Department of Pathology, Dupuytren University Hospital, 2 Avenue Martin Luther King, F-87042 Limoges, France.
Int J Oncol. 2012 Apr;40(4):1142-52. doi: 10.3892/ijo.2011.1287. Epub 2011 Dec 8.
The epidermal growth factor receptor (EGFR) gene encodes four alternatively spliced mRNA, variants 1, 2, 3 and 4, respectively, encoding the whole isoform a (EGFR) and truncated isoforms b, c and d, all of which lack the receptor's intracellular domain. In addition, a mutant EGFRvIII differs from isoform a in a truncated extracellular domain. The expression pattern of these isoforms is unknown in adult diffuse gliomas. Thus, we investigated in 47 cases: i) EGFR protein expression by immunohistochemistry using an extracellular domain-recognizing antibody (Ext-Ab) and an intracellular domain specific one (Int-Ab), ii) mRNA expression of EGFRv1, -v2, -v3, -v4 and -vIII by RT-PCR and iii) EGFR amplification by fluorescent in situ hybridization. The relation of these data with histological criteria and patient outcome was studied. The immunostaining was stronger with the Ext-Ab than with the Int-Ab. EGFRv1, -v2, -v3 and -v4 mRNA expression were highly correlated. They were expressed in all tumors, with highest levels in glioblastomas. EGFRv1 strong levels and the presence of vIII mRNAs were more closely associated with Int-Ab staining. EGFR gene amplification concerned only glioblastomas and was associated with the presence of EGFRvIII and high levels of EGFRv2, -v3 and -v4 transcripts. A pejorative outcome was associated with: histology (glioblastomas), EGFR amplification, strong Int-Ab labeling and high levels of variant mRNAs. Our results indicated that the full-length EGFR and mutant EGFRvIII are not the sole EGFR isoform expressed in diffuse gliomas. This could explain discordant immunohistochemical results reported in the literature and may have therapeutic implications.
表皮生长因子受体(EGFR)基因编码四个选择性剪接的 mRNA,分别为变体 1、2、3 和 4,分别编码全长异构体 a(EGFR)和截短异构体 b、c 和 d,所有这些都缺乏受体的细胞内结构域。此外,突变型 EGFRvIII 在截断的细胞外结构域上与异构体 a 不同。这些异构体在成人弥漫性神经胶质瘤中的表达模式尚不清楚。因此,我们在 47 例病例中进行了研究:i)使用识别细胞外结构域的抗体(Ext-Ab)和细胞内结构域特异性抗体(Int-Ab)通过免疫组织化学检测 EGFR 蛋白表达,ii)通过 RT-PCR 检测 EGFRv1、-v2、-v3、-v4 和 -vIII 的 mRNA 表达,以及 iii)通过荧光原位杂交检测 EGFR 扩增。研究了这些数据与组织学标准和患者预后的关系。Ext-Ab 的免疫染色比 Int-Ab 更强。EGFRv1、-v2、-v3 和 -v4 mRNA 表达高度相关。它们在所有肿瘤中均有表达,在胶质母细胞瘤中表达水平最高。EGFRv1 强水平和 vIII mRNA 的存在与 Int-Ab 染色更密切相关。EGFR 基因扩增仅涉及胶质母细胞瘤,与 EGFRvIII 的存在和 EGFRv2、-v3 和 -v4 转录本的高水平相关。不良预后与组织学(胶质母细胞瘤)、EGFR 扩增、Int-Ab 强烈标记和变体 mRNA 高水平相关。我们的结果表明,全长 EGFR 和突变型 EGFRvIII 并非弥漫性神经胶质瘤中唯一表达的 EGFR 异构体。这可以解释文献中报道的不一致的免疫组织化学结果,并可能具有治疗意义。