The FVIIa-tissue factor complex induces the expression of MMP7 in LOVO cells in vitro.
OBJECTIVE
The extracellular interactions of plasma clotting factor VIIa (FVIIa) with tissue factor (TF) on the cell surface trigger intracellular signaling events involved in multiple physiological processes. TF expression is related to the metastatic potential of tumor cells and is a significant risk factor in the development of hepatic metastases in patients with colorectal cancer. At present, it is unclear how the interaction between TF and FVIIa influences the development of metastasis in colon cancer.


METHODS
We used a stable LOVO cell line derived from colorectal adenocarcinoma for our model Western blot analysis, Northern blot analysis, polymerase chain reaction, and RNA inference (RNAi), and the Dual-Luciferase Reporter Assay System technology were utilized to determine if MMP7 can be up-regulated by the VIIa/TF complex.


RESULTS
Northern blot analysis confirmed that the plasma clotting factor FVIIa/TF complex resulted in a marked increase in MMP7 expression in a time- and dose-dependent manner via the p38 pathway in vitro. The proximal promoter of the human MMP7 gene was cloned into a luciferase reporter construction (MMP7.luc1592). Upon treatment with FVIIa, reporter activity in LOVO cells was increased by 2.5-fold. TF RNAi almost completely abolished FVIIa-mediated MMP7.luc induction. Deletion constructs from MMP7.luc1592 further defined an active promoter region.


CONCLUSIONS
Taken together, these data provide evidence that expression of MMP7 in colon cancer may be regulated by FVIIa and TF at the transcriptional level. MMP7 may act as a downstream mediator of FVIIa/TF signal transduction to facilitate the development of metastasis in colon cancer.
Laboratory of Colon Cancer, First Hospital of Peking University, Peking University, No. 8, Xishiku Street, West District, Beijing 100034, China.
Jian-Quan
JQ
Zhang
Yuan-Lian
YL
Wan
Yu-Cun
YC
Liu
Xin
X
Wang
Jian-Qiang
JQ
Tang
Tao
T
Wu
Jing
J
Zhu
Yi-Sheng
YS
Pan
D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015942:Factor VIIa / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D020783:Matrix Metalloproteinase 7 / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011725:Pyridines / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000235:genetics*; D014158:Transcription, Genetic; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D004791:Enzyme Inhibitors; D007093:Imidazoles; D011725:Pyridines; D012334:RNA, Neoplasm; D048051:p38 Mitogen-Activated Protein Kinases; D015942:Factor VIIa; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7; C093642:SB 203580