Watanabe H, Chigira M
Int J Oncol. 1992 Sep;1(4):481-7. doi: 10.3892/ijo.1.4.481.
We established new two protein-free culture subclones from murine well-characterized Ehrlich ascites carcinoma and P815 mastocytoma using intermittent protein-free culture performed previously for a protein-free subclone of fibrosarcoma (Gc-4 PF). The Ehrlich protein-free subclone (Ehrlich PF) grew much more slowly than the original cell line and showed a proliferative response to FCS. On the other hand, like Gc-4 PF, the P815 protein-free subclone (P815 PF) showed a similar growth rate to that of the original counterpart. Interestingly the original P815 mastocytoma cells also grew exponentially in protein-free medium. Although the protein-free culture exhibited cells that were more spheroid and spread less in each of these three cell lines, the major structure protein bands demonstrated on SDS-PAGE were virtually identical between the original and protein-free culture cells. In contrast to the structural peptides, the distribution of the secretory peptide differed among the three protein-free culture cell lines, which may reflect their state of differentiation. Growth-inhibiting activities were detected from the supernatant of all three protein-free culture cells, while no protein-free culture cells secreted predominantly growth-stimulating activity into their cultured media. These results suggest that autonomy in tumor cell proliferation may result from the acquisition of the ability to escape from negative control in multicellular organisms, as shown in monads, rather than an acquisition of further response to growth-stimulating control.
我们使用先前为纤维肉瘤的无蛋白亚克隆(Gc-4 PF)进行的间歇性无蛋白培养,从特征明确的小鼠艾氏腹水癌和P815肥大细胞瘤中建立了新的两个无蛋白培养亚克隆。艾氏无蛋白亚克隆(Ehrlich PF)的生长速度比原始细胞系慢得多,并对胎牛血清(FCS)表现出增殖反应。另一方面,与Gc-4 PF一样,P815无蛋白亚克隆(P815 PF)的生长速度与原始对应物相似。有趣的是,原始的P815肥大细胞瘤细胞在无蛋白培养基中也呈指数生长。尽管无蛋白培养显示这三种细胞系中的细胞更呈球形且铺展较少,但在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)上显示的主要结构蛋白条带在原始细胞和无蛋白培养细胞之间几乎相同。与结构肽相反,分泌肽的分布在三种无蛋白培养细胞系中有所不同,这可能反映了它们的分化状态。在所有三种无蛋白培养细胞的上清液中均检测到生长抑制活性,而没有无蛋白培养细胞向其培养基中分泌主要的生长刺激活性。这些结果表明,肿瘤细胞增殖的自主性可能源于获得了在多细胞生物体中逃避负调控的能力,如单细胞生物所示,而不是获得了对生长刺激调控的进一步反应。