摘要
目的:利用小干扰RNA技术沉默Bmi-1基因的表达,探讨其对人乳腺癌细胞株MDA-MB-231侵袭转移能力的影响。方法:设计并化学合成了针对Bmi-1的小干扰RNA,转染具有高转移性的MDA-MB-231细胞,利用qRT-PCR和Western blot分别检测Bmi-1基因mRNA和蛋白的表达水平。体外通过Transwell小室、Matrigel胶模型﹑流式细胞术,观察Bmi-1被沉默后对乳腺癌细胞株MDA-MB-231生物学行为的影响。结果:qRT-PCR和Western结果显示,Bmi-1-siRNA转染组Bmi-1基因表达水平被有效抑制。与对照组相比,Bmi-1-siRNA转染组的细胞侵袭转移能力明显下降,细胞周期发生明显变化,细胞的增殖受到明显抑制。结论:抑制Bmi-1基因的表达可明显抑制MDA-MB-231细胞的侵袭迁移能力。
Objective：To investigate the effect of small interference RNA-mediated silencing of the Bmi-1 gene on cell invasion and metastasis in human mammary carcinoma cell line MDA-MB-231. Methods： Chemically synthesized siRNA targeting the Bmi-I gene was transfected into MDA-MB-231 cells, which have high invasive and metastatic potential. The expression of Bmi-1 mRNA and protein was detected by quantitave Real-time PCR and Western blot, respectively. The effect of Bmi-1 knockdown on MDA-MB-231 cells migration and invasion was analyzed by Transwell migration assay and Matrigel invasion assay. Results： Transfeetion with Bmi-I siRNA significantly down-regulated the expression of Bmi-1 RNA and protein as compared with the control group. MDA-MB-231 cells transfection with Bmi-1 siRNA had lower levels of invasion and migration capacity than cells in the control group. Conclusion： siRNA- mediated silencing of the Bmi-I gene could significantly inhibit cell migration and invasion in human breast cancer cell line MDA-MB- 231.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2014年第4期475-479,485,共6页
Chinese Journal of Immunology
基金
教育部高等学校博士学科点专项科研基金(20125503110012)
教育部高等学校博士学科点新教师基金项目(20115503120007)
重庆市自然科学基金项目(cstc2011jjA10035)
