Fibronectin assembly in the crypts of cytokinesis-blocked multilobular cells promotes anchorage-independent growth.
Anchorage-independent growth is a characteristic feature of cancer cells. However, it is unclear whether it represents a cause or a consequence of tumorigenesis. For normal cells, integrin-mediated adhesion is required for completion of the G1 and cytokinesis stages of the cell cycle. This study identified a mechanism that can drive anchorage-independent growth if the G1 checkpoint is suppressed. Cells with defective G1 checkpoint progressed through several rounds of the cell cycle in suspension in spite of uncompleted cytokinesis, thereby forming bi- and multilobular cells. Aurora B and CEP55 were localized to midbodies between the lobes, suggesting that the cytokinesis process reached close to abscission. Integrin-mediated re-attachment of such cells induced cytokinesis completion uncoupled from karyokinesis in most cells. However, a portion of the cells instead lost the constriction and became binucleated. Also, long-term suspension culture in soft agar produced colonies where the cytokinesis block was overcome. This process was fibronectin-dependent since fibronectin-deficient cells did not form colonies unless fibronectin was expressed or exogenously added. While fibronectin normally is not deposited on non-adherent single cells, bi/multilobular cells accumulated fibronectin in the intussusceptions. Based on our data we conclude: 1) Suppression of the G1 checkpoint allows multiple rounds of the cell cycle in detached cells and thereby enables matrix formation on their surface. 2) Uncompleted cytokinesis due to cell detachment resumes if integrin interactions are re-formed, allowing colony formation in soft agar 3) Such delayed cell division can generate binucleated cells, a feature known to cause chromosomal instability.
Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
Rajesh Kumar
RK
Gupta
Staffan
S
Johansson
D000818:Animals; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002455:Cell Division; D045744:Cell Line, Tumor; D048749:Cytokinesis; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008938:Mitosis; D008954:Models, Biological; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
D005353:Fibronectins