Inverse correlation of epidermal growth factor receptor messenger RNA induction and suppression of anchorage-independent growth by OSI-774, an epidermal growth factor receptor tyrosine kinase inhibitor, in glioblastoma multiforme cell lines
Inverse correlation of epidermal growth factor receptor messenger RNA induction and suppression of anchorage-independent growth by OSI-774, an epidermal growth factor receptor tyrosine kinase inhibitor, in glioblastoma multiforme cell lines
复制标题
DOI：
10.3171/jns.2004.100.3.0523
复制
发表时间：
2004-03-01
影响因子：
4.1
通讯作者：
Buchfelder, M
中科院分区：
文献类型：
--
作者：
Halatsch, ME;Gehrke, EE;Buchfelder, M
目的。表皮生长因子受体（EGFR）的定量和定性改变通常发生在人类的许多癌症中，包括恶性神经胶质瘤。当前研究的目的是评估 OSI-774（一种新型 EGFR 酪氨酸激酶抑制剂）对九种多形性胶质母细胞瘤 (GBM) 细胞系的分子和细胞作用。方法。分别使用半定量逆转录聚合酶链式反应、免疫细胞化学分析、库尔特计数、软琼脂克隆和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记/荧光激活细胞分选来检查OSI-774对EGFR信使(m)RNA和蛋白质的表达、增殖、不依赖锚定的生长和凋亡的影响。所有 p53 基因均进行了完整的双向测序。OSI-774 对锚定非依赖性生长的抑制与相对血清饥饿期间 EGFR mRNA 的诱导呈负相关 (r = -0.74)，并且与 p53 状态无关。总体而言，OSI-774 对贴壁依赖性生长的抑制作用比对增殖的抑制作用强得多。 OSI-774 诱导的细胞凋亡程度与 GBM 细胞系的增殖和贴壁依赖性生长呈正相关（r 分别为 0.75 和 0.79）。在源自继发性 GBM 的单细胞系中，暴露于大于或等于 1 mumol/L 的浓度会导致增殖研究期间大量的净细胞损失。结论。 EGFR mRNA 的诱导可能构成抵消 OSI-774 对 GBM 细胞贴壁依赖性生长的抑制作用的细胞机制。相比之下，GBM 细胞系中 EGFR（mRNA 和蛋白质）的基线表达水平与其对 OSI-774 的生物反应之间没有建立显着的相关性。 OSI-774 在更恶性的 GBM 表型中诱导更大的（p53 独立的）细胞凋亡，并且可能是一种有前途的继发性 GBM 治疗剂。
Object. Quantitative and qualitative alterations in the epidermal growth factor receptor (EGFR) commonly occur in many cancers in humans, including malignant gliomas. The aim of the current study was to evaluate molecular and cellular effects of OSI-774, a novel EGFR tyrosine kinase inhibitor, on nine glioblastoma multiforme (GBM) cell lines.Methods. The effects of OSI-774 on expression of EGFR messenger (m)RNA and protein, proliferation, anchorage-independent growth, and apoptosis were examined using semiquantitative reverse transcription-polymerase chain reaction, immunocytochemical analysis, Coulter counting, soft agar cloning, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling/fluorescence-activated cell sorting, respectively. All p53 genes were completely and bidirectionally sequenced.Suppression of anchorage-independent growth by OSI-774 was inversely correlated to the induction of EGFR mRNA during relative serum starvation (r = -0.74) and was unrelated to p53 status. Overall, suppression of anchorage-independent growth was a considerably stronger effect of OSI-774 than inhibition of proliferation. The extent of OSI-774-induced apoptosis positively correlated with both proliferation and anchorage-independent growth of GBM cell lines (r = 0.75 and 0.79, respectively). In a single cell line derived from a secondary GBM, exposure to concentrations of greater than or equal to 1 mumol/L resulted in a substantial net cell loss during proliferation studies.Conclusions. The induction of EGFR mRNA may constitute a cellular mechanism to counteract the inhibitory effect of OSI-774 on the anchorage-independent growth of GBM cells. In contrast, no considerable correlation could be established between baseline expression levels of EGFR (both mRNA and protein) in GBM cell lines and their biological response to OSI-774. The OSI-774 induced greater (p53-independent) apoptosis in more malignant GBM phenotypes and may be a promising therapeutic agent against secondary GBM.