Chan P K, Qi Y, Amley J, Koller C A
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA.
Cancer Lett. 1996 Feb 27;100(1-2):191-7. doi: 10.1016/0304-3835(95)04100-1.
We have previously detected by immunofluorescent assay that the cellular localization of nucleophosmin/B23 (NPM) shifts from the nucleolus to the nucleoplasm (NPM-translocation) after exposure of cells to multiple agents. In order to improve the quantification of the NPM-translocation, we have developed a digital imaging technique. Human Lo leukemia cells, MCF-7 breast carcinoma cells, and fresh human leukemia cells were exposed to anthracyclines or actinomycin D for 4 h. The degree of NPM-translocation was determined and presented as the localization index (LI). Control cells had a LI of about 10, which indicates that the majority of NPM was localized in nucleoli. The LI for drug-treated cells decreased in a dosage- and time-dependent manner. The effect of two classes of anthracycline (daunomycin and aclacinomycin A) and different types of intercalators (daunomycin and actinomycin D) had additive effects on induction of NPM-translocation. The imaging procedure was easily applied to fresh leukemia cells, thus providing useful information regarding drug effects on cancer cells.
我们之前通过免疫荧光测定法检测到，细胞在接触多种试剂后，核磷蛋白/B23（NPM）的细胞定位从核仁转移至核质（NPM易位）。为了改进对NPM易位的定量分析，我们开发了一种数字成像技术。将人Lo白血病细胞、MCF-7乳腺癌细胞和新鲜人白血病细胞暴露于蒽环类药物或放线菌素D中4小时。测定NPM易位程度并将其表示为定位指数（LI）。对照细胞的LI约为10，这表明大多数NPM定位于核仁。药物处理细胞的LI呈剂量和时间依赖性降低。两类蒽环类药物（柔红霉素和阿克拉霉素A）和不同类型嵌入剂（柔红霉素和放线菌素D）对诱导NPM易位具有相加作用。该成像程序易于应用于新鲜白血病细胞，从而提供有关药物对癌细胞作用的有用信息。
