Expression and stability of c-sis mRNA in human glioblastoma cells.

Expression and stability of c-sis mRNA in human glioblastoma cells.
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人胶质母细胞瘤细胞中 c-sis mRNA 的表达和稳定性。

DOI:
10.1021/bi00415a051
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发表时间:
1988
期刊:
影响因子:
2.9
通讯作者:
Goldthwait,DA
Goldthwait,DA
中科院分区:
生物学3区
文献类型:
--
作者:
Press,RD;Samols,D;Goldthwait,DA

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凯斯西储大学生物化学系,克利夫兰,俄亥俄州44106 1987年10月1日接收; 1988年1月7日接收修订的Mandarin pt摘要:胶质母细胞瘤产生的血小板衍生生长因子样(PDGF样)物质可能参与正常细胞向肿瘤细胞的转化。在这个问题的调查中,我们已经研究了血小板衍生生长因子B链mRNA(c-sis mRNA)的一些特性的灵敏和定量RNA-RNA溶液杂交方法。在8个人胶质母细胞瘤细胞系中,有5个细胞中存在c-sis mRNA,在最高水平的细胞系中,每个细胞中约有4-10个分子。c-sis mRNA在两种胶质母细胞瘤细胞系中的半衰期分别为2.6和3.4 h,而在人脐静脉内皮(HUVE)和膀胱癌(T24)细胞中的半衰期分别为1.6和2.5 h。抑制蛋白质的合成没有产生显着的改变c-sis mRNA的半衰期在胶质母细胞瘤或HUVE细胞。因此,在放线菌酮存在下,c-sis mRNA 3 '端的富含AU的序列似乎不像在其他转录物中那样影响mRNA的稳定性。正常细胞和肿瘤细胞中c-sis mRNA半衰期的相似性表明,c-sis mRNA稳定性的调节不是检查的胶质母细胞瘤细胞系中肿瘤发生的主要因素。正常胶质细胞转化为胶质母细胞瘤的机制仍然知之甚少。然而,现在有一些数据表明,在神经胶质细胞肿瘤发生中涉及自分泌生长环(Sporn & Todaro,1980),由此生长因子受体被改变和/或生长因子本身被异常产生。例如,表皮生长因子(EGF)受体基因已显示在许多人胶质母细胞瘤中扩增或结构改变(Libermann等人,1985年)。由经胎盘乙基亚硝基脲(ENU)诱导的大鼠神经胶质母细胞瘤在c-er B-B-2(neu)转化基因的跨膜结构域中含有点突变(Bargmann等人,1986),其结构与EGF受体基因具有相当大的同源性(Schechter等,1984年)。用ENU类似处理的大鼠脑细胞的长期培养导致表型转化,其时间与c-sis原癌基因的表达相关(透镜等人,1986),编码血小板衍生生长因子(PDGF)的B链的基因。这些ENU-in-
Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106 Received October 1, 1987; Revised Manuscript Received January 7, 1988 abstract: The production of platelet-derived growth factor like (PDGF-like) material by glioblastomas may be involved in the conversion of normal cells to tumor cells. In an investigation of this problem, we have examined some of the properties of the platelet-derived growth factor B-chain mRNA (c-sis mRNA) by a sensitive and quantitative RNA-RNA solution hybridization method. In 5 out of 8 human glioblastoma cell lines, c-sis mRNA was present, and in the linewith the highest level, there were approximately 4-10 molecules per cell. The half-lives of the c-sis mRNA in two glioblastoma cell lines were 2.6 and 3.4 h, while in human umbilical vein endothelial (HUVE) and bladder carcinoma (T24) cells they were 1.6 and 2.5 h, respectively. Inhibiting protein synthesis produced no significant alteration of the c-sis mRNA half-lives in the glioblastoma or HUVE cells. The AU-rich sequence at the 3'end of the c-sis mRNA therefore does not appear to affect the mRNA stability in the presence of cycloheximide as it does in other transcripts. The similarity of the c-sis mRNA half-lives in normal and tumor cells suggests that regulation of stability of c-sis mRNA is not a major factor in tumorigenesis in the glioblastoma celllines examined.] V^ Iechanisms by which normalglial cells are converted to glioblastomas are still poorly understood. Some data are now available, however, suggesting the involvement of an autocrine growth loop (Sporn & Todaro, 1980) in glial cell tumori-genesis, whereby growth factor receptors are altered and/or growth factors themselves are abnormally produced. The epidermal growth factor (EGF) receptor gene, for instance, has been shown to be either amplified or structurally altered in a number of human glioblastomas (Libermann et al., 1985). Rat neuroglioblastomas induced by transplacental ethylnitrosourea (ENU) contain a point mutation in the trans-membrane domain of the c-erb-B-2 (neu) transforming gene (Bargmann et al., 1986), the structure of which has considerable homology to the EGF receptorgene (Schechter et al., 1984). Long-term culture of neonatalrat brain cells similarly treated with ENU results in phenotypic transformation, the timing of whichcorrelates with expression of the c-sis protooncogene (Lens et al., 1986), the gene encoding the B chain of platelet-derived growth factor (PDGF). These ENU-in-
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