A two-dimensional model of cell movement. Well differentiated human rectal adenocarcinoma cells move as coherent sheets upon TPA stimulation.
We previously found that 12-O-tetradecanoylphorbol-13-acetate (TPA)-enhanced invasion of Matrigel was associated with augmentation of cell motility but not with metalloproteinase activity in a highly metastatic variant (L-10) of human rectal adenocarcinoma cell line RCM-1. In the present study, with a two-dimensional cell motility assay, we investigated morphology of TPA-induced motility and biochemical pathways that may be involved in the induction of such a motile response to TPA. TPA induced active cell locomotion in L-10 cells with characteristic morphology: the cells moved outwards from the cell islands mainly as a localized coherent sheet of cells with few single moved out cells, but not cell proliferation. The front cells showed locomotor morphologies with front-tail polarity and well-spread leading lamella. Thus, this TPA-induced L-10 cell spreading and motility system seems to be a good model to investigate how well-differentiated adenocarcinoma cells move as cohesive cell nests. Agents which selectively modulate the adenylate cyclase or G protein-related pathways, e.g., 2',5'-dideoxyadenosine and pertussis toxin, had negligible effect upon motility. In contrast, the membrane-permeable synthetic diacylglycerol 1-oleoyl-2-acetyl-glycerol, which has been reported to activate protein kinase C (PKC) directly, could induce cell spreading and motility. Unexpectedly, PKC inhibitors staurosporine and H-7 enhanced TPA-induced cell spreading and motility. Staurosporine itself could induce cell spreading and motility. Taken together, these observations suggested possible involvement of PKC in TPA-induced L-10 cell spreading and motility and that staurosporine might have PKC agonist effect on induction of the spreading and motility.
Nabeshima
K
K
Department of Pathology, Miyazaki Medical College, Japan.
Komada
N
N
Inoue
T
T
Koono
M
M
D000230:Adenocarcinoma / Q000473:pathology*; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D011493:Protein Kinase C / Q000378:metabolism; D012004:Rectal Neoplasms / Q000473:pathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D004075:Diglycerides; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate