Suppression of basement membrane type IV collagen degradation and cell invasion in human melanoma cells expressing an antisense RNA for MMP-1

被引:55
作者
Durko, M
Navab, R
Shibata, HR
Brodt, P
机构
[1] MCGILL UNIV,DEPT SURG,DIV SURG RES,MONTREAL,PQ H3A 1A1,CANADA
[2] ROYAL VICTORIA HOSP,MONTREAL,PQ H3A 1A1,CANADA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH | 1997年 / 1356卷 / 03期
关键词
invasion; interstitial collagenase; metalloproteinase; metastasis;
D O I
10.1016/S0167-4889(97)00004-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
During progression from benign nevus to vertical growth phase melanoma, melanocytes acquire the ability to invade into the dermis. This process requires rupture of the basal lamina and dissolution of dermal type I collagen. Metastases-derived human melanoma MIM cells have an invasive ability in vitro which is dependent on metalloproteinases. In the present study we analysed the role of type I collagenase (MMP-1) in melanoma invasion using MIM cells in which the constitutive expression of MMP-1 was suppressed by stable transfection with a plasmid vector expressing a 777 bp antisense fragment of MMP-1 genomic DNA. Two clones were isolated in which MMP-1 mRNA expression was blocked by 90-96% with a corresponding loss in protein synthesis. In their morphological appearance and growth rate in vitro these cells were indistinguishable from wild type cells or control cells transfected with the same vector expressing the MMP-1 fragment in the sense orientation. Their mRNA and protein levels for type IV collagenase (MMP-2) were unchanged as assessed by Northern and Western blot analyses and by gelatin zymography. However, when the invasive ability of the cells was measured, we found that in addition to type I collagen, invasion through type IV collagen and a reconstituted, type IV collagen-containing basement membrane (Matrigel) were also significantly inhibited as compared to normal or sense-transfected cells. The results indicate that despite the presence of functional MMP-2, degradation of type IV collagen matrices by the melanoma cells was dependent on expression of MMP-1. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:271 / 280
页数:10
相关论文
共 44 条
[1]   ROLE OF PLASMINOGEN ACTIVATORS, METALLOPROTEINASES AND THE TISSUE INHIBITOR OF METALLOPROTEINASE-1 IN THE METASTATIC PROCESS OF HUMAN SALIVARY-GLAND ADENOCARCINOMA CELLS [J].
AZUMA, M ;
TAMATANI, T ;
FUKUI, K ;
YOSHIDA, H ;
KAMOGASHIRA, T ;
OGINO, K ;
SUZUKI, T ;
SATO, M .
INTERNATIONAL JOURNAL OF CANCER, 1993, 54 (04) :669-676
[2]  
BERNHARD EJ, 1990, CANCER RES, V50, P3872
[3]   DIFFERENCES IN THE REPERTOIRES OF BASEMENT-MEMBRANE DEGRADING ENZYMES IN 2 CARCINOMA SUBLINES WITH DISTINCT PATTERNS OF SITE-SELECTIVE METASTASIS [J].
BRODT, P ;
REICH, R ;
MOROZ, LA ;
CHAMBERS, AF .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1139 (1-2) :77-83
[4]  
BROWN PD, 1990, CANCER RES, V50, P6184
[5]  
CHEN JM, 1991, J BIOL CHEM, V266, P5113
[6]  
COLLIER IE, 1988, J BIOL CHEM, V263, P10711
[7]  
COLLIER IE, 1988, J BIOL CHEM, V263, P6579
[8]   HUMAN PROGELATINASE-A CAN BE ACTIVATED BY MATRILYSIN [J].
CRABBE, T ;
SMITH, B ;
OCONNELL, J ;
DOCHERTY, A .
FEBS LETTERS, 1994, 345 (01) :14-16
[9]   RECIPROCATED MATRIX METALLOPROTEINASE ACTIVATION - A PROCESS PERFORMED BY INTERSTITIAL COLLAGENASE AND PROGELATINASE-A [J].
CRABBE, T ;
OCONNELL, JP ;
SMITH, BJ ;
DOCHERTY, AJP .
BIOCHEMISTRY, 1994, 33 (48) :14419-14425
[10]  
DIPASQUALE G, 1986, P SOC EXP BIOL MED, V183, P262