Hepatocyte-derived cultured cells with unusual cytoplasmic keratin-rich spheroid bodies

Journal Article · Tue Nov 01 04:00:00 EDT 2011 · Experimental Cell Research

DOI:https://doi.org/10.1016/J.YEXCR.2011.08.018· OSTI ID:22212184

Delavalle, Pierre-Yves; Alsaleh, Khaled; Pillez, Andre; Cocquerel, Laurence ^[1]; Allet, Cecile ^[2]; Dumont, Patrick ^[3]; CNRS UMR 8161, F-59021 Lille ^[4]; Loyens, Anne ^[2]; Leteurtre, Emmanuelle ^[5]; Omary, M. Bishr ^[6]; Dubuisson, Jean; Rouille, Yves ^[1]; Wychowski, Czeslaw ^[1]

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1. INSERM U1019, CNRS UMR 8204, CIIL, F-59021 Lille (France)
2. INSERM U837-JPARC, 59045 Lille (France)
3. Universite Lille Nord de France, F-59000 Lille (France)
4. France
5. Service d'Anatomie et de Cytologie Pathologiques, Centre de Biologie Pathologie, CHRU de Lille, Avenue Oscar-Lambret, Lille cedex (France)
6. Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America (United States)

Cytoplasmic inclusions are found in a variety of diseases that are characteristic morphological features of several hepatic, muscular and neurodegenerative disorders. They display a predominantly filamentous ultrastructure that is also observed in malignant rhabdoid tumor (MRT). A cellular clone containing an intracytoplasmic body was isolated from hepatocyte cell culture, and in the present study we examined whether this body might be related or not to Mallory-Denk body (MDB), a well characterized intracytoplasmic inclusion, or whether this cellular clone was constituted by malignant rhabdoid tumor cells. The intracytoplasmic body was observed in electron microscopy (EM), confocal immunofluorescence microscopy and several proteins involved in the formation of its structure were identified. Using light microscopy, a spheroid body (SB) described as a single regular-shaped cytoplasmic body was observed in cells. During cytokinesis, the SB was disassembled and reassembled in a way to reconstitute a unique SB in each progeny cell. EM examination revealed that the SB was not surrounded by a limiting membrane. However, cytoplasmic filaments were concentrated in a whorled array. These proteins were identified as keratins 8 and 18 (K8/K18), which formed the central core of the SB surrounded by a vimentin cage-like structure. This structure was not related to Mallory-Denk body or aggresome since no aggregated proteins were located in SB. Moreover, the structure of SB was not due to mutations in the primary sequence of K8/K18 and vimentin since no difference was observed in the mRNA sequence of their genes, isolated from Huh-7 and Huh-7w7.3 cells. These data suggested that cellular factor(s) could be responsible for the SB formation process. Aggregates of K18 were relocated in the SB when a mutant of K18 inducing disruption of K8/K18 IF network was expressed in the cellular clone. Furthermore, the INI1 protein, a remodeling-chromatin factor deficient in rhabdoid cells, which contain a spheroid perinuclear inclusion body, was found in our cellular clone. In conclusion, our data suggest that Huh-7w7.3 cells constitute an excellent model for determining the cellular factor(s) involved in the process of spheroid perinuclear body formation.

• OSTI ID: 22212184
• Journal Information: Experimental Cell Research, Journal Name: Experimental Cell Research Journal Issue: 18 Vol. 317; ISSN 0014-4827; ISSN ECREAL
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
CELL CULTURES
CHROMATIN
ELECTRON MICROSCOPY
FLUORESCENCE
KERATIN
LIVER
LUCIFERASE
MESSENGER-RNA
NEOPLASMS
SPHEROIDS
TUMOR CELLS