Wani G, Wani A A, D'Ambrosio S M
Department of Radiology, College of Medicine, Ohio State University, Columbus 43210.
Carcinogenesis. 1992 Mar;13(3):463-8. doi: 10.1093/carcin/13.3.463.
The cell-type-specific expression of human O6-methylguanine-DNA methyltransferase (O6-MT) was determined in paraffin-embedded sections of human kidney. A 39 base oligomer complementary to O6-MT cDNA was labeled with digoxigenin and visually detected in situ using an alkaline phosphatase-conjugated anti-digoxigenin antibody. This allowed direct determination of O6-MT-specific mRNA levels, while simultaneously identifying the structures and cell types in the kidney section. Expression of O6-MT was high in distal tubular and glomerular epithelial cells and low in the cells of the Bowman's capsule, collecting and proximal tubular cells. Hybridization of the oligomer was specific to RNA, since RNase and not DNase eliminated the signal. Expression was uniform in all the cell types, except the glomerular cells exhibited varying levels of high intensity. Cell-specific expression was constant between tissue sections from the same and different kidney tissues. These data may help explain the differential response of various cell types to alkylating agents.
在人肾脏石蜡包埋切片中测定了人O6-甲基鸟嘌呤-DNA甲基转移酶（O6-MT）的细胞类型特异性表达。用洋地黄毒苷标记与O6-MT cDNA互补的39个碱基的寡聚物，并使用碱性磷酸酶偶联的抗洋地黄毒苷抗体进行原位可视化检测。这使得能够直接测定O6-MT特异性mRNA水平，同时识别肾脏切片中的结构和细胞类型。O6-MT在远端肾小管和肾小球上皮细胞中表达较高，而在鲍曼囊、集合管和近端肾小管细胞中表达较低。寡聚物的杂交对RNA具有特异性，因为核糖核酸酶而非脱氧核糖核酸酶消除了信号。除了肾小球细胞呈现出不同程度的高强度外，所有细胞类型中的表达都是均匀的。来自相同和不同肾脏组织的组织切片之间，细胞特异性表达是恒定的。这些数据可能有助于解释各种细胞类型对烷化剂的不同反应。
