Induction of differentiation in the human leukemia cell line SPI‐802: Morphological, immunological, and isoenzymatic changes

Abstract

The phorbolester 12-0-tetradecanoylphorbol 13-acetate (TPA) was used for the induction of differentiation in cells of the human leukemia cell line SPI-802. The cellular morphology, surface marker antigen expression, and isoenzyme profiles of four enzymes (carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase) served as parameters for monitoring the induced phenotypical changes. TPA led to distinct alterations of the morphology and significantly affected the growth rate with cessation of cell proliferation. No major increase in the number of nitro blue tetrazolium-positive cells or aggregation of cells, phagocytosis of latex beads, adherence to plastic surface, or development of pseudopodia were observed. As TPA-treated SPI-802 cells remained negative for these markers of the monocyte-macrophage complex, it can be concluded that the cells did not differentiate into monocytes and macrophages. The immunological marker profile based on testing of 55 monoclonal antibodies, terminal deoxynucleotidyl transferase and two erythrocyte rosette tests indicated a differentiation of SPI-802 cells along the granulocytic cell lineage. This was confirmed by isoenzyme analysis, especially that of carboxylic esterase. An isoenzyme specific for monocytes and macrophages was not detected. In earlier studies it was found that SPI-802 cells produce hemoglobin upon exposure to TPA or hemin. This latter observation and the present results suggested a comparison with the two erythroleukemia cell lines K-562 and HEL. SPI-802 cells appear to have the potential to differentiate along several cell lineages.

Evidence for activated Lck protein tyrosine kinase as the driver of proliferation in acute myeloid leukemia cell, CTV-1

“…In this study, we use small molecule inhibitors against various oncogenic protein tyrosine kinases to probe for the key drivers of proliferation in two M5 AML cells, CTV-1 [23][24] and THP-1 [25][26]. The inhibition profiling suggests that Lck protein tyrosine kinase is the key oncogenic driver of CTV-1 cell proliferation, which is confirmed by molecular and mutagenic analyses.…”

“…To identify the drivers of proliferation in the two M5 AML cell lines, CTV-1 and THP-1, we determined their sensitivity to a panel of PTK inhibitors. CTV-1 is a human monocytic leukemia cell line established from a 40 yr-old male patient with relapsed acute monoblastic leukemia [23][24]. THP-1 is a monocytic cell line established from a 1-yr old male patient of acute monocytic leukemia [25].…”

“…CTV-1 cell line was established as a human monocytoid leukemia cell line from a patient with relapsed monoblastic leukemia [23]. The cell can be induced to differentiate by the phorbolester 12-0-tetradecanoylphorbol 13-acetate treatment [24]. Surface marker analysis with 45 monoclonal antibodies and several polyclonal antisera and isoenzyme analysis of carboxylic esterase, acid phosphatase, and hexosaminidase and lactate dehydrogenase demonstrated CTV-1 to be of monocytoid origin arrested at a very early stage of differentiation [24].…”

Activating mutations in NOTCH1 in acute myeloid leukemia and lineage switch leukemias

“…This analysis demonstrated the presence of truncating mutations in the PEST domain of NOTCH1 in three cell lines. CTV-1 cells, a minimally differentiated myeloid line derived from an AML M5 patient at relapse, 13,14 showed a point mutation (7580C/A), generating a premature stop codon at position 2527 in NOTCH1 (Figure 1c). In addition, two myelo-monocytic (AML M4) sister cell lines, ML1 and ML2, harbor a frameshift mutation as a result of a dinucleotide deletion at positions 7550-7551 (Figure 1d).…”