摘要
目的探讨肺癌细胞分泌的血管内皮生长因子(VEGF)促进肺转移的受体相关机制。方法应用噻唑蓝(MTT)与酶联免疫吸附(ELISA)方法测定9种肺癌细胞系的增殖状况和培养上清液中VEGF水平,筛选出差异表达VEGF且增殖无明显差异的两株细胞系。将两株细胞分别接种于SCID小鼠背部观察肿瘤生长,肺癌细胞经尾静脉注射建立肺转移模型,采用HE染色和免疫荧光实验检测肺转移瘤及血管密度。应用两种VEGFR中和抗体(MF-1和DC101)腹腔注射治疗肺转移小鼠,观察肺转移瘤数改变。结果 9种肺癌细胞系分泌VEGF水平不同,其中最高的是A549(182.7 ng/mL),最低的是SPCA1(13.39 ng/mL),A549细胞和SPCA1细胞的增殖差异无统计学意义(P>0.05)。A549细胞在小鼠背部形成的肿瘤体积明显大于SPCA1细胞,肿瘤组织内新生血管明显增多。A549细胞形成肺转移瘤数为SPCA1细胞的2.3倍。抗VEGFR-1治疗使肺转移瘤数明显减少,而抗VEGFR-2治疗后差异无统计学意义(P>0.05)。结论肺癌细胞分泌的VEGF促进肿瘤生长、转移和肿瘤血管生成,VEGF通过VEGFR1通路促进肺转移。
Objective To investigate the mechanism by which vascular endothelial growth factor （VEGF） promotes lung metastasis. Methods VEGF levels of nine human lung carcinoma cell lines were examined with ELISA and the proliferation was tested by MTT. Two cell lines with distinctive VEGF expression and similar proliferation were subcutaneously injected to the back of SCID mice or intravenously injected by the tail vein to build the metastasis model. The volume of tumors on back of mice were measured every 3 days till sacrificed. Pulmonary metastasis and vascular density were verified by HE staining and CD31 immunofluorescence. Neutralizing antibodies specific for mouse VEGFR1 （MF1） and VEGFR2 （DC101） were administrated to verify which receptor was involved in VEGF induced pulmonary metastases. Results ELISA showed that VEGF levels of nine human lung carcinoma cell lines were in a range of 13.39-182.7ng/mL. A549 cell with high VEGF expression （182.7ng/mL） and SPCA1 with low VEGF expression （13.39ng/mL） were chosen to build mouse model. The proliferation of A549 and SPCA1 had no significant difference. A549 cell formed much larger tumor on the back of SCID mice than SPCA1. Angiogenesis in A549 formed tumor was significantly increased than SPCA1 formed tumor （P〈0.01）. In lung metastasis model, number of metastatic lesions in lung tissues found in A549 mice was 2.3 folds compared with SPCA1. After treatment with anti-mouse VEGFR1 monoclonal antibody, the total number of metastatic tumors induced by A549 decreased from 54 to 13, while anti-VEGFR2 treatment had no significant difference in metastasis numbers（P〉0.05）. Conclusion VEGF secreted by lung cancer cells could promote tumor growth and tumor angiogenesis. VEGF promotes pulmonary metastasis by VEGFR1 pathways. The blockage of VEGFR1 induced metastasis may provide a novel approach for prevention and treatment of tumor metastasis.
出处
《山东大学学报（医学版）》
CAS
北大核心
2013年第11期25-29,共5页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金(81272351)
山东省自然科学基金(2R2012HM020)
山东省科技发展计划(2012G0021826)
关键词
血管内皮生长因子
转移
血管内皮生长因子受体
肺肿瘤
动物模型
Vascular endothelial growth factor
Metastasis
Vascular endothelial growth factor receptor
Lung carcinoma
Animal model
