Fusion of fibroblasts with differentiated and nondifferentiated leukemia cells resulting in blockage of DNA synthesis.

Abstract:

:We have investigated the regulation of DNA synthesis in the heterokaryons of HL60 human myelomonocytic leukemia cells and NIH3T3 mouse fibroblasts to examine if the differentiated leukemia cells contained a replication inhibiting activity. Cell fusions were performed either by exposing a suspension of mixed cells to an electric pulse or by the polyethylene glycol method. To identify the origin of the nuclei in a heterokaryon, one set of partner cells was prelabeled with [3H]thymidine before fusion. DNA synthetic activity after fusion was then revealed immunohistochemically by bromodeoxyuridine incorporation. DNA synthesis in the nuclei of 3T3 was inhibited in the heterokaryons of 3T3 and in either one of the two differentiated forms of HL60, i.e., the macrophage-like or the granulocyte-like. The result supports that a negative regulator of DNA synthesis exists in the differentiated HL60. Surprisingly, we have also found that DNA synthesis was inhibited in the nuclei of both 3T3 and nondifferentiated, proliferating HL60 when these two cells were fused. When unfused, proliferating cells were eliminated with cytosine arabinoside; these nonreplicating heterokaryons survived for at least 5 days, and 15% of them showed alpha-naphthylacetate esterase activity, a trait of the macrophage differentiation. The blockage of DNA synthesis in both partner nuclei was also observed in the heterokaryons of NIH3T3 cells and nondifferentiated human promonocytic leukemia cells U937, and in nondifferentiated HL60 and human diploid fibroblasts WI38. However, this effect was not found in the heterokaryons of NIH3T3 cells and human B lymphoma WI-729-HF2 cells. This is the first demonstration of the inhibition of DNA synthesis upon fusion of two proliferating cells.

journal_name

Dev Biol

journal_title

Developmental biology

authors

Prudovsky IA,Tsong TY

doi

10.1016/0012-1606(91)90417-2

subject

Has Abstract

pub_date

1991-04-01 00:00:00

pages

232-9

issue

2

eissn

0012-1606

issn

1095-564X

pii

0012-1606(91)90417-2

journal_volume

144

pub_type

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