[Inhibitory effect of MT1-MMP antisense nucleotide on invasion of human highly metastatic ovarian cancer cell line SW626].
OBJECTIVE
Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) is a newly discovered enzyme, which plays a key role in tumor metastasis. This study was to observe inhibitory effect of MT1-MMP antisense nucleotide on proliferation and invasive potential of human highly metastatic ovarian carcinoma cell line SW626.


METHODS
Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify MT1-MMP cDNA fragments with 2 different restriction sites at its 5c-end. RT-PCR products were cloned into plasmid pcDNA3.1 in antisense direction. The recombinant pMMP14as was transfected into SW626 cells. Changes of cell proliferation, MT1-MMP protein expression, activities of MMP-2 and MMP-9, and cell invasion ability were detected by MTT assay, Western blot, optimized gelatin zymography, and matrigel in vitro invasion assay, respectively.


RESULTS
Antisense MT1-MMP eukaryotic expression vector pMMP14as was constructed successfully. After 48-h transfection with pMMP14as, proliferation of pMMP14as-transfected SW626 cells was significantly lower than that of control cells. Compared with control cells, the expression of endogenous MT1-MMP protein in pMMP14as-transfected cells was decreased with a inhibition rate of 65.8%. The activation of proMMP-2 was remarkably inhibited, and the mean invasive cell percentage was (63.3+/-5.8)% in pMMP14as-transfected cells, which was far less than (97.6+/-7.5)% in control cells (P< 0.05).


CONCLUSIONS
Both cell proliferation and invasive potential of SW626 cells were inhibited effectively by antisense MT1-MMP, suggesting that MT1-MMP may be a proper molecular target of anti-invasion therapy for human ovarian cancer.
Wu
Ming-Fu
MF
Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430 030, P.R. China.
Liao
Guo-Ning
GN
Jia
Ping
P
Xi
Ling
L
Lu
Yun-Ping
YP
Ma
Ding
D
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics; D009361:Neoplasm Invasiveness; D016376:Oligonucleotides, Antisense; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2