Duffy抗原趋化因子受体抑制人乳腺癌裸鼠移植瘤的生长和肺转移  

Duffy antigen receptor for chemokines attenuates breast cancer growth and metastasis:an experiment with nude mice

作　　者：王杰 欧周罗 侯意枫 罗建民 陈奕[2] 周晋[2] 沈镇宙 丁健[2] 邵志敏 

机构地区：[1]复旦大学附属肿瘤医院乳腺外科复旦大学乳腺癌研究所复旦大学上海医学院肿瘤学系,上海200032 [2]中国科学院上海药物研究所

出　　处：《中华医学杂志》2005年第29期2033-2037,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30371580);国家杰出青年科学基金资助项目(30025015);上海市科委重点项目基金资助项目(03JC14019);上海市自然科学基金资助项目(04ZR14027)

摘　　要：目的探讨Duffy抗原趋化因子受体(DARC)对人乳腺癌细胞株成瘤性和肺转移的影响及其作用机制。方法建立DARC过表达的MDA-MB-435肺高转移性细胞株(MDA-MB-435HM-DARC),观察体外细胞生物学行为的改变,检测培养上清中DARC配体CXCL8(IL-8)和CCL2(MCP-1,JE)浓度;建立人乳腺癌BALB/c裸鼠原位移植瘤模型,观察肿瘤生长曲线与肺内转移灶,检测瘤内DARC、JE、CD34、基质金属蛋白酶-9(MMP-9)等的变化,并进行相关性分析。结果MDA-MB-435HM-DARC在体外的增殖及侵袭性未见明显改变,但培养上清中CXCL8、CCL2的浓度明显降低。体内实验显示MDA-MB-435HM-DARC移植瘤生长延迟而且体积缩小,肺内转移灶数目减少,同时瘤内CCL2蛋白水平、微血管密度(MVD)降低,MMP-9的产生也减少。结论DARC作为促血管生成趋化因子的清除槽,可抑制由趋化因子诱导的血管生成,从而抑制乳腺癌的生长和肺转移。Objective To study the effects of Duffy antigen receptor for chemokines （DARC） on the tumorigenesis and metastasis of breast cancer. Methods Human breast cancer cells of the line MDA-MB-435HM with a great potentiality of metastasis were cultured and then divided into 3 groups： MDA-MB-435HM-DARC cell group, to be transfected with pcDNA3/DARC FyB containing human DARC cDNA;MDA-MB-435HM-vect cell group, to be transfected with blank plasmid pcDNA3 ; and MDA-MB-435HM cell group without transfection. Female BALB/c nude mice were implanted with MDA-MB-435HM cells into the nipple. The size of the implanted cancer was measured once a week. The mice were killed 4, 6, and 8 weeks after the implantation respectively and their lungs were taken out to observe the number of metastatic tumor. Immunohistochemical staining was performed to calculate the microvascular density （MVD）. Western blotting was used to detect the matrix metalloproteinase （MMP）-9 protein expression of the lung tissues. ELISA was used to detect the concentrations of the 2 DARC ligands： CNCL8 and CCL2 in the supernatant of culture fluid of the 3 kinds of cells and the metastatic tumors. Results The expression levels of DARC mRNA and protein of the MDA-MB-435HM cells were 38% those of the MDA-MB-435 cells. The expression levels of CXCL8 and CCL2 of the MDA-MB-435HM-DARC cells were significantly lower than those of the MDA-MB-435HM-vect cells and MDA-MB-435HM cells （both P 〈0. 05）. All mice developed tumor with a tumorigenesis rate of 100%. However, the tumor occurred 17 days after implantation in those mice implanted with MDA-MB-435HM-DARC cells, significantly later then in those implanted with MDA-MB-435HM-vect cells and MDA-MB-435HM cells （9 and 10 days after implantation） the size of tumor in the former group being significantly smaller than those of the 2 latter groups （ both P 〈 0. 01 ）. The number of metastatic foci in the lung of the MDA-MB-435HM-DARC group was significantly less than those of the other 2 groups （both P

关 键 词：乳腺肿瘤 肿瘤转移 血管生成 DUFFY抗原趋化因子受体 

分 类 号：R737.9[医药卫生—肿瘤]