沉默CD147基因对人乳腺癌MDA-MB-231细胞的影响  

Impact of siRNA-mediated down-regulation of CD147 on human breast cancer cells

作　　者：李振乾[1] 李道明[1] 李江伟[1] 黄培[1] 秦慧[1] 

机构地区：[1]郑州大学第一附属医院病理科河南省肿瘤病理重点实验室,450052

出　　处：《中华病理学杂志》2015年第10期734-738,共5页Chinese Journal of Pathology

摘　　要：目的探讨沉默CD147基因对人乳腺癌MDA—MB-231细胞生长、增殖、运动能力的影响。方法构建CD147小干扰RNA（siRNA）慢病毒载体并转染各实验组MDA—MB-231细胞，采用免疫细胞化学ABC法检测沉默前后MDA—MB-231细胞中CD147、基质金属蛋白酶（MMP）-2、金属蛋白酶类组织抑制剂（TIMP）-2蛋白的表达水平；采用即时定量PCR和WesternBlot技术检测CD147、MMP-2、TIMP-2的mRNA和蛋白表达水平改变；采用CCK_8法和划痕实验分别检测细胞的增殖和迁移能力；建立裸鼠皮下移植瘤模型，观察CD147siRNA对裸鼠瘤体质量和体积的影响。结果转染MDA．MB-231细胞后，CD147、MMP-2、TIMP-2蛋白呈弱阳性或阴性表达，明显低于对照组（P〈0．01）。转染72h时，CD147基因下调平均效率为96．03％±0．84％，MMP一2基因下调平均效率48．64％±0．35％，mRNA表达均明显下降（P〈0．05），而TIMP-2mRNA表达水平与对照组相比差异无统计学意义（P〉0．05）。转染第2天后，MDA-MB-231细胞生长明显受到抑制（P〈0．05）。转染24h后MDA—MB-231细胞平均迁移距离为（0．64±0．12）mm，而空白对照组细胞平均迁移距离为（4．69±0．85）mm，迁移速度减小。沉默CD147基因后，裸鼠瘤体体积和瘤体质量均下降（P〈0．05），平均瘤体质量为（1．85±0．98）g，移植瘤的生长受到抑制（P〈0．05）。结论CD147可以影响MDA—MB-231细胞中的MMP-2／TIMP-2平衡，沉默CD147基因可以抑制MDA—MB-231细胞的增殖和迁移及裸鼠移植瘤的生长。Objective To investigate the influence of siRNA-mediated down-regulation of CD147 on growth, proliferation and movement of human breast cancer cell line MDA-MB-231. Methods The protein expression of CD147, MMP-2 and TIMP-2 of the MDA-MB-231 cells were analyzed by ABC. Lentiviral expression vector of CD147 gene was constructed and transfected into MDA-MB-231 cells. RT-PCR and Western blot were used to detect the mRNA and protein level changes of CD147 genes to identify the optimal time point, followed by detection of changes of mRNA and protein expression of MMP-2 and TIMP-2 genes. CCK-8 reagent method and cell scratch test were used to detect the proliferation and migration change of MDA-MB-231 cells. The nude mouse model of breast cancer by hypodermic injection with MDA-MB-231 cells was established to document the effect of CD147 siRNA on the tumor transplants. Results After transfection of lentiviral expression vector of CD147 gene, protein of CD147, MMP-2 and TIMP-2 were weakly or negative expressed, significantly weaker than those of control group（ P 〈 0. 01 ）. After 72 hours of transfection, average down-regulation rate of CD147 and MMP-2 were 96. 03% ± 0. 84% and 96. 03% ± 0. 84%, respectively. Both CD147 mRNA and MMP-2 mRNA expression were down-regulated（P 〈0. 05）, while TIMP-2 mRNA expression showed no significant deference （ P 〉 O. 05 ）. No less than 2 days after transfection, cell growth of MDA-MB-231 cell line was found significantly inhibited （ P 〈 0. 05 ） . After 24 hours of tranfection, average migration distance of MDA-MB-231 cell line and control group were（0.64 ±0. 12）ram and （4. 69 ±0. 85）mm, respectively, which indicated a lower migrate speed. Down regulation of CD147 led to reduction of volume and mass of nude mouses. The growth of the carcinoma transplant was inhibited upon siRNA-mediated down-regulation of CD147 （ P 〈 0. 05 ）, with an average tumor mass of （ 1.85 ±0. 98） g and both reduction of tumor size and tumor mass. Conclusions CD147 may al

关 键 词：乳腺肿瘤 细胞系 肿瘤 抗原 CD147 基质金属蛋白酶2 金属蛋白酶类组织 抑制剂 

分 类 号：R737.9[医药卫生—肿瘤]