BMP9对共培养体系中人乳腺癌MDA-MB-231细胞迁移和侵袭的影响及其机制探讨  

Effect of BMP9 on migration and invasion of human breast cancer MDA-MB-231 cells in co-culture system and its mechanism

作　　者：王林[1] 段雯婷 王维[3] WANG Lin;DUAN Wenting;WANG Wei(Department of Clinical Laboratory,Xi′an NO.1 Hospital,Xi′an,Shaanxi 710002,China;Department of Cardiology,Xi′an NO.1 Hospital,Xi′an,Shaanxi 710002,China;Department of Clinical Laboratory,Xi′an Children Hospital,Xi′an,Shaanxi 710003,China)

机构地区：[1]陕西省西安市第一医院检验科,陕西西安710002 [2]陕西省西安市第一医院心血管内科,陕西西安710002 [3]陕西省西安市儿童医院检验科,陕西西安710003

出　　处：《国际检验医学杂志》2021年第23期2871-2875,共5页International Journal of Laboratory Medicine

基　　金：陕西省重点研发计划(2017SF-119)。

摘　　要：目的采用体外细胞共培养技术模拟骨微环境,探讨骨形态发生蛋白9(BMP9)对人乳腺癌MDA-MB-231细胞迁移和侵袭的影响,并探讨其机制。方法骨髓基质细胞HS-5与MDA-MB-231细胞在Transwell共培养体系中分别加入磷酸盐缓冲液(对照组)和重组BMP9蛋白(100 ng/mL)(实验组),培养2 d后,细胞划痕实验检测MDA-MB-231细胞迁移能力的改变;Transwell细胞侵袭实验检测MDA-MB-231细胞侵袭能力的改变;采用Western blot及明胶酶谱法分别检测MDA-MB-231细胞基质金属蛋白酶(MMP)9、MMP2蛋白表达及相对活性的改变;采用Western blot检测HS-5和MDA-MB-231两种细胞中基质细胞衍生因子-1(SDF-1)及MDA-MB-231细胞中趋化因子受体4(CXCR4)和细胞信号通路P-Akt表达的变化。结果在共培养体系中加入BMP9后,划痕实验结果显示,对照组和实验组MDA-MB-231细胞平均愈合率分别为(84.28±2.54)%、(44.49±4.86)%;Transwell细胞侵袭实验结果显示,对照组和实验组MDA-MB-231细胞穿膜细胞数分别为481±22、121±11,BMP9对共培养体系中MDA-MB-231细胞的迁移和侵袭有明显抑制作用(P<0.05)。实验组MDA-MB-231细胞MMP9、MMP2的蛋白表达分别为0.10±0.01、0.11±0.03,较对照组的0.79±0.03、0.82±0.05明显下调,差异均有统计学意义(P<0.05),实验组MMP9、MMP2相对活性分别为0.14±0.03、0.12±0.02,较对照组的0.90±0.07、1.04±0.14均明显下调,差异均有统计学意义(P<0.05);实验组MDA-MB-231细胞中CXCR4受体蛋白及其配体SDF-1的蛋白表达分别为0.08±0.01、0.06±0.02,与对照组的0.07±0.01、0.05±0.01比较,差异均无统计学意义(P>0.05);实验组P-Akt蛋白表达(0.09±0.01)较对照组(0.48±0.07)明显降低,差异有统计学意义(P<0.05);实验组HS-5细胞中SDF-1蛋白表达(0.14±0.01)较对照组(0.42±0.06)明显降低,差异有统计学意义(P<0.05)。结论在模拟骨微环境的共培养体系中,BMP9可能通过SDF-1/CXCR4-PI3K信号通路抑制人乳腺癌MDA-MB-231细胞的迁移和�Objective Using in vitro cell co-culture technology to simulate the bone microenvironment,to explore the effect of bone morphogenetic protein 9(BMP9)on the migration and invasion of human breast cancer MDA-MB-231 cells,and to explore its mechanism.Methods Bone marrow stromal cells HS-5 and MDA-MB-231 cells were added phosphate buffer(control group)and recombinant BMP9 protein(100 ng/mL)(experimental group)in the Transwell co-culture system,respectively.After 2 days of culture,the cell scratch test detected the change of MDA-MB-231 cell migration ability;Transwell cell invasion test detected the change of MDA-MB-231 cell invasion ability;Western blot and gelatin zymography method were used to detect the expression and relative activity of matrix metalloproteinase(MMP)9 and MMP2 protein in MDA-MB-231 cells respectively;Western blot was used to detect the expression change of stromal cell-derived factor-1(SDF-1)in HS-5 and MDA-MB-231 cells and the chemokine receptor 4(CXCR4)and cell signaling pathway P-Akt in MDA-MB-231 cells.Results After adding BMP9 to the co-culture system,the scratch test results showed that the average healing rate of MDA-MB-231 cells in control group and experimental group were(84.28±2.54)%and(44.49±4.86)%,respectively.The results of Transwell cell invasion test showed that the number of transmembrane cells of MDA-MB-231 cells in control group and experimental group were 481±22 and 121±11 respectively.BMP9 significantly inhibited the migration and invasion of MDA-MB-231 cells in the co-culture system(P<0.05).The expressions of MMP9 and MMP2 protein in MDA-MB-231 cells in experimental group were 0.10±0.01 and 0.11±0.03,respectively,which were significantly down-regulated compared with 0.79±0.03 and 0.82±0.05 in control group,and the differences were statistically significant(P<0.05).The relative activities of MMP9 and MMP2 in the experimental group were 0.14±0.03 and 0.12±0.02,respectively,which were significantly down-regulated compared with 0.90±0.07 and 1.04±0.14 in the control g

关 键 词：骨形态发生蛋白9 乳腺癌 骨微环境 迁移 侵袭 

分 类 号：R737.9[医药卫生—肿瘤] R446.1[医药卫生—临床医学]