The stability of cytokeratin 18 in human liver cells during colchicine-induced microtubule disruption.
The cytoskeleton plays important roles in cell function and is therefore implicated in the pathogenesis of many human liver diseases, including malignant tumors. The stability of cytokeratin proteins during tumor transformation in human hepatocellular carcinoma has been studied with a molecular approach previously. The results demonstrate that the cytokeratin is modulated in human hepatocellular carcinoma. Besides this, three low molecular weight cytokeratin molecules (named HCC CK) are found. This indicates that these HCC CKs have undergone modulation from the human hepatocyte cytokeratin 18. We also checked the cytokeratin profile of the human hepatoma cell line PLC/PRF/5 with the same methods to ensure the HCC CK molecules are produced by modulation but not protein degradation. The stability of cytokeratin molecules was studied by a different approach. The cytokeratin compositions of human liver cells (Chang cell line) were analysed under the effects of microtubule-disrupting drug (colchicine) by SDS-PAGE, Western blot, immunoprecipitation using a commercially available monoclonal anti-cytokeratin 18 antibody and immunofluorescent staining. Within 1 h of treatment, the microtubule began to collapse and the filamentous structure was shortening. The microtubule had almost collapsed and became fragmented to form a lattice-like network after 24 h of treatment. The cytokeratin was modulated after long-term (24 h) treatment of colchicine, and the molecular weight became 14 kD and the antigenicity was lost. The stability of cytokeratin molecules was related to the intact microtubule network, after disruption of the microtubule the cytokeratin would be modulated. The intact microtubule network was a stabilizing factor of cytokeratin 18 in human liver cells.
Department of Pathology, School of Medicine, China Medical College, 91 Hsueh-Shih Road, Taichung, Taiwan.
Y
Y
Liu
B
B
Su
R
R
Pei
C
C
Yeh
K
K
Yeh
K
K
Ying Lee
Y
Y
Hsu
C
C
Ho
Y
Y
Lai
D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000139:chemically induced / Q000473:pathology; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced / Q000473:pathology; D003078:Colchicine / Q000633:toxicity*; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007633:Keratins / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008113:Liver Neoplasms / Q000139:chemically induced / Q000473:pathology; D008858:Microscopy, Phase-Contrast; D008870:Microtubules / Q000187:drug effects* / Q000648:ultrastructure; D011233:Precipitin Tests; D013997:Time Factors; D014407:Tumor Cells, Cultured
D007633:Keratins; D003078:Colchicine