[Regulation of promatrilysin expression by tissue factor/actived coagulation factor VII complex in LoVo colon cancer cell line].
OBJECTIVE
To assess the expression of Promatrilysin in LoVo colon cancer cell by FVIIa stimulation,and to investigate the effect of MAPKs signal transduction pathway on up-regulation of Promatrilysin.


METHODS
(1) The expression of ProMMP-7 was detected by Western blot at different time points (0,2,4,6,9,12 and 24 h) and with different doses of (0,0.1,1,5,10,25 and 100 nmol/L) FVIIa stimulation. The change of ProMMP-7 expression was observed with 5 mg/L tissue factor (TF) antibody prior to 100 nmol/L FVIIa. (2) The activation of MAPKs (ERK, p38, JNK) signaling pathways were assessed at different time points after being stimulated with 100 nmol/L FVIIa and the changes of ProMMP-7 expression were detected after the special signal pathway inhibitors (PD98059,SB203580,SP600125) were applied,respectively.


RESULTS
(1) The expression of ProMMP-7 in LoVo cells was up-regulated by FVII a in a time-effect dependent and dose-effect dependent manner,and markedly reached the peak level at h12, 5.5 folds that of the control group (P=0.006).The up-regulation of ProMMP-7 was completely inhibited by blockade with TF antibody. (2) A time-dependent phosphorylation of ERK1/2 and P38 in LoVo cells was induced with FVIIa incubation,reached the peak at min10,2.2 folds and 3.9 folds those of the control groups respectively, but not JNK. (3) The upregulation effect of ProMMP-7 was partially blocked after incubation of ERK1/2 inhibitors PD98059 and P38 inhibitors SB203580 prior to FVIIa, The expression of ProMMP-7 decreased by 32%+/-5% and 61%+/-10% respectively (P<0.05).whereas JNK inhibitors SP600125 did not have the effect.


CONCLUSIONS
FVIIa induces tissue factor-dependent up-regulation of ProMMP-7 in LoVo cells. ERK1/2 and p38 signal pathways are not only involved in TF/FVIIa mediated signaling,but also related to the upregulation of MMP-7 in LoVo cells.
Tang
Jian-qiang
JQ
Department of General Surgery, Peking University First Hospital, Beijing 100034, China.
Wan
Yuan-lian
YL
Liu
Yu-cun
YC
Rong
Long
L
Wang
Xin
X
Wu
Tao
T
Pan
Yi-sheng
YS
Zhu
Jing
J
D000873:Anthracenes / Q000494:pharmacology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism*; D005167:Factor VII / Q000494:pharmacology*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020783:Matrix Metalloproteinase 7 / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D013925:Thromboplastin / Q000494:pharmacology*
D000873:Anthracenes; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; D005419:Flavonoids; D007093:Imidazoles; D011725:Pyridines; C432165:pyrazolanthrone; D005167:Factor VII; D013925:Thromboplastin; D008666:Metalloendopeptidases; C099583:promatrilysin; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one