胸苷激酶1抗体对细胞增殖的影响

Naining Wang, Qimin He, S. Skog, S. Eriksson, B. Tribukait
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引用次数: 38

摘要

胞质胸苷激酶1 (TK1)是参与DNA复制的酶之一。生化研究表明,TK1在细胞周期G1后期被激活,其活性与细胞增殖有关。我们开发了一种多克隆抗TK1抗体,针对从人TK1 C端合成的肽。利用这种抗体,我们证明了TK1在细胞细胞质中的排他性位置。通过同时对TK1和溴脱氧尿嘧啶进行荧光染色,利用洗脱的细胞,定量TK1的量与细胞DNA含量的关系,研究细胞周期依赖性TK1的表达。TK1在S+G2时期强烈表达,在G1晚期升高,在有丝分裂期间降低。从G1晚期到G2, TK1的数量增加了3倍。TK1阳性细胞在各种正常和恶性组织中均有增殖活性。新的抗TK1抗体在档案标本中起作用,是细胞增殖的特异性标志物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Investigation on Cell Proliferation with a New Antibody against Thymidine Kinase 1
The cytosolic thymidine kinase 1 (TK1) is one of the enzymes involved in DNA replication. Based on biochemical studies, TK1 is activated at late G1 of cell cycle, and its activity correlates with the cell proliferation. We have developed a polyclonal anti‐TK1 antibody against a synthetic peptide from the C‐terminus of human TK1. Using this antibody, here we demonstrate the exclusive location of TK1 in the cytoplasm of cells. Cell cycle dependent TK1 expression was studied by simultaneous fluorescence staining for TK1 and bromodeoxyuridine, by using elutriated cells, and by quantitation of the amount TK1 in relation to the cellular DNA content. TK1, which was strongly expressed in the cells in S+G2 period, raised at late G1 and decreased during mitosis. The amount of TK1 increased three folds from late G1 to G2. TK1 positive cells were demonstrated in areas of proliferation activity of various normal and malignant tissues. The new anti‐TK1 antibody works in archival specimens and is a specific marker of cell proliferation.
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