Schut I C, Waterfall P M, Ross M, O'Sullivan C, Miller W R, Habib F K, Bayne C W
Breast Cancer Research Group, Department of Oncology, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XU, Scotland, UK.
BJU Int. 2003 Feb;91(3):278-83. doi: 10.1046/j.1464-410x.2003.03062.x.
To detect the expression and transcription pattern of MUC1 in benign and malignant disease, and in two widely studied cell lines, and to investigate the glycosylation of MUC-1 in bone metastasis of prostate cancer, as mucins have been implicated in the progression and behaviour of several cancers.
RNA extracted from cell lines (DU145 and PC3), five samples of BPH and five samples of prostate cancer was reverse transcribed before amplification of MUC1-specific sequences by polymerase chain reaction. Paraffin-embedded sections were stained for glycosylated MUC1 and MUC1 core epitopes by HMFG1 and B27.29 antibodies, respectively. Steroid-treated cell lines were analysed by fluorescence-activated cell sorting, using the same antibodies.
MUC1, in an under-glycosylated form, was widely expressed in the prostate and in metastatic lesions. MUC1/Z and MUC1/Y RNA were differentially expressed in BPH and prostate cancer, with no detectable expression of splice variant mRNA. This is in contrast to prostate cancer cell line cells (PC3 and DU145), which express splice variant mRNA.
BPH, prostate cancer and metastatic prostate cancer all express high levels of under-glycosylated MUC1. This may explain the inability of previous studies to detect MUC1 in prostate tissue, as the antibody used was specific for a carbohydrate epitope which is not expressed on the under-glycosylated MUC1.
检测黏蛋白1(MUC1)在良性和恶性疾病以及两种广泛研究的细胞系中的表达和转录模式,并研究前列腺癌骨转移中MUC-1的糖基化情况,因为黏蛋白与多种癌症的进展和行为有关。
从细胞系(DU145和PC3)、5份良性前列腺增生(BPH)样本和5份前列腺癌样本中提取RNA,在通过聚合酶链反应扩增MUC1特异性序列之前进行逆转录。石蜡包埋切片分别用HMFG1和B27.29抗体对糖基化MUC1和MUC1核心表位进行染色。使用相同抗体,通过荧光激活细胞分选对经类固醇处理的细胞系进行分析。
低聚糖基化形式的MUC1在前列腺和转移灶中广泛表达。MUC1/Z和MUC1/Y RNA在BPH和前列腺癌中差异表达,未检测到剪接变体mRNA的表达。这与表达剪接变体mRNA的前列腺癌细胞系(PC3和DU145)形成对比。
BPH、前列腺癌和转移性前列腺癌均表达高水平的低聚糖基化MUC1。这可能解释了先前研究在前列腺组织中未能检测到MUC1的原因,因为所用抗体对低聚糖基化MUC1上未表达的碳水化合物表位具有特异性。