Transferrin receptor expression during exponential and plateau phase growth of human tumour cells in culture.
Transferrin receptor expression by the human tumour cell lines CCRF-CEM leukaemia and PMC-22B melanoma was studied, measuring the specific binding of fluorescein isothiocyanate (FITC)-labelled transferrin using a fluorescence-activated cell sorter. By measuring the fluorescence of cells stained at subsaturating concentrations of conjugate it was possible to calculate the average numbers of receptors per cell and the binding affinity by Scatchard analysis. These values (1.9 X 10(5) binding sites/cell, KA 1.2 X 10(9) M-1 for CCRF-CEM during exponential growth and 6.9 X 10(4) binding sites/cell, KA 1.4 X 10(-9) M-1 for PMC-22B) are in close agreement with previously published data obtained using radiolabelled transferrin. The present method, however, allowed the transferrin receptor expression of individual cells within a population to be measured and thus it has been possible to test the hypothesis that transferrin receptor is a marker for cycling cells. Frequency-distribution histograms of transferrin receptor showed a wide range of values for both cell lines during exponential growth. When the extreme ranges were sorted and the cells examined for cellular DNA content it was found that those with the highest transferrin receptor expression were enriched with cells in S, G2, and M phases of the cell cycle, whereas those with low transferrin receptor expression were mainly in G1. However, two-parameter-correlated dot plots of transferrin receptor expression versus DNA content showed there was considerable overlap between the ranges of receptor expression for the different cell cycle compartments. Using a stathmokinetic method we have measured the proportion of quiescent cells in fed plateau phase cultures. Transferrin receptor expression was downgraded under these growth conditions but, contrary to expectation, the decline affected the population uniformly, without the emergence of a distinct, transferrin receptor-negative subpopulation corresponding to the increasing proportion of quiescent cells. Thus, although transferrin receptor expression bears some relation to cell cycle phase and reflects the proliferative activity of populations of cells, it is incapable of identifying individual cells which are out of cycle.
D001059:Apoproteins; D002453:Cell Cycle; D002455:Cell Division; D002478:Cells, Cultured; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006801:Humans; D009369:Neoplasms / Q000032:analysis* / Q000473:pathology; D011956:Receptors, Cell Surface / Q000032:analysis*; D011990:Receptors, Transferrin; D013861:Thiocyanates; D014168:Transferrin / Q000032:analysis
D001059:Apoproteins; D005452:Fluoresceins; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D013861:Thiocyanates; D014168:Transferrin; C028284:apotransferrin; D016650:Fluorescein-5-isothiocyanate