siRNA mediated inhibition of MMP-1 reduces invasive potential of a human chondrosarcoma cell line.
Matrix metalloproteinases (MMPs) play a crucial role in tumor cell invasion and metastasis. Expression of MMP-1 has been reported as a prognostic predictor of recurrence in human chondrosarcoma, and studies using human chondrosarcoma cell lines indicate that MMP-1 expression levels correlate with in vitro invasiveness. These observations suggest that MMP-1 activity has a central role in cell egress from the primary tumor at an early step in the metastatic cascade. In this study, siRNA was used to investigate whether knock down of the MMP-1 gene could be used to inhibit invasiveness in a human chondrosarcoma cell line. The inhibitory effect of siRNA on endogenous MMP-1 gene expression and protein synthesis was demonstrated via RT-PCR, Northern blotting, Western blotting, collagenase activity assay, and an in vitro cell migration assay. The siRNA inhibited MMP-1 expression specifically, since it did not affect the expression of endogenous glyceraldehyde phosphate dehydrogenase (GAPDH) nor other collagenases. Most importantly, the siRNA mediated reduction in MMP-1 expression correlated with a decreased ability of chondrosarcoma cells to invade a Type I collagen matrix. The reduction of invasive behavior demonstrated by human chondrosarcoma cells transfected with MMP-1 siRNA and the specificity of this inhibition supports the hypothesis that this metalloproteinase molecule is involved in initiation of chondrosarcoma metastasis.
Jiang
Xiaoling
X
Department of Orthopedic Research, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA.
Dutton
Charyl M
CM
Qi
Wen-Ning
WN
Block
Joel A
JA
Garamszegi
Nandor
N
Scully
Sean P
SP
D045744:Cell Line, Tumor; D002465:Cell Movement; D002813:Chondrosarcoma / Q000378:metabolism* / Q000473:pathology*; D024042:Collagen Type I / Q000378:metabolism; D017077:Culture Media, Conditioned; D018390:Gene Targeting; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*
D024042:Collagen Type I; D017077:Culture Media, Conditioned; D034741:RNA, Small Interfering; D020781:Matrix Metalloproteinase 1