Nabeshima K, Komada N, Inoue T, Koono M
Department of Pathology, Miyazaki Medical College, Japan.
Pathol Res Pract. 1995 Feb;191(1):76-83. doi: 10.1016/S0344-0338(11)80929-5.
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.
我们之前发现，在人直肠腺癌细胞系RCM-1的高转移变体（L-10）中，12-O-十四烷酰佛波醇-13-乙酸酯（TPA）增强的基质胶侵袭与细胞运动性增强有关，但与金属蛋白酶活性无关。在本研究中，我们采用二维细胞运动分析方法，研究了TPA诱导的运动形态以及可能参与这种对TPA运动反应诱导的生化途径。TPA诱导L-10细胞进行活跃的细胞运动，其具有特征性形态：细胞主要以局部连贯的细胞片形式从细胞岛向外移动，很少有单个细胞移出，且不是细胞增殖。前端细胞呈现出具有头尾极性和充分伸展的前缘薄片的运动形态。因此，这种TPA诱导的L-10细胞铺展和运动系统似乎是一个很好的模型，可用于研究高分化腺癌细胞如何作为凝聚的细胞巢移动。选择性调节腺苷酸环化酶或G蛋白相关途径的试剂，如2',5'-二脱氧腺苷和百日咳毒素，对细胞运动性的影响可忽略不计。相反，据报道可直接激活蛋白激酶C（PKC）的膜通透性合成二酰基甘油1-油酰基-2-乙酰基甘油，可诱导细胞铺展和运动。出乎意料的是，PKC抑制剂星形孢菌素和H-7增强了TPA诱导的细胞铺展和运动。星形孢菌素本身可诱导细胞铺展和运动。综上所述，这些观察结果表明PKC可能参与了TPA诱导的L-10细胞铺展和运动，并且星形孢菌素可能对诱导铺展和运动具有PKC激动剂作用。
