Molecular and immunological approach to hematological disease: detection and analysis of intracellular modified nucleosides by flowcytometory

I Honda, MD1, K Itou, PhD2, M Mizugaki, PhD2, T Sasakati MD2

1National Iwate Hospital, 2Tohoku University School of Medicine

AIM: Modified nucleosides are components of ribosomal RNA(rRNA),transfer RNA (tRNA) and messenger RNA(mRNA). 1-methyladenosine and pseudouridine are members of those modified nucleosides. The urinary concentration of 1-methyladenosine and pseudouridine of cancer patients are higher than those of healthy controls, and those compounds are reduced after effective chemotherapy. Thus those compunds might be expected to use as tumor markers. ?METHOD?In this study cellular origin of 1-methyladenosine and pseudouridine were analysed about two tumor cell lines (HUT-102,THP-1),peripheral lymphocytes (PBL) from healthy adult and PBL under the phytohemagglutinin stimulation, by flowcytometric analysis and immunofluorescent staining of cellular RNA using monoclonal antibodies specific for 1-methyladenosine (AMA) and pseudouridine (APU). ?RESULTS? Both 1-methyladenosine and pseudouridine were detected in more than 90% of tumor cells above the thresholds of flowcytemetric detection(Spectrum III,Ortho).The PBL under the PHA stimulation also tended to take the same way of the tumor cell lines, whereas few of the PBL contained 1-methyladenosine above the thresholds. ?CONCLUSIONS?According to the DNA analysis of those cell lines, high contents of the modified nucleosides in the cell might follow DNA synthesis, this leads to one reason for high levels of the urinary excretion of the modified nucleosi-des in cancer patients.

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Paper presented at the International Symposium on Predictive Oncology and Intervention Strategies; Paris, France; February 9 - 12, 2002; in the section on Molecular Pathology.