Karasaki S, Okigaki T
Cancer Res. 1976 Dec;36(12):4491-9.
A cell surface-located nucleoside triphosphatase activity can be assayed in liver epithelial cultures in situ with the incubation of intact cells in a medium containing [gamma-32P]adenosine triphosphate and correlated with the tumorigenicity of these cells in neonatal Wistar rats. The ectoenzyme activity of normal diploid cell lines is minimal, whereas a considerably high activity has been found in all tumorigenic cell lines tested. The optimum condition for the adenosinetriphosphatase activity is physiological with regard to osmolarity, ionic composition, pH, and substrate concentration in the medium. The enzyme is significantly stimulated by Ca2+, and its activation is controlled by Mg2+. Histochemical examinations indicate that glutaraldehyde-fixed cells of tumorigenic lines have Ca2+-stimulated adenosinetriphosphatase activity on the external surface. The isotopic assay of adenosine triphosphate hydrolysis by intact cells may provide a rapid method for screening oncogenesis in vitro of liver epithelial cells.
细胞表面定位的核苷三磷酸酶活性可在肝上皮细胞原位培养物中进行测定,方法是将完整细胞在含有[γ-32P]三磷酸腺苷的培养基中孵育,并与这些细胞在新生Wistar大鼠中的致瘤性相关联。正常二倍体细胞系的外切酶活性最低,而在所有测试的致瘤细胞系中均发现了相当高的活性。就培养基的渗透压、离子组成、pH值和底物浓度而言,三磷酸腺苷酶活性的最佳条件是生理性的。该酶受到Ca2+的显著刺激,其激活由Mg2+控制。组织化学检查表明,致瘤细胞系经戊二醛固定的细胞在外表面具有Ca2+刺激的三磷酸腺苷酶活性。完整细胞对三磷酸腺苷水解的同位素测定可能为筛选肝上皮细胞体外肿瘤发生提供一种快速方法。