Borenfreund E, Higgins P J, Peterson E
J Natl Cancer Inst. 1980 Feb;64(2):323-33. doi: 10.1093/jnci/64.2.323.
The endoskeletal structure of tumor cells with a characteristic cytoplasmic abnormality initiated in rat liver by the in vivo adminisration of the carcinogen diethylnitrosamine was studied in clonal cell lines established and propagated in vitro. The bulk of the cytoplasmic and plasma membrane protein was removed by extraction with Triton X-100, and subsequently the juxtanuclear detergent-insoluble fraction containing filaments of 100-150 A was released into citrate buffer at pH 2.8. Analysis of this fraction by sodium dodecyl sulfate acrylamide gel electrophoresis revealed the pressence of five major proteins that banded with apparent molecular weights of about 66, 57, 52, 48 , and 43 x 10(3), the last of which comigrated with actin. The proteins thus resembled those from intermediate-sized filaments of both the vimentin (57 x 10(3)) and the prekeratin types obtained from various vertebrate cells. They also appeared to be related to the polypeptides of intermediate-sized filaments from Mallory bodies induced by griseofulvin in the livers of mice and to some of the polypeptides seen in isolates of Mallory bodies from human alcoholics. These results indicated that a major component of the carcinogen-induced lesion consisted of intermediate-sized filaments. The possible significance in cell transformation of this stably maintained aggregate of filaments that binds concanavalin A and displaces the nucleus is discussed. The close resemblance of this lesion to that seen in the cells of cirrhotic livers of alcoholics (Mallory's alcoholic hyalin) raises a question regarding the possible oncogenic status of such cells in humans.
通过在体内给予致癌物二乙基亚硝胺在大鼠肝脏中引发具有特征性细胞质异常的肿瘤细胞的内骨骼结构，在体外建立并传代的克隆细胞系中进行了研究。用Triton X-100提取去除大部分细胞质和质膜蛋白，随后将含有100 - 150埃细丝的近核去污剂不溶性部分释放到pH 2.8的柠檬酸盐缓冲液中。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳对该部分进行分析，发现存在五种主要蛋白质，其条带的表观分子量约为66、57、52、48和43×10³，其中最后一种与肌动蛋白共迁移。这些蛋白质类似于从各种脊椎动物细胞获得的波形蛋白（57×10³）和前角蛋白类型的中间丝中的蛋白质。它们似乎也与在小鼠肝脏中由灰黄霉素诱导的马洛里小体的中间丝多肽以及从人类酒精性肝病患者的马洛里小体分离物中看到的一些多肽有关。这些结果表明致癌物诱导的病变的主要成分由中间丝组成。讨论了这种稳定维持的与伴刀豆球蛋白A结合并使细胞核移位的细丝聚集体在细胞转化中的可能意义。这种病变与酒精性肝硬化患者肝脏细胞中所见病变（马洛里酒精透明小体）的密切相似性引发了关于此类细胞在人类中可能的致癌状态的问题。
