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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1998 Jan;89(1):67–75. doi: 10.1111/j.1349-7006.1998.tb00481.x

Putrescine‐stimulated Intracellular Ca2+ Release for Invasiveness of Rat Ascites Hepatoma Cells

Yoshiyuki Ashida 1,†, Akemichi Ueno 1, Yoshihiro Miwa 1, Keiko Miyoshi 1, Hideo Inoue 1,✉
PMCID: PMC5921585  PMID: 9510478

Abstract

Our previous study showed that treatment of highly invasive rat ascites hepatoma (LC‐AH) cells with α‐difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase, decreased both their intracellular level of putrescine and their in vitro invasion of a monolayer of calf pulmonary arterial endothelial (CPAE) cells, and that both these decreases were completely reversed by exogenous putrescine, but not spermidine or spermine. Here we show that all adhering control (DFMO‐untreated) cells migrated beneath CPAE monolayer with morphological change from round to cauliflower‐shaped cells (migratory cells). DFMO treatment increased the number of cells that remained round without migration (nonmigratory cells). Exogenous putrescine, but not spermidine or spermine, induced transformation of all nonmigratory cells to migratory cells with a concomitant increase in their intracellular Ca2+ level, [Ca2+]i. The putrescine‐induced increase in their [Ca2+]i preceded their transformation and these effects of putrescine were not affected by antagonists of the voltage‐gated Ca2+ channel, but were completely suppressed by ryanodine, which also suppressed the invasiveness of the control cells. The DFMO‐induced decreases in both [Ca2+]i and the invasiveness of the cells were restored by thapsigargin, which elevated [Ca2+]i by inhibiting endoplasmic Ca2+‐ATPase, indicating that thapsigargin mimics the effects of putrescine. These results support the idea that putrescine is a cofactor for Ca2+ release through the Ca2+ channel in the endoplasmic reticulum that is inhibited by ryanodine, this release being initiated by cell adhesion and being a prerequisite for tumor cell invasion.

Keywords: Putrescine, Tumor invasion, Difluoromethylornithine, Ryanodine, Thapsigargin

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