Moore J P, Hancock D C, Littlewood T D, Evan G I
Ludwig Institute for Cancer Research, MRC Centre, Cambridge, U.K.
Oncogene Res. 1987;2(1):65-80.
The c-myc and N-myc nuclear oncoproteins are implicated in the genesis and maintenance of the transformed phenotype in several types of neoplastic disease, and the c-myc protein is involved in the progression of normal cells through the cell cycle. We have designed and developed sensitive and quantitative ELISAs for these proteins. Myc proteins are captured from cell lysates by an antibody directed against a peptide sequence substantially conserved in all known myc proteins; the captured proteins are recognised by a specific anti-c-myc or anti-N-myc monoclonal antibody conjugated to alkaline phosphatase; bound alkaline phosphatase is measured with an extremely sensitive cycling enzyme system that generates a coloured end-product. The c-myc assay is calibrated using bacterially expressed human c-myc protein. We have used this assay to estimate the number of c-myc molecules in a range of normal and transformed cells of human, murine, and feline origin; to monitor increases in c-myc expression when quiescent cells are stimulated with growth factors; and to follow the decrease in c-myc protein levels when HL60 promyelocytic leukaemia cells are induced to differentiate with dimethylsulphoxide or phorbol esters.
c-myc和N-myc核癌蛋白与多种肿瘤性疾病中转化表型的发生和维持有关，并且c-myc蛋白参与正常细胞通过细胞周期的进程。我们已经设计并开发了针对这些蛋白的灵敏且定量的酶联免疫吸附测定（ELISA）方法。Myc蛋白通过一种针对所有已知Myc蛋白中基本保守的肽序列的抗体从细胞裂解物中捕获；捕获的蛋白由与碱性磷酸酶偶联的特异性抗c-myc或抗N-myc单克隆抗体识别；结合的碱性磷酸酶用一种极其灵敏的循环酶系统进行检测，该系统产生一种有色终产物。c-myc检测使用细菌表达的人c-myc蛋白进行校准。我们已使用该检测方法来估计人、鼠和猫来源的一系列正常细胞和转化细胞中c-myc分子的数量；监测静止细胞受到生长因子刺激时c-myc表达的增加；以及追踪HL60早幼粒细胞白血病细胞用二甲亚砜或佛波酯诱导分化时c-myc蛋白水平的降低。
