A Study of Differentiation Using HL60 Cells as a Model

Mitchell, Lynn Shona (1987) A Study of Differentiation Using HL60 Cells as a Model. PhD thesis, University of Glasgow.

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HL60 is a human promyelocytic, leukaemic cell line. The cells are bipotent in culture; treatment with retinoic acid (RA) or dimethyl sulphoxide (DMSO) induces HL60 cells to differentiate to granulocytes whereas treatment with 12-O-tetradecanoyl 13-phorbol acetate (TPA) induces these cells to differentiate to monocytes / macrophages. Differential screening of a cDNA library representing poly(A)+ RNA from RA induced HL60 cells led to the selection of three different probes for sequences which change in abundance following differentiation of HL60 cells (F6, F10, C6). F6 RNA is highly abundant in uninduced cells but not in induced HL60 cells. Sequencing of F6 cDNA revealed that it consisted of an Alu repetitive element. Both F10 and C6 RNAs increase in abundance during differentiation of HL60 cells to granulocytes. In situ hybridisation indicated that F10 RNA represents a differentiation stage-specific transcript of the myeloid lineage. A probe for C6 RNA detected two transcripts. Analysis of the distribution of these RNAs between nuclear and total RNA isolated from RA induced HL60 cells suggested that the larger transcript is an unprocessed precursor of the smaller C6 RNA. The timing of changes in expression of these three genes, together with c-myc and lysozyme, has been determined during the induced differentiation of HL60 cells. This has been compared to the timing of the commitment event, as determined by examination of parameters which assess the cell's potential to self-renew and their differentiation status. It appears that c-myc expression bears a direct relationship to proliferation and differentiation of TPA induced HL60 cells. However, no such relationship was observed during DMSO and RA induction. A direct relationship was also observed between C6 and F10 expression and differentiation of HL60 cells to granulocytes.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Keywords: Cellular biology, Oncology
Date of Award: 1987
Depositing User: Enlighten Team
Unique ID: glathesis:1987-77620
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 14 Jan 2020 11:53
Last Modified: 14 Jan 2020 11:53
URI: https://theses.gla.ac.uk/id/eprint/77620

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