Flow cytometric determination of tumor ploidy and S-phase fraction following collagenase dissociation and thymidine labeling was performed on 75 consecutive breast cancers. Estrogen and progesterone receptor levels and routine histologic examination also were obtained on each tumor. Cell viability following collagenase dissociation varied from 13 to 95% with a mean of 71%. Thirty-six tumors were diploid, four tetraploid, and four hypertetraploid, and the remainder had DNA indices between 1.1 and 1.9. There was no significant correlation between tumor ploidy and tumor size or estrogen receptor positivity or negativity. The percentage of cells in S-phase varied from 1.2 to 20.0% with a mean of 6.0% utilizing a rectilinear model for histogram analysis that integrated a 10-contiguous channel sample containing the lowest number of cells in S-phase (S-pFL). The mean S-pFL of diploid carcinomas (3.43%) was significantly lower than that of hyperdiploid carcinomas (8.38%). There was good correlation between S-phase fraction determined by thymidine-labeling index (TLI) and S-pFL (r = 0.772, p = 0.0001). S-pFL predicted whether a tumor would be above or below median TLI with an accuracy of 90.5%. Estrogen receptor-negative cancers tended to have higher TLIs and S-pFLs than estrogen receptor-positive cancers; however, there was no correlation between progesterone receptor positivity or negativity and TLI and S-pFL.
Download full-text PDF |
Source |
---|
Front Immunol
January 2025
Laboratory of Fish Protistology, Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice, Czechia.
Methods Mol Biol
November 2024
Department of Biological Informatics and Experimental Therapeutics, Graduate School of Medicine, Akita University 1-1-1, Hondo, Akita, Japan.
This protocol describes the detection of phosphorylated proteins within cells and the identification of their intracellular localization, with a particular focus on mitotic cells. While the detection of phosphorylated proteins can be achieved using radioactive labeling with Pi, this method presents experimental challenges due to the requirement for radioisotopes. Alternatively, detection using phosphorylation-specific antibodies is a potent method; however, it necessitates the identification of phosphorylation sites and further requires the generation of antibodies targeting these sites, making it effective only for thoroughly analyzed phosphorylations.
View Article and Find Full Text PDFRadiat Prot Dosimetry
November 2024
Laboratory of Radiological Disasters and Medical Science, International Research Institute of Disaster Science, Tohoku University, 519-1176 Aramaki-aza-Aoba, Aoba-ku, Sendai 980-0845, Japan.
This study investigated the induction of DNA double-strand breaks (DSBs) in the hTERT-immortalized normal human diploid epithelial cells (RPE1-hTERT) continuously exposed to 6000 Bq/ml of tritiated water (HTO) and organically bound tritium (OBT). The relationship of the DSBs induction with the intracellular amount as well as the localization of tritium was also examined. Tritium-labeled thymidine (3H-Thy) and palmitic acid (3H-PA) were used as OBT.
View Article and Find Full Text PDFSTAR Protoc
December 2024
Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA. Electronic address:
The cardiac regeneration and development fields lack low-barrier-to-entry techniques that distinguish cardiomyocyte division from alternative outcomes in vivo. Here, we present a protocol in rodents to determine if cardiomyocyte cell division has occurred. We describe thymidine analog administration, Langendorff procedure, immunofluorescent labeling, microscopy, and analysis of fluorescent images to assess ploidy, thereby allowing an investigator to retrospectively claim cell division.
View Article and Find Full Text PDFJ Org Chem
December 2024
Center for Advanced Technology, Adam Mickiewicz University, Poznań 61-614, Poland.
DNA photo-crosslinking reactions occur widely in biological systems and are often used as valuable tools in molecular biology. In this article, we demonstrate the application of an oligonucleotide 5-fluoro-2'-O-methyl-4-thiouridine ()-containing probe for the fluorescent detection of specific DNA sequences. The design of the probe was predicated on studies of agents that could adversely affect its efficiency.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!