Increased SPARC expression promotes U87 glioblastoma invasion in vitro
Increased SPARC expression promotes U87 glioblastoma invasion in vitro
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DOI：
10.1016/s0736-5748(99)00009-x
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发表时间：
1999-08-01
影响因子：
1.8
通讯作者：
Rempel, SA
中科院分区：
文献类型：
--
作者：
Golembieski, WA;Ge, S;Rempel, SA
我们最近的研究集中在识别在脑肿瘤进展早期表达的促进侵袭的基因。我们确定并表征了作为潜在候选物的α-半胱氨酸（分泌性酸性蛋白质，富含半胱氨酸）。为了确定增加的cDNA 3表达功能是否促进脑肿瘤侵袭，使用四环素关闭基因表达系统将cDNA 3转染到U87 MG胶质母细胞瘤细胞中。对亲本细胞系（U87 MG）、亲本反式激活因子转染的克隆（U87 T2）和三个选定的U87 T2-SPARC转染的克隆（A2 bi、A2 b2和C2 a4）进行内源性和转染的miR表达表征。与亲本或U87 T2细胞系相比，经SPARC转染的克隆证明：（1）形态学变化，（2）增加的由四环素类似物多西环素下调的β转录物和蛋白质丰度，（3）转染后的细胞核定位（与报道的在培养物中的正常细胞中的BMP 3的定位一致）和（4）改变的粘附和增加的侵袭，如通过球体对抗测定所评估的。这些数据表明，增加的U87胶质母细胞瘤表达有助于U87胶质母细胞瘤的体外侵袭，这些细胞系将作为有用的试剂来研究U87在体外和体内促进这种表型的机制。(C)1999年ISDN。由爱思唯尔科技有限公司出版。保留所有权利。
Our recent studies have focused on identifying invasion-promoting genes that are expressed early in brain tumor progression. We identified and characterized SPARC (secreted protein acidic and rich in cysteine) as a potential candidate. To determine whether increased SPARC expression function ally promotes brain tumor invasion, SPARC was transfected into U87MG glioblastoma cells using the tetracycline-off gene expression system. The parental cell line (U87MG), the parental transactivator-transfected clone (U87T2) and three selected U87T2-SPARC-transfected clones (A2bi, A2b2 and C2a4) were characterized for endogenous and transfected SPARC expression. In comparison to the parental or U87T2 cell lines, the SPARC-transfected clones demonstrated: (1) morphological changes, (2) increased SPARC transcript and protein abundances that were down-regulated by the tetracycline analog doxycycline, (3) perinuclear localization of the transfected SPARC (consistent with reported localization of SPARC in normal cells in culture) and (4) altered adhesion and increased invasion as assessed by the spheroid confrontation assay. These data indicate that increased SPARC expression contributes to U87 glioblastoma tumor invasion in vitro and that these cell lines will serve as useful reagents to investigate the mechanism(s) by which SPARC promotes this phenotype in vitro and in vivo. (C) 1999 ISDN. Published by Elsevier Science Ltd. All rights reserved.