细胞外基质金属蛋白酶诱导因子对脑胶质瘤血管生成拟态体外形成的影响及其机制

Effect and mechanism of extracellular matrix metalloproteinase inducer on the vasculogenic mimic- ry formation of glioma

目的探讨细胞外基质金属蛋白酶诱导因子（EMMPRIN）对脑胶质瘤血管生成拟态体外形成的影响及其机制。方法应用三维培养人脑胶质瘤细胞株U87，构建EMMPRIN基因干扰及过表达载体，转染人胶质瘤细胞株U87，实时定量聚合酶链反应（Real—timePCR）技术和Westernblot技术检测EMMPRIN对人脑胶质瘤细胞株U87血管生成拟态体外形成的影响和酶联免疫吸附试验（ELISA）技术检测基质金属蛋白酶（MMP）-2、MMP-9蛋白表达变化。结果三维培养时，恶性胶质瘤细胞株U87能够在Matrigel基质胶上形成类似于内皮细胞的环状结构，EMMPRIN过表达组（HeRNA—EMMPRIN）、空白对照组（Control—EMMPRIN）、干扰组（siRNA—EMMPRIN）各组细胞株U87的血管生成拟态平均管状结构的总结点数分别为26．93±3．59、17．33±2．41和5．40±2．22，形成能力逐渐减弱及细胞上清MMP-2／MMP-9蛋白表达逐渐减低，且相互之间差异有统计学意义（P〈0．01）。结论EMMPRIN可能通过调控MMP-2／MMP-9蛋白的表达来影响脑胶质瘤血管生成拟态的形成。

Objective To investigate the effect and mechanism of extracellular matrix metallopro- teinase inducer （EMMPRIN） on the vasculogenic mimicry formation of glioma. Methods Three - dimen- tional culture was performed on the glioma cell line U87. EMMPRIN gene was intervened and overexpressed. U87 ceils were transfected. Real -time quantitative polymerase chain reaction （Real -time PCR） and West- ern blotting were used to observe the effect of EMMPRIN on the formation of vasculogenic mimicry in vitro. ELISA was used to detect the expressions of matrix metalloproteinase （MMP） -2/MMP- 9 protein. Results U87 - MG cells could form a kind of tubular structures analogous to the endothelial cells on Matrigel. The kind of tubular structures in HeRNA - EMMPRIN group, control - EMMPRIN group and siRNA - EMMPRIN group was 26. 93 ± 3.59, 17.33 ± 2. 41 and 5.40 ± 2. 22 respectively, showing the ability of vasculogenic mimicry formation in U87 - MG cells and the expression of MMP - 2/MMP - 9 proteins were gradually weakened （P 〈 0. 01 ）. Conclusion EMMPRIN can influence the ability of vasculogenic mimicry formation in glioma probably by regulating the expression of MMP -2/MMP -9 proteins.