Mmp 1 sirna

For the stable transfection of anti-miR-361-5p, anti-miR-361-5p sequence were amplified from miRZip-361-5p construct (System Biosciences) and cloned into pSilencer4.1 system. BC cells were then transfected with the pSliencer vector containing the antisense sequence of miR-361-5p. The cells were selected by puromycin after 48 h transfection and then diluted. MiR-361-5p mimics, miR-control, FGFR1 siRNA, MMP-1 siRNA or siRNA negative control were purchased from Genepharma (China). FGFR1 and MMP-1 cDNA ORF Clone were purchased from Origene (Origene Technology). Transient transfections were performed by using Lipofectamine 2000 (Invitrogen) following the manufacturer’s protocol. Cells were kept in medium containing 2% FBS for 48 h and then harvested and used.

Knockdown of PINK1, MMP1, NLRP3, and HSF1 was performed by reverse transfection of PINK1 siRNA (GenePharma, Shanghai, China), MMP1 siRNA (GenePharma), NLRP3 siRNA (GenePharma), and HSF1 siRNA (GenePharma). RBM3-overexpression plasmid pGV141-RBM3 was purchased from GeneChem (Shanghai, China). The cells were placed into the chambers of a 2-D clinostat 24 h after being transfected with the abovementioned siRNA or plasmid by using Lipofectamine 2000 (11668-019, Invitrogen, Waltham, Massachusetts, United States).