGF-A对4T1细胞的迁移无影响.Transwell实验发现,用M-CSF、GM-CSF和VEGF-A处理的实验组中4T1细胞的穿膜数量多于无细胞因子处理的对照组（均P＜ 0.05）.结论 M-CSF和GM-CSF可促进小鼠乳腺癌4T1细胞迁移及VEGF-A的表达.

Objective To investigate the effect of M-CSF and GM-CSF on migration and expression of VEGF-A in breast cancer cell line 4T1.Methods Real-time PCR was used to detect VEGF-A mRNA expression in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF or GM-CSF.Ability of migration and metastasis of 4T1 cells were analyzed by scratch and Transwell assays.Results The relative expression of VEGF-A mRNA at 12 h and 24 h in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF were 17.81±2.49 and 17.48± 5.43,5.15±2.59 and 5.45±4.28,respectively,while those treated by GM-CSF were 9.77±2.39 and 7.61±2.80,6.53±2.41 and 6.30±2.89,respectively.M-CSF and GM-CSF can promote the expression of VEGF-A in 4T1 cells （P 〈 0.05）.The relative expression of VEGF-A was higher in 4T1 cells treated for 12 h than that for 24 h （P 〈 0.01）.M-CSF,GM-CSF and VEGF-A can promote metastasis of 4T1 cells （all P 〈 0.05）,whereas no gross migration of 4T1 cells was showed by VEGF-A treatment.Conclusion M-CSF and GM-CSF can promote the migration and expression of VEGF-A in breast cancer cell line 4T1.