摘要
目的探讨细胞外基质蛋白1(extracellular matrix protein 1,ECM1)在肿瘤中的生物学功能。方法构建ECM1-pEGFP-N2真核表达载体,利用脂质体介导的转染技术转染MCF-7细胞,药物G418筛选稳定转染细胞株,荧光显微镜检测报告基因表达产物EGFP,免疫组化检测ECM1蛋白表达。经细胞粘附实验、体外侵袭力实验比较转染前后细胞侵袭力的变化;用MTT比色法分析ECM1对MCF-7和人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖的影响。结果成功构建了ECM1-pEGFP-N2真核表达载体,并在MCF-7中稳定表达;转染ECM1后的MCF-7细胞的细胞形态、粘附性、侵袭力和增殖能力无明显变化;MTT比色法检测HUVEC增殖结果示培养液组、空载体转染上清组、ECM1转染上清组HU-VEC D570值分别为0.89±0.06,0.92±0.09和1.39±0.10,各组间差异具有显著统计学意义(p<0.01)。结论ECM1对乳腺癌细胞株MCF-7的生物学特性在体外未见明显影响,但能显著促进血管内皮细胞体外增殖。
Objective To investigate the biological effect of extracellular matrix protein 1(ECM1) on breast tumor cell MCF-7 line and HUVEC.Methods ECM1-pEGFP-N2 was transfected with Lipofectamine to MCF-7 cells;MCF-7 cells harboring ECM1 genes were selected by G418 pressure and the expression of ECM1-pEGFP-N2 was determined by GFP fluorescence and anti-ECM1 immunohistochemistry.The adhesion and migration of MCF-7 cells were measured by cell counting and a modified Boyden chamber assay;The proliferation of MCF-7 and HUVEC was measured by MTT colorimetry assay analysis.Results ECM1 Eukaryotic Expression Vector ECM1-pEGFP-N2 was successfully constructed and the ECM1 gene was successfully expressed in MCF-7 cells.The morphology,adhesion,migration and proliferation of ECM1 transfected MCF-7 cells were not significantly different with those untreated and mock transfected MCF-7 cells.Proliferation of HUVEC(1.39±0.10) in ECM1 transfected group was higher than those untreated(0.89±0.06) and mock transfected group(0.92±0.09)(p<0.01).Conclusion ECM1 has no effect on MCF-7 in vitro,but it can significantly promote the proliferation of HUVEC.
出处
《医学检验与临床》
2008年第4期25-28,共4页
Medical Laboratory Science and Clinics
基金
国家自然科学基金项目(30080027)
上海市基础研究重大项目(02JC14005)
上海市松江区科学技术攻关项目(06KG27)资助
关键词
细胞外基质蛋白1
肿瘤
真核表达载体
MCF-7
人脐静脉内皮细胞
Extracellular matrix protein 1(ECM1)
Tumor
Eukaryotic expression vector
MCF-7
Human umbilical vein endothelial cells(HUVEC)
