在高剂量甲氨蝶呤和亚叶酸素抢救治疗期间,在骨肉瘤患者的尿液中检测到碳酸酐酶II

Toyoji Sato, T. Kaneko, N. Ishikawa, Takako Toyama, S. Enomoto, T. Ōhashi, Yukio Shima, A. Kimura, H. Hatano, T. Uchiyama, Hiroyuki Segawa, Hiroto Kobayashi, T. Morita
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摘要

采用5-20%(线性梯度)聚丙烯酰胺平板凝胶电泳,研究了一名14岁骨肉瘤女孩在大剂量甲氨蝶呤(MTX)和亚叶酸素抢救治疗期间采集的尿蛋白。除白蛋白外,还观察到约30 kDa的明显蛋白带(P-30蛋白)。我们以前从未遇到过30kda的尿蛋白。因此,我们首先利用多肽质量指纹图谱和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对P-30蛋白进行了研究。我们得到的氨基酸序列为“YDPSLKPLSVSYDQATSLR”,与人类碳酸酐酶II (CAII)的氨基酸序列(40-58)相同。然后用抗人CAII抗体进行Western blotting证实该蛋白。这种酶广泛分布于人体内的红细胞、肾脏和胰腺中。我们认为这种CAII起源于肾脏,与大剂量甲氨蝶呤和亚叶酸素抢救治疗有关。
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Carbonic anhydrase II was detected in urine of a patient with osteosarcoma during high-dose methotrexate and leucovorin rescue therapy
Using a 5–20% (linear gradient) polyacrylamide slab gel electrophoresis, we studied the urinary protein collected from a 14-year-old girl with osteosarcoma during high-dose methotrexate (MTX) and leucovorin rescue therapy. A distinct protein band of approximately 30 kDa (P-30 protein) was observed in addition to albumin. We have never before encountered a 30 kDa urinary protein. Therefore, we first studied the P-30 protein by peptide mass fingerprinting and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). We obtained the amino acid sequence “YDPSLKPLSVSYDQATSLR”, which is the same as that of amino acids (40–58) of human carbonic anhydrase II (CAII). Then, we confirmed this protein by Western blotting using anti-human CAII antibody. This enzyme distributes widely in the human bodies, in erythrocytes, kidney, and pancreas. We considered that this CAII originated from the kidney in association with high-dose MTX and leucovorin rescue therapy.
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