Down-regulation of MT1-MMP expression suppresses tumor cell invasion in metastatic human SW626 ovarian cancer cells.
Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) is a key enzyme involved in degradation of extracellular matrix (ECM) and various surface-associated proteins that control cell growth, differentiation and survival, plays crucial roles in molecular carcinogenesis, tumor cell growth, invasion, and angiogenesis. We tested the inhibitory effect of antisense MT1-MMP on the ability of metastatic human ovarian carcinoma cell line SW626 in proliferation and invasion. RT-PCR was used to amplify MT1-MMP cDNA fragments with two different restriction sites at its 5'-end. Antisense MT1-MMP cloned in eukaryotic expression vector pMMP14as was transfected into SW626 cells. MT1-MMP protein expression, activities of MMP-2 and MMP-9, changes of cell proliferation, and cell invasion ability were detected by Western blot, optimized gelatin zymography, MTT assay and matrigel in vitro invasion assay, respectively. After 48 h transfection, decreased expression of endogenous MT1-MMP protein was detected in pMMP14as-transfected SW626 cells and showed significantly lower proliferation level when compared with control cells. The activation of proMMP-2 was inhibited markedly, and the mean percentage of invasive cells was 63.30+/-5.80% in pMMP14as-transfected cells, which was less than that (97.60+/-7.50%) in control cells (P<0.05). Both cell proliferation and invasion in SW626 cells were inhibited effectively by antisense MT1-MMP transfection, suggesting that MT1-MMP may be a proper target molecule for anti-invasion therapy for human ovarian cancers.
Wu
Mingfu
M
Cancer Biology Research Center, Tongji Hospital, Tongji Medical School, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, Hubei 430030, P.R. China.
Xu
Gang
G
Xi
Ling
L
Wei
Junchen
J
Song
Anping
A
Han
Zhiqiang
Z
Zhou
Jianfeng
J
Wang
Shixuan
S
Zhu
Tao
T
Zhang
Arli
A
Lu
Yunping
Y
Ma
Ding
D
D015153:Blotting, Western; D049109:Cell Proliferation; D015536:Down-Regulation; D005260:Female; D006801:Humans; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D053504:Matrix Metalloproteinases, Membrane-Associated; D009361:Neoplasm Invasiveness / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000201:enzymology* / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D020782:Matrix Metalloproteinases; D053504:Matrix Metalloproteinases, Membrane-Associated