Quantitation of the Early Apoptotic Cells Using Flow Cytometry / 대한임상병리학회지

ABSTRACT

BACKGROUND: There is a strong evidence that administration of anti-tumor drugs triggers apoptotic death of target cells. Therefore, quantitation of the 0early apoptotic cells could provide a very useful information for clinicians planning anti-tumor strategies. Therefore, we measured the amount of early apoptotic cells using annexin-FITC/PI dual fuorescence method by which early changes of apoptotic cell membrane could be detected. We also measured DNA content of apoptotic cells by PI single stain and performed DNA fragment assay simultaneously. METHODS: HL-60 cell line were cultured under 100, 200 ng/mL adriamycin for 12, 24, 36, 48 hours. Quantitation of the early apoptotic cells was done using flow cytometry with annexin-FITC and Propidium iodide (PI) dual fluorescence stain. And DNA content of the HL-60 cells was measured using PI single stain after fixing the cells. DNA ladder assay was also performed by agarose gel electrophoresis. RESULTS: The early apoptotic cells were 40.5% at adriamycin 100 ng/mL, after 24 hours culture and the secondary necrotic cells were 94.7% at adriamycin 200 ng/mL after 48 hours culture. There was a good correlation between annexin-PI stain and DNA content analysis. We could find DNA fragmentation on agarose gel electrophoresis. CONCLUSION: Quantitation of the early apoptotic cells using flow cytometry with annexin-PI dual fluorochrome stain and the DNA content analysis with PI single stain could be a good parameter for anti-apoptotic strategies.