A study was made of whether normal and morphologically transformed colonies in the Syrian hamster embryo (SHE) cell transformation assay performed heterologous gap junctional intercellular communication (GJIC). Two compounds, 12-O-tetradecanoylphorbol-13-acetate (TPA) and Na-orthovanadate (vanadate), which induce high frequencies of morphological transformation in SHE cells, have been employed. Three approaches were used to study the possibility of heterologous GJIC. (i) Morphologically transformed colonies (induced by TPA) partially overlapping with normal colonies were selected. Cells in the border area were micro-injected with the fluorescent dye Lucifer yellow to determine whether the dye spread to cells belonging to the other colony. This approach proved to be unsuccessful due to an inability to pinpoint which cells belonged to which colony. (ii) X-irradiated, non-dividing feeder cells are easily recognized by their large size. Feeder cells in contact with normal or TPA-transformed colonies were injected with Lucifer yellow. The dye was found to spread to most of the contacting cells, irrespective of whether they belonged to a normal or morphologically transformed colony. (iii) TPA- and vanadate-exposed colonies were labelled by endocytosis of Lucifer yellow overnight. This resulted in a punctate fluorescent pattern. Unlabelled, previously unexposed cells were seeded onto the dishes and incubated for 3.5-7 h. The ability to perform heterologous GJIC between the newly seeded cells and labelled colony cells was investigated. Both normal and transformed colonies were found to be able to communicate with the newly seeded cells. Thus, the present results indicate that selective communication is not a general property of morphologically transformed SHE cell colonies.
|Number of pages||6|
|Publication status||Published - 1993|
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