BCSC-1基因异位表达对MDA-MB-231侵袭能力影响及其机制探讨  

Influence on invasion ability of MDA-MB-231 by ectopic expression of BCSC-1 gene and its mechanism

作　　者：邸大琳[1] 陈蕾[2] 王丽娜[1] 王洪伟[1] 魏兵[1] 王会东[3] 鞠吉雨[1] 

机构地区：[1]潍坊医学院免疫学教研室,山东潍坊261053 [2]潍坊医学院附属医院血液科,山东潍坊261031 [3]潍坊市人民医院乳腺外科,山东潍坊261041

出　　处：《中华肿瘤防治杂志》2014年第6期415-419,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省自然科学基金(2009CM019);山东省教育厅资助项目(2007WZ30);潍坊市卫生局科研计划(2012065)

摘　　要：目的:利用已构建的乳腺癌候选抑制蛋白-1(breast cancer suppressor candidate 1,BCSC-1)基因真核表达质粒转染人乳腺癌细胞株MDA-MB-231,观察BCSC-1基因异位表达对MDA-MB-231细胞体外侵袭能力的影响,初步探讨其影响机制。方法:利用脂质体将pcDNA3.1/v5-HisB-BCSC-1和空质粒pcDNA3.1/v5-HisB转染野生型MDA-MB-231细胞,以转染空质粒pcDNA3.1/v5-HisB的MDA-MB-231细胞为对照组,野生型MDA-MB-231细胞为空白对照组,转染后经G418加压筛选获得稳定BCSC-1表达细胞株。细胞划痕和Transwell细胞侵袭实验分析转染前后细胞侵袭能力的改变,实时荧光定量PCR分析转染前后基因的变化,细胞免疫组化分析转染前后BCSC-1以及细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)的变化。结果:转染后成功获得BCSC-1基因异位高表达稳定细胞株。细胞划痕实验显示,转基因组细胞平均迁移距离为(105.33±13.61)μm,较对照组(225±18.02)μm和对照组(236±16.46)μm明显降低,F=60.50,P=0.002。细胞侵袭实验显示,转基因组24h平均穿膜细胞数为34.67±2.52,较空白对照组56.66±2.08和对照组63.33±4.16明显降低,F=72.33,P=0.005。实时定量荧光PCR结果显示,转基因组BCSC-1表达水平为32.66±2.51,较对照组2.33±1.52和空白对照组明显升高,F=333.12,P=0.001;转基因组ICAM-1表达水平为28.67±3.79,较对照组3.33±1.53和空白对照组明显升高,F=127.14,P=0.003。细胞免疫组化结果显示,转染后BCSC-1以及ICAM-1表达水平明显增高。结论:BCSC-1基因的异位表达对MDA-MB-231细胞体外转移和侵袭能力有明显的抑制作用,这种抑制作用可能与ICAM-1表达增高有关。OBJECTIVE： To transfect human breast cancer cell MDA-MB-231 with BCSC-1 （Breast cancer suppres- sor candidate 1）gene eukaryotic expression vector, study the influence on invasion ability of MDA-MB-231 cells by ectopic expression of BCSC-1 gene in vitro and explore its mechanisms. METHODS： pcDNA3.1/v5-HisB-BCSC-1 and pcDNA3.1/ v5-HisB were transfeeted into wild type MDA-MB-231 cells by liposomes, pcDNA3. 1/v5-HisB transfected cells group were the control group,the wild type MDA-MB-231 cells were the blank controls. The stable cell line expressing BCSC-1 protein was successfully established. Cell scratch and transwell methods were used to detect the invasion ability,real-time fluorescence quantitative PCR and immunohistochemistry method were used to detect the expressions of BCSC-1 and ICAM-1 （Intercellular cell adhesion molecule 1）. RESULTS： The stable celi line expressing BCSC-1 protein was successful- ly established. Cell scratch assay showed that the average migration distance of blank control group, control group andtransgenic group were （225±18.02） μm, （236± 16.46） μm and （105.33± 13.61） μm respectively. The migration ability of transgenic group was significantly lower than that of the other two groups （F= 60.50, P=0. 002）. Cell invasion experi- ment showed that the average number of transwell cells of bank control group, control group and transgenic group were 56.66±2.08,63.33±4.16 and 34.67±2.52 respectively,the invasive ability of transgenic group was lower significantly than that of the other two groups （F= 72.33, P= 0. 005）. Real-time fluorescence PCR showed that BCSC-1 expression level were 2.33±1.52 and 32.66±2.51 in control group and transgenic group respectively compared with the blank con- trol group （F=333.12,P=0. 001）. ICAM-1 expression level were 3.33±1.53 and 28.67±3.79 respectively,the BCSC-1 and ICAM-lexpression levels in transgenic group were higher significantly compared with the other two groups （F= 127. 14,P=0. 003）. Immunohistochemist

关 键 词：乳腺肿瘤 BCSC-1基因 基因表达 MDA-MB-231细胞 

分 类 号：R737.9[医药卫生—肿瘤]