Diagnostic method for differentiation of hepatocellular carcinoma : Determination of immature TGFα in blood and urine.

ProTGFα被加工成TGFα的成熟形式。本研究合成了几种氨基酸序列仅存在于proTGFα序列中而不存在于成熟TGFα序列中的多肽。用其中一种肽免疫新西兰白色兔。我们用抗血清中的免疫球蛋白G组分和抗TGFα的多克隆抗体建立了夹心ELISA。从正常成人或肝细胞癌患者收集的尿液中加入终浓度为70%的硫酸铵并离心。将酒石酸盐级分用作测定样品。新开发的ELISA没有检测到TGFα的成熟形式。然而，它检测到TGFα的未成熟形式，其分子量大于成熟形式。在肝细胞癌患者尿中检测到未成熟型TGFα，而在正常成人尿中未检测到。根据这些结果，肝细胞癌患者中未成熟形式的TGFα可能升高。检测未成熟型TGFα对肝癌的诊断有一定意义。我们现在正在分析肝细胞癌患者血液中未成熟形式的TGFα的含量。

ProTGFα is processed to become a mature form of TGFα. In this study, we synthesized several kinds of peptides, amino acid sequences of which exist only in the sequence of proTGFα but not in that of the mature form of TGFα. New Zealand white rabbits were immunized with one of the peptides. We developed a sandwich ELISA using immunoglobulin G fraction of the antiserum and polyclonal antibodies against TGFα. Urine collected from normal adults or patients with hepatocellular carcinoma was added with ammonium sulfate at the final concentration of 70 percent and centrifuged. Precipitate fraction was used as an assay sample. The newly developed ELISA did not detect the mature form of TGFα. However, it detected an immature form of TGFα, the molecular weight of which was larger than that of the mature form. The immature form of TGFα was detected in urine from patients with hepatocellular carcinoma but not in that from normal adults. From these results, it is possible that the immature form of TGFα is elevated in patients with hepatocellular carcinoma. Determination of the immature form of TGFα can be useful for diagnosis of hepatocellular carcinoma. We are now analyzing the content of the immature form of TGFα in blood from patients with hepatocellular carcinoma.