Lloyd R V, Jin L, Fields K
Department of Pathology, University of Michigan, Ann Arbor 48109-0054.
Am J Surg Pathol. 1990 Jan;14(1):35-43. doi: 10.1097/00000478-199001000-00004.
We analyzed the distribution of chromogranins A and B in normal and neoplastic endocrine tissues with secretory granules using 35S-labeled and biotin-labeled oligonucleotide probes by in situ hybridization (ISH). Both radioactive and nonradioactive probes detected messenger RNAs (mRNAs) in frozen and paraffin tissue sections. Endocrine tissues with variable immunoreactivities for chromogranin A protein, such as small-cell lung carcinomas, neuroblastomas, insulinomas, and parathyroid adenomas, expressed the mRNA for chromogranins A and B in most cells. Some technical problems with the biotinylated probes included nonspecific nuclear staining and endogenous alkaline phosphatase, which was not completely abolished by levamisole pretreatment. A differential distribution of chromogranins A and B was seen in pituitary prolactinomas, which expressed abundant chromogranin B but not chromogranin A mRNAs, and in parathyroid adenomas, which expressed abundant chromogranin A but only small amounts of chromogranin B mRNAs. These results indicate that ISH can be used to detect chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes and that the mRNAs for chromogranin A and B are demonstrable in some tumors even when the chromogranin proteins cannot be detected by immunohistochemistry.
我们采用原位杂交（ISH）技术，使用35S标记和生物素标记的寡核苷酸探针，分析了正常和肿瘤性内分泌组织中具有分泌颗粒的嗜铬粒蛋白A和B的分布情况。放射性和非放射性探针均可在冰冻和石蜡组织切片中检测到信使核糖核酸（mRNA）。对嗜铬粒蛋白A蛋白具有不同免疫反应性的内分泌组织，如小细胞肺癌、神经母细胞瘤、胰岛素瘤和甲状旁腺腺瘤，在大多数细胞中均表达嗜铬粒蛋白A和B的mRNA。生物素化探针存在一些技术问题，包括非特异性核染色和内源性碱性磷酸酶，左旋咪唑预处理不能完全消除这些问题。在垂体催乳素瘤中观察到嗜铬粒蛋白A和B的差异分布，垂体催乳素瘤表达丰富的嗜铬粒蛋白B但不表达嗜铬粒蛋白A的mRNA；在甲状旁腺腺瘤中也观察到差异分布，甲状旁腺腺瘤表达丰富的嗜铬粒蛋白A但仅表达少量的嗜铬粒蛋白B的mRNA。这些结果表明，ISH可用于用放射性和生物素化寡核苷酸探针检测内分泌组织中的嗜铬粒蛋白A和B，并且即使免疫组织化学检测不到嗜铬粒蛋白，在某些肿瘤中仍可检测到嗜铬粒蛋白A和B的mRNA。
