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Induction of human malignant T‐lymphoblastic cell lines MOLT‐3 and jurkat by 12‐O‐tetradecanoylphorbol‐13‐acetate: Biochemical, physical, and morphological characterization
Author(s) -
Nagasawa Kohei,
Howatson Allan,
Mak Tak W.
Publication year - 1981
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041090120
Subject(s) - 12 o tetradecanoylphorbol 13 acetate , jurkat cells , antipain , cell culture , microbiology and biotechnology , lymphoblast , terminal deoxynucleotidyl transferase , tetradecanoylphorbol acetate , acetylation , t cell , biology , chemistry , medicine , endocrinology , biochemistry , immunology , apoptosis , enzyme , protein kinase c , phorbol ester , tunel assay , genetics , immune system , leupeptin , protease , gene
The process of induction of human malignant T‐lymphoblastic cell line MOLT‐3 by the phorbol ester 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) was examined. It was found that the induction process by TPA, which included increase in cells with receptors to sheep red blood cells (E‐rosette positive‐E + ) and decrease in the levels of the marker enzyme terminal deoxynucleotidyl transferase (TdT) was not affected by the presence of DNA synthesis inhibitor arabinofuranosylcytosine (Ara‐C). The exposure time to TPA required to elicit these changes was found to be short, in the order of 1 hour or less. The kinetics of the increase in E + cells, decrease in the levels of TdT in these cells, or decrease in the ability to proliferate as measured by colony formation were similar with exposure to TPA for 1, 6, 24, or 96 hours. We have examined the effect of antitumor promoter compounds on their ability to block induction of MOLT‐3 cells by TPA. Results indicated that none of these compounds, dexamethasone, antipain, retinoic acid, and L‐1‐tosylamide‐2‐phenylethylchloromethyl ketone (TPCK), was effective in reducing the number of E + cells induced by TPA. Examination of three other leukemic T‐cell lines indicated that, in addition to MOLT‐3, the leukemic T‐cell line Jurkat also responded to TPA, whereas two other leukemic T‐cells lines CCRF‐CEM and CCRF‐HSB‐2 did not. Certain physical and morphological changes were also observed after stimulation of MOLT‐3 cells and Jurkat cells by TPA. We found that, following the addition of TPA, the cell volumes of MOLT‐3 cells decreased from an average of 1150 μm 3 to about 500 μm 3 , whereas those of Jurkat were reduced to about 700 μm 3 from 1100 μm 3 . Electron microscopic studies of these lymphoblasts also revealed that after treatment with TPA the induced cells were generally smaller in size with increase in the density of the nuclear materials and condensation of the chromatin structures.