DNA and nuclear protein measurement in columnar epithelial cells of human endometrium

Abstract

Propidium iodide DNA flow cytometry, Feulgen-DNA, and nuclear light green protein scanning cytometry were performed in columnar epithelial cells of normal, nonmalignant human endometrium and endometrial adenocarcinomas. Columnar cells were identified by immunohistochemical staining for cytokeratin 18, an intermediate filament protein specifically present in columnar cell epithelium. DNA measurements derived from flow and scanning cytometry showed comparable results. The DNA content of the GO/Gl fraction of the adenocarcinomas had a considerable overlap with that of normal endometrium, with that of the carcinomas shifted toward higher values. For the carcinomas, no correlation was found with the histological grade, with the exception of the adenosquamous carcinomas. Most of the clinical stage I tumors showed a DNA content in the normal diploid region. Three of the four carcinomas of clinical stage I1 and higher had an increased DNA content.For the carcinomas, the percentage of cells in the proliferative fraction, as determined In a preceding paper (111, we compared DNA and nuclear protein values in normal endometrium and endometrial adenocarcinomas to study the changes herein during neoplastic transformation. The results showed that the carcinoma cells had the same or a n increased DNA content compared with normal endometrium, a large variation in the percentage of cells in the proliferative fraction, and a comparable amount of nuclear protein. These measurements were performed in isolated nuclei obtained from homogenates of normal endometrium and endometrial adenocarcinomas. Normal endofrom scanning cytometric derived DNA histograms, was comparable to that of normal endometrium, or higher. No correlation was found with the histological grade. Tumors of clinical stage I1 and higher had intermediate values compared to carcinomas of lower stages.The nuclear protein/DNA ratio of malignant endometrium completely overlapped that of normal endometrium. Within the tumor poy;,ulation, no correlation was found with the hisitological grade, with the exception of the adenosquamous carcinomas, and clinical stage.Based on the aforementioned parameters, no discrimination could be obtained between normal and malignant endometrium. However, when the DNA content of the GO/G1 fraction was combined with the coefficient of variation of the nuclear protein/UNA ratio, a clear discrimination could be obtained with only two false-positive cases.Key terms: Adenocarcinomas, DNA cytometry, nuclear protein cytometry, cytokeratin metrium and endometrial adenocarcinomas are composed of several tissue types, however. Normal endometrium contains a varying number of glands, dependent. on the period of the menstrual cycle, which are lined up by columnar epithelial cells. The glands are Address reprint requests to Peter S. Oud,

Comprehensive Analysis of Yeast Surface Displayed cDNA Library Selection Outputs by Exon Microarray to Identify Novel Protein-Ligand Interactions

“…The amount of PtdIns (4,5) in detergent-washed rat liver nuclei was measured as ϳ30 pmol/mg of protein (40). Using an estimate of 72 mg/ml (calculated using a value of 3.1 pg for the mass of the haploid nuclear DNA content of the rat genome (49) and a 2:1 nuclear protein:DNA mass ratio (50,51) for the total nuclear protein concentration gives a rough estimate of 4 M for the nuclear PtdIns(4,5) concentration, indicating that interactions in the affinity range we observed could be biologically significant. Our experiments demonstrated that phosphatidylinositides and double-stranded DNA oligonucleotides derived from homeobox domain target sequences compete for binding to homeobox domain-containing protein fragments.…”