The Effect Of Nimesulide On Proliferation, Apoptosis And Expression Of ETS-1, MMP-1 On Human Primary Cultured Meningioma Cells

Objective:Meningiomas, which are the tumors that arise from the arachnoid cap cells, comprise approximately 13–25 % of all tumors of the central nervous system. Clinical evidence supports the concept that meningioma formation may be associated with previous head trauma. The evidence suggests that meningioma formation may occur in the setting of chronic inflammation triggered by trauma. Cox-2 is the rate-limiting enzyme in the synthesis of prostaglandins from arachidonic acid. Prostaglandins have angiogenic, cell-proliferative, and antiapoptotic properties. The Ets-1 transcription factor and mmp-1 play an important role in various physiologic and pathologic processes requiring extracellular remodeling such as tumor invasion, angiogenesis and metastasis. Nimesulide, a selective cyclooxygenase-2 (COX-2) inhibitor, has the effects of antiproliferative and proapoptotic property on tumor cells. The aim of the research is to find the possibility of Nimesulide in treatment of human meningioma by means of studying cell proliferation, apoptosis and the expression of ETS-1 and MMP-1 in the human meningioma.Methods:Establishing the short-term primary cell culture method of human meningioma first, and then detects the expressions of EMA and Vinmitin protein by immunohistochemistry (IHC) respectively. Meningioma cells were treated with various concentrations of nimesulide(75～600μmol /L).The effect of nimesulide on cell proliferation was measured by MTT and apoptosis was determined by ao-eb ,tunnel and DNA ladder analysis. The expression of ETS-1 and MMP-1 were measured by western blot.Results:1. Successfully established the short-term primary cell culture of 13 human meningioma cases, including the condition of short-term cell culture, and the methods of passage, Cryopreservation, cell revitalization, identification of tumor cell.2. In the course of culture, we can found that cells in medium containing nimesulide were different from control culture, outline of cells were dim, refraction was weak, and more vacuole and granule can be seen in cells. Some cells separated from the wall. This change is dose-depended. Whereas the control cells were more transparent, outline of cells were clear, refraction was strong.3. Immunohistochemistry showed a high expression of Ets-1 and MMP-1 in tumor cells. In the cells, Ets-1 and MMP-1 expression were reduced after 24 and 48 hours by nimesulide.4. The MTT results suggested that nimesulide at concentrations of 75,150,300,600μmol/ L,had inhibitory effects on the proliferation of meningioma cells .The higher the concentration of nimesulide was, the stronger the cytotoxic effect reached, which suggested obvious dose-dependent manner of nimesulide .5. the nimesulide has the ability to induce apoptosis of the meningioma cells in a dose- and time- effective manner。Conclusion:It suggests that nimesulide has the capabilities of inhibiting proliferation and inducing apoptosis in meningioma cells in vitro,the effects is in a dose and time dependt-manner,its cytostatic mechanism may be associated with reducing the expression of ETS-1 and MMP-1.