分化和DNA结合抑制因子1基因对MDA-MB-231乳腺癌细胞株MMP9表达的影响

Relationship between Idl gene expression and MMP9 in MDA-MB-231 breast cancer cell line

目的筛选成功转染Id1 miRNA真核表达载体乳腺癌MDA-MB-23 1细胞系,观察对乳腺癌细胞的对Id 1、MMP 9表达的影响。方法构建Id1miRNA真核表达载体,转染乳腺癌MDA-MB-231细胞株以杀稻瘟霉素筛选稳定表达的细胞株。经RT-PCR方法鉴定其对Id1、MMP9表达的影响。结果经过筛选,获得转染Id1 miRNA真核表达载体稳定表达的MDA-MB-231细胞系。经RT-PCR方法鉴定,结果表明转染Id1 miRNA真核表达载体的乳腺癌MDA-MB-231细胞株其Id 1、MMP 9表达水平低于对照组,且Id 1、MMP 9表达水平正相关。未转染组与阴性对照转染组无明显差异。结论本实验成功筛选出稳定表达Id1 miRNA的乳腺癌细胞株,证实Id1 miRNA真核表达载体构建成功干扰有效,并且Id1与mmp9表达相关,为将来进一步研究他们之间的生物学行为差异,或详细了解其分子作用机制奠定了基础。

Objective To Select the the stabilized expressed cells lines of miRNA eukaryotic expression vector of Id1.To explore the relationship between Id 1 and MMP 9 expression in breast cancer line MDAMB -231.Methods Use Blasticidin to select stabilized expressed cell lines of miRNA eukaryotic expression vector of Id1.the level of Id1 and MMP 9 was detected using RT-PCR methods.Results The stabilized expressed cell lines through 2W Blasticidin selection.As a result of RT-PCR identification,the level of Id1 and MMP 9 in Idl miRNA transfeced cell line is lower than that in the control groups.While there is no difference between the not transfected and negative control transfected cell line.