Iles R K, Ind T E, Chard T
Williamson Laboratory for Molecular Oncology, Joint Academic Department of Reproductive Physiology, St Bartholomew's Hospital Medical College, West Smithfield, London, UK.
Br J Cancer. 1994 Feb;69(2):274-8. doi: 10.1038/bjc.1994.51.
Placental and placental-like alkaline phosphatase (PLAP) levels in the culture media of 87 cell lines of neoplastic and 'normal' origin were measured by a conventional immunosorbent enzymatic assay (IAEA) and by a new immunoradiometric assay (IRMA). The IRMA detected immunoreactive PLAP in 37 of 80 (46%) human epithelial and germ cell cultures, while the IAEA detected PLAP in only 25 (33%). Of the 52 non-germ cell tumour cultures, the IRMA detected expression in 24 (46%) and the IAEA in only 16 (31%). In 17 cases (21%) the IRMA recorded levels double that of the IAEA, while in five cultures (6%) the reverse was true. The IRMA was much more robust than the IAEA and had considerably lower inter- and intra-assay coefficients of variation (3.75-8.5% vs 5.2-46%). Detection of PLAP(-like) expression by IAEA is dependent on neoplastic expression of enzymatically functional molecules and quantification assumes constant enzyme kinetics. PLAP-like material has a higher catalytic rate constant than PLAP and thus will give higher values on a stoichiometric basis in an IAEA. The higher detection rate and levels of PLAP-like material in neoplastic cultures when measured by the IRMA clearly demonstrate ectopic expression of non-enzymatic PLAP and PLAP-like genes. The incidence of PLAP(-like) expression by non-germ cell and possible germ cell tumours has been underestimated and its utility as a tumour marker should be re-examined using assays which measure antigen mass rather than phosphatase activity.
采用传统免疫吸附酶法(IAEA)和新型免疫放射分析法(IRMA)检测了87株源于肿瘤和“正常”组织的细胞系培养基中的胎盘碱性磷酸酶和胎盘样碱性磷酸酶(PLAP)水平。在80株人上皮细胞和生殖细胞培养物中,IRMA检测到37株(46%)有免疫反应性PLAP,而IAEA仅检测到25株(33%)。在52株非生殖细胞肿瘤培养物中,IRMA检测到24株(46%)有表达,IAEA仅检测到16株(31%)。在17例(21%)中,IRMA记录的水平是IAEA的两倍,而在5株培养物(6%)中情况则相反。IRMA比IAEA更稳定,其批间和批内变异系数显著更低(分别为3.75 - 8.5%和5.2 - 46%)。IAEA对PLAP(样)表达的检测依赖于具有酶功能分子的肿瘤表达,并且定量假设酶动力学恒定。PLAP样物质的催化速率常数高于PLAP,因此在IAEA中基于化学计量学它会给出更高的值。用IRMA测量时,肿瘤培养物中PLAP样物质的检测率和水平更高,这清楚地证明了非酶性PLAP和PLAP样基因的异位表达。非生殖细胞和可能的生殖细胞肿瘤中PLAP(样)表达的发生率被低估了,应使用测量抗原量而非磷酸酶活性的检测方法重新审视其作为肿瘤标志物的效用。