MCP-1／CCR2轴促进脐带间充质干细胞向肺癌组织归巢  

MCP-1/CCR2 axis promotes the homing of human umbilical cord mesenchymal stem cells to lung cancer tissues

作　　者：宋新苗[1] 颜次慧[1] 吕梦果 于文文[1] 张新伟[1,2] 任秀宝[1,2] 

机构地区：[1]天津医科大学肿瘤医院免疫室,国家肿瘤临床医学研究中心,天津市肿瘤免疫与生物治疗重点实验室,天津300060 [2]天津医科大学肿瘤医院生物治疗科,天津300060

出　　处：《中国肿瘤生物治疗杂志》2016年第1期17-23,共7页Chinese Journal of Cancer Biotherapy

基　　金：国家重点基础研究发展计划(973计划)资助项目(No.2012CB9333004);国家科技支撑计划资助项目(No.2015BAI12B12);国家自然科学基金资助项目(No.81401887);天津市科委应用基础基金资助项目(No.14JCQNJC11500,No.11JCYBJC13200);天津医科大学科学基金资助项目(No.2012KYM03)~~

摘　　要：目的:探讨单核细胞趋化蛋白-1(monocyte chemoatractant protein-1,MCP-1)/趋化因子(C-C模体)受体2[chemokine(C-C motif)receptor 2,CCR2)轴在人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUMSCs)向肺癌归巢中的作用。方法:采用组织块培养法从健康新生儿脐带组织中分离HUMSCs并鉴定,构建BALB/c裸鼠皮下移植瘤模型。体外应用Transwell趋化实验、体内应用IVIS Xenogen动物活体成像系统检测HUMSCs是否向肺癌归巢,ELISA法检测肺癌细胞A549培养上清中MCP-1分泌水平,转染shRNA敲低肺癌细胞内MCP-1的表达和用抑制剂RS504393抑制HUMSCs细胞表面的MCP-1受体CCR2后,体内外检测HUMSCs向肺癌归巢能力的改变。结果:成功分离得到HUMSCs并完成鉴定,成功建立BALB/c裸鼠皮下肺癌移植瘤模型。HUMSCs在体内外均能向肺癌归巢(P<0.01),肺癌细胞高表达MCP-1。成功构建稳定低表达MCP-1的肺癌A549细胞系,与对照组相比,shRNA1和shRNA2敲低组趋化HUMSCs的数目明显减少[(80.0±33.0)、(94.0±16.0)vs(167.0±41.0)个,均P<0.05],抑制HUMSCs细胞表面MCP-1受体CCR2后,体内外均显著抑制HUMSCs向肺癌的趋化能力(P<0.05)。结论:MCP-1/CCR2轴促进HUMSCs向肺癌归巢。Objective：To study the role of monocyte chemoatractant protein-I （MCP-1）/（chemokine [ C-C motif] receptor 2, CCR2） axis in human umbilical cord mesenchymal stem cells （HUMSCs） homing to lung cancer. Methods： ＂Tissue explant＂ method was used to isolate and identify HUMSCs from umbilical cord of healthy newborns. Subcutaneous lung cancer xenograft model was established in nude BALB/c mice. Transwell migration assay in vitro and IVIS Xenogen living imaging system in vivo were applied respectively to investigate the capability of HUMSCs homing to lung cancer. ELISA was utilized to detect the secretion of monocyte chemoatrractant protein-1 （ MCP-1 ） in lung cancer A549 cell culture supernatant. After knocking down the MCP-1 expression in lung cancer cells by transfecting shRNA and blocking MCP-1 receptor CCR2 on the surface of HUMSCs by inhibitor RS504393, the migration ability of HUMSCs was determined in vitro and in vivo. Results： HUMSCs were successfully isolated and identified. Subcutaneous lung cancer xenograft model was successfully established in nude BALB/c mice. HUMSCs migrated to lung cancer both in vivo and in vitro （ P 〈 0.01 ）. Lung cancer cells highly expressed MCP-1. Lung cancer A549 cell line with stable low expression of MCP-1 was success- fully constructed, compared with control group, number of migrating HUMSCs significantly decreased in shRNA1 and shRNA2 knock down groups （ [ 80.0 ± 33.0 ], [ 94.0 ± 16.0 ] vs [ 167.0 ±41.0 ], P 〈 0.05 ）. Inhibition of CCR2 （ re- ceptor of MCP-1 ） on HUMSCs greatly suppressed the tropism of HUMSCs to lung cancer in vivo and in vitro （ P 〈 0.05 ）. Conclusion： MCP-1/CCR2 axis promoted the migration of HUMSCs to lung cancer.

关 键 词：间充质干细胞 单核细胞趋化蛋白-1 趋化因子(C-C模式)受体2 肺癌 归巢 

分 类 号：R734.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]