SPARC affects glioma cell growth differently when grown on brain ECM proteins in vitro under standard versus reduced-serum stress conditions.
SPARC affects glioma cell growth differently when grown on brain ECM proteins in vitro under standard versus reduced-serum stress conditions.
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在标准和低血清应激条件下，SPARC 在体外脑 ECM 蛋白上生长时，对神经胶质瘤细胞生长的影响不同。
DOI：
10.1093/neuonc/5.4.244
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发表时间：
2003
期刊：
影响因子：
15.9
通讯作者：
Rempel,SandraA
中科院分区：
文献类型：
--
作者：
Vadlamuri,SatyaV;Media,Joe;Sankey,SteadmanS;Nakeff,Alexander;Divine,George;Rempel,SandraA
当使用亲本 U87T2 和 U87T2 衍生的 SPARC 转染克隆进行体外和体内评估时，富含半胱氨酸的酸性分泌蛋白 (SPARC) 对 U87 神经胶质瘤细胞增殖具有抑制作用。由于 SPARC 与细胞外基质 (ECM) 蛋白相互作用，我们在标准（10% 胎牛血清 [FBS]）和降低（0.1% FBS）血清应激条件下，在存在和不存在 ECM 蛋白的情况下，研究了 SPARC 分泌对体外生长的神经胶质瘤细胞的增殖、形态和细胞密度的影响。在标准条件下，MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑）生长曲线、形态学和蛋白质印迹分析表明，SPARC 对生长具有抑制和双相作用，且不受 ECM 的严重调节。 24 小时观察到的 SPARC 诱导的形态变化并未因 ECM 的存在而改变。在降低血清应激条件下，蛋白质印迹、形态学和流式细胞术分析表明，当细胞在塑料上生长时，SPARC 诱导的抑制生长效应被消除。然而，观察到 ECM 特异性的生长变化，其中一些与分泌的 SPARC 水平相关。这些结果表明 SPARC 和 ECM 对增殖的不同影响取决于培养条件。由于在标准条件下获得的结果与我们的体内观察结果一致，因此我们得出结论，SPARC抑制增殖的能力在更大程度上受SPARC水平的调节，并且这种抑制作用不受任何所检查的ECM的存在的影响。
Secreted protein acidic and rich in cysteine (SPARC) has a suppressive effect on U87 glioma cell proliferation when assessed in vitro and in vivo using parental U87T2 and U87T2-derived SPARC-transfected clones. Since SPARC interacts with extracellular matrix (ECM) proteins, we examined the effect of SPARC secretion on proliferation, morphology, and cell density of glioma cells grown in vitro, in the absence and presence of ECM proteins under standard (10% fetal bovine serum [FBS]) and reduced (0.1% FBS) serum stress conditions. Under standard conditions, MTT (3-(4,5-cimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide) growth curves, morphology, and Western blot analyses demonstrated that SPARC had a suppressive and biphasic effect on growth that was not grossly modulated by the ECMs. The SPARC-induced changes in morphology observed at 24 h were not altered by the presence of ECMs. Under reduced-serum stress conditions, Western blot, morphological, and flow cytometric analyses indicated that the SPARC-induced suppressive growth effects were eliminated when the cells were grown on plastic. However, ECM-specific changes in growth were observed, some of which correlated with secreted SPARC levels. These results indicate that the differential effects of SPARC and ECMs on proliferation are dependent on culture conditions. Since the results obtained under standard conditions agree with our in vivo observations, we conclude that the ability of SPARC to suppress proliferation is regulated to a greater degree by the level of SPARC and that this suppressive effect is not influenced by the presence of any of the ECMs examined.