Borenfreund E, Bendich A
Lab Invest. 1978 Mar;38(3):295-303.
Epithelioid liver cells were established in culture from rats sacrificed 10 to 18 weeks after the administration of the hepatocarcinogen diethylnitrosamine in their drinking water. After approximately 3 months in vitro, more than half the cells propagated from rats that developed hepatocellular carcinomas had bean-shaped, acentrically displaced nuclei with large juxtanuclear homogeneous appearing areas resembling the hyalin or Mallory bodies in the livers of chronic alcoholics. These abnormalities were not seen in the livers of origin, but were retained in the carcinomas that formed after the cultured cells containing such juxtanuclear hyalin inclusions were inoculated into young rats or nude (i.e., thymusless) mice; these features persisted upon reestablishment and continuous passage of the tumor cells in culture. The cells were further characterized by their karyotypes and their growth properties in liquid media and soft agar. By transmission electron microscopy the hyaline bodies in the culture tumor cells were shown to consist of a disorganized meshwork of filaments. Examination by incubating cells with cytochalasin B and by using antiactin and anti-DNase antisera as indirect immunofluorescence probes also revealed a disturbance in the cytoskeleton.
从饮用含肝癌致癌物二乙基亚硝胺的大鼠体内分离出上皮样肝细胞进行培养，这些大鼠在给药后10至18周被处死。在体外培养约3个月后，从发生肝细胞癌的大鼠中传代的细胞，超过半数具有豆形、偏心移位的细胞核，核旁有大的均质区，类似慢性酒精中毒患者肝脏中的透明样变或马洛里小体。这些异常在原肝脏中未见，但在将含有这种核旁透明样变包涵体的培养细胞接种到幼鼠或裸鼠（即无胸腺小鼠）后形成的癌中保留下来；这些特征在肿瘤细胞在培养中重新建立并连续传代后仍然存在。通过核型分析以及在液体培养基和软琼脂中的生长特性对这些细胞进行了进一步鉴定。通过透射电子显微镜观察发现，培养的肿瘤细胞中的透明样小体由杂乱的细丝网络组成。用细胞松弛素B孵育细胞，并使用抗肌动蛋白和抗脱氧核糖核酸酶抗血清作为间接免疫荧光探针进行检测，也揭示了细胞骨架的紊乱。
