Akiyama T, Matsumoto J, Ishikawa T, Eguchi G
Differentiation. 1986;33(1):34-44. doi: 10.1111/j.1432-0436.1986.tb00408.x.
We examined the crystallins present in lens-like cell aggregates produced by goldfish erythrophoroma (tumors of integumental erythrophores) cells in vitro using a combination of Sephadex-G-200 gel filtration, one- and two-dimensional sodium-dodecyl-sulfate/polyacryl-amide gel electrophoresis, immunoblotting, and indirect immunofluorescence assays. The two studied neoplastic pigment cell lines, GEM 81 and GEM 218, formed small, spherical, transparent cell aggregates, resembling lentoid bodies, within the cell mounds of monolayer cultures after treatment with dimethylsulfoxide (DMSO) and autologous serum. Partial purification of a water-soluble extract of such lens-like cell aggregates and subsequent immunoblotting using antibodies (polyclonal) against newt whole lens proteins revealed the presence of about 20 unequivocally conjugated peptides with molecular masses of 19-27 kilodaltons. From their antigenicity and their behavior during gel filtration and electrophoresis, most of these peptides were identified as either alpha- or beta-form crystallins. Immunofluorescence microscopy using antibodies to newt whole lens proteins revealed intense fluorescence in the lens-like cell aggregates formed by these erythrophoroma cells, whereas the cell mounds in cultures of the same cell lines that had not been subjected to differentiation induction were almost unlabeled. Thus, goldfish erythrophoroma cells appear to be capable of crystallin production as well as the formation of lens-like cell aggregates upon the induction of differentiation. There is little available information indicating that normal pigment cells are capable of lens formation and crystallin synthesis during vertebrate ontogeny, and thus it is possible that neoplastic transformation of pigment cells is associated with the acquisition of the ability to produce crystallins.
我们运用葡聚糖凝胶G - 200凝胶过滤、一维及二维十二烷基硫酸钠/聚丙烯酰胺凝胶电泳、免疫印迹和间接免疫荧光测定相结合的方法，对金鱼红瘤（体表红色素细胞肿瘤）细胞在体外产生的类晶状体细胞聚集体中存在的晶状体蛋白进行了研究。在用二甲基亚砜（DMSO）和自体血清处理后，所研究的两种肿瘤性色素细胞系GEM 81和GEM 218在单层培养物的细胞堆中形成了小的、球形的、透明的细胞聚集体，类似于晶状体样小体。对这种类晶状体细胞聚集体的水溶性提取物进行部分纯化，随后使用针对蝾螈全晶状体蛋白的抗体（多克隆抗体）进行免疫印迹分析，结果显示存在约20种明确结合的肽段，其分子量在19 - 27千道尔顿之间。从它们的抗原性以及在凝胶过滤和电泳过程中的行为来看，这些肽段中的大多数被鉴定为α - 或β - 晶状体蛋白。使用针对蝾螈全晶状体蛋白的抗体进行免疫荧光显微镜检查发现，这些红瘤细胞形成的类晶状体细胞聚集体中有强烈的荧光，而同一细胞系未经分化诱导的培养物中的细胞堆几乎没有标记。因此，金鱼红瘤细胞似乎能够产生晶状体蛋白，并在分化诱导后形成类晶状体细胞聚集体。几乎没有现有信息表明正常色素细胞在脊椎动物个体发育过程中能够形成晶状体并合成晶状体蛋白，因此色素细胞的肿瘤转化可能与获得产生晶状体蛋白的能力有关。
