Construct The Eukaryotic Expression Vector Of Id1 MiRNA And Transfect Into Breast Caner Cell Line To Explore The MMP9 Expression

[Purpose]: To construct the miRNA eukaryotic expression vector of Id1 and transfect the MDA-MB-231 breast cancer cell line,and finally select the stabilized expressed cells lines. To explore the relationship between Id 1 and MMP 9 expression in breast cancer line MDA-MB-231 .[Method]: The miRNA array of Id1 gene was designed and insert into pcDNA6.2-GW/EmGFP-miR expression vector to construct the Id1 miRNA eukaryotic expression vector. The liposome infection protocol was used to transfect the MDA-MB-231 cell lines. Use Blasticidin to select stabilized expressed cell lines.the level of Id1 and MMP 9 was detected using RT-PCR methods.[Result]: The eukaryotic expression vector of Id1 miRNA was successfully constructed After Identifying by electrophoresis and measuring array, the MDA-MB-231 breast cancer cell line was tranfected. We acquire the stabilized expressed cell lines through 2W Blasticidin selection. As a result of RT-PCR Identification, the level of Id1 and MMP 9 in Id1 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.[Conclusion]:The experiment successfully construct the eukaryotic expression vector of Id1 miRNA and successfully transfect the MDA-MB-231 breast cancer cell line and finally select the stabilized expressed cells lines.It prove that Id1and MMP9 are closely related.