Lysophospholipids Induce Matrix Metalloproteinases (MMPs) Expression in Human Endothelial Cells

水解磷酸脂對內皮細胞蛋白質水解酵素作用之研究

吳雯婷(Wen-Ting Wu) ； 李心予(Hsin-Yu Lee)

《Acta Zoologica Taiwanica》 14卷2期 (2004/10) Pp. 107-108

https://doi.org/10.6576/AZT.2004.14.(2).9

摘要

Lysophosphatidic Acid (LPA)與Sphingosine-1-phosphate (S1P)是形態與構造十分簡單的小分子水解磷酸脂(lysophospholipid, LPLs)，對細胞的生存、複製、增生及分化都扮演很重要的角色。LPA和S1P會經由和細胞表面特異接受器(Edgreceptors)之接合來達成對細胞的作用，進而活化各種中下游之細胞基質，以達到傳遞訊息的目的。Matrix metalloproteinases (MMPs)是一群zinc-dependent的蛋白質水解酵素，可分解許多細胞外間質(extracellular matrix)，參與在組織重建、胚胎發育、傷口修復等過程中，且會增加癌細胞侵入的能力，其中MMP-2是一個很重要的成員。在本研究中，使用RT-PCR，西方墨點法和substrate gel等實驗方法，探討水解磷酸脂對內皮細胞MMP-2表現的調控，發現包括在mRNA，蛋白質和酵素活性等層次皆受到調控。也觀察到S1P能調控MMP的天然抑制物，Tissue inhibitor of metalloproteiases的表現。因此推論水解磷酸脂可以經過對MMP-2表現的調控，造成細胞遷移或侵入能力增加。

關鍵字

水解磷酸脂 ； Lysophosphatidic Acid LPA ； 內皮細胞 ； Sphingosine-1-phosphate S1P

並列摘要

Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are both low molecular weight lysophospholipids (LPLs) that can promote cell proliferation, migration, and invasion via interaction with the endothelial differentiation gene (Edg) family of specific G protein-coupled receptors. Matrix metalloproteinases (MMPs) are a family of zinc-dependent enzymes, which are involved in degradation of extracellular matrix, and play critical roles in endothelial cell migration and matrix remodeling during wound healing and angiogenic process, etc. Among these MMPs, MMP-2 is an important molecule that can trigger the invasion of tumor cells. By RT-PCR, Western blotting, and gelatin zymography assays, we showed that LPA and S1P enhanced MMP-2 expression in mRNA, protein levels and enzymatic activity in concentration- and time-dependent manner. Next we investigated the signal transduction pathway of LPLs induced MMP-2 expression, which are involved in MEK/ERK-, NF-κB-, and calcium influx-dependent pathway. Furthermore, we use the chemotaxis chamber to investigate LPA and S1P effects on cell invasion, and the results showed that endothelial cell invasion was significantly induced by these LPLs. The induction can be prevented by pre-incubation with GM6001, a chemically synthesize MMP inhibitor. Interestingly, we found that S1P also regulated the nature inhibitors of MMPs, Tissue inhibitor of metalloproteiases (TIMPs) expression, including TIMP-2 and TIMP-3. These observations suggest that LPA and S1P may play key roles in endothelial cell invasion through regulating the expression of MMP-2.

並列關鍵字

lysophospholipids LPLs ； Lysophosphatidic Acid LPA ； endothelial cell ； Sphingosine-1-phosphate S1P