Akimoto J, Ikeda K, Tomonaga M, Saito F, Miwa T, Awaya A, Hashimoto Y, Fukui H
Department of Neuropathology, Faculty of Medicine, University of Tokyo.
No To Shinkei. 1991 Jan;43(1):25-9.
It is well known that some fetal antigens are expressed in malignant tumor cells. Likewise, brain tumors, especially histologically malignant cases, may have any antigenic relationships with fetal brain. So, we investigated the relationship by immunohistochemical technique, utilizing a polyclonal antibody to mouse fetal stage-specific polypeptide "GP68". We prepared GP68 from homogenate of head part of embryos at the 14th day of gestation mice by RCA-1 agarose column chromatography. And immunized it to Japanese white rabbits and the titer was measured by enzyme-linked immunosorbent assay. We analyzed operatively resected brain tumors and autopsy brain tissues. Frozen tissues were fixed in cold acetone and immunostained with anti-GP68 serum according to biotin-streptavidin peroxidase method. Remained tissues were homogenized in Laemmli's sample buffer and electrophoresed. The proteins were transferred to nitrocellulose membrane and immunostained with anti-GP68. Normal brain tissues were not positively stained, except for capillary endothelium which showed a weak staining. On the other hand, brain tumors of neuroectodermal origin were positively stained in varying degrees, and other tumors were negative. It is especially noteworthy that, in astrocytoma cases, there exists a definite correlation between the intensity of stain and the degree of histological malignancy. Immunoblot studies demonstrated a very weak band at 68 KD in normal brain and meningioma. In contrast, very strong band at the same position was seen in malignant astrocytomas. These results suggested that in brain tumors, especially those of neuroectodermal origin, GP68 antigen is expressed and the degree of expression is related to their histological malignancy. So this fetal antigen may be useful for evaluation of biological malignancy of gliomas.(ABSTRACT TRUNCATED AT 250 WORDS)
众所周知，一些胎儿抗原在恶性肿瘤细胞中表达。同样，脑肿瘤，尤其是组织学上的恶性病例，可能与胎儿脑存在任何抗原关系。因此，我们利用针对小鼠胎儿阶段特异性多肽“GP68”的多克隆抗体，通过免疫组织化学技术研究了这种关系。我们通过RCA-1琼脂糖柱色谱法从妊娠第14天小鼠胚胎头部匀浆中制备了GP68。并将其免疫日本白兔，通过酶联免疫吸附测定法测量效价。我们分析了手术切除的脑肿瘤和尸检脑组织。将冷冻组织用冷丙酮固定，并根据生物素-链霉亲和素过氧化物酶法用抗GP68血清进行免疫染色。剩余组织在Laemmli样品缓冲液中匀浆并进行电泳。蛋白质转移到硝酸纤维素膜上并用抗GP68进行免疫染色。正常脑组织除毛细血管内皮有弱阳性染色外，无阳性染色。另一方面，神经外胚层起源的脑肿瘤有不同程度的阳性染色，其他肿瘤为阴性。特别值得注意的是，在星形细胞瘤病例中，染色强度与组织学恶性程度之间存在明确的相关性。免疫印迹研究显示正常脑和脑膜瘤在68KD处有一条非常弱的条带。相比之下，在恶性星形细胞瘤中在相同位置可见非常强的条带。这些结果表明，在脑肿瘤中，尤其是神经外胚层起源的肿瘤，GP68抗原表达，且表达程度与其组织学恶性程度相关。因此，这种胎儿抗原可能有助于评估胶质瘤的生物学恶性程度。（摘要截短至250字）
