Abstract
Engelbreth-Holm-Swarm (EHS) tumors produce large amounts of basement membrane (BM) components that are widely used as cell culture substrates mimicking BM functions. To delineate the tissue/organ origin of the tumor and the mechanisms operating in the BM overproduction, a genome-wide expression profile of EHS tumor was analyzed using RIKEN cDNA microarrays containing approximately 40,000 mouse cDNA clones. Expression profiles of F9 embryonal carcinoma cells that produce laminin-1 and other BM components upon differentiation into parietal endoderm-like cells (designated F9-PE) were also analyzed. Hierarchical clustering analysis showed that the gene expression profiles of EHS and F9-PE were the most similar among 49 mouse tissues/organs in the RIKEN Expression Array Database, suggesting that EHS tumor is parietal endoderm-derived. Quantitative PCR analysis confirmed that not only BM components but also the machineries required for efficient production of BM components, such as enzymes involved in post-translational modification and molecular chaperones, were highly expressed in both EHS and F9-PE. Pairs of similar transcription factor isoforms, such as Gata4/Gata6, Sox7/Sox17, and Cited1/Cited2, were also highly expressed in both EHS tumor and F9-PE. Time course analysis of F9 differentiation showed that up-regulation of the transcription factors was associated with those of BM components, suggesting their involvement in parietal endoderm specification and overproduction of the BM components.
Highlights
Engelbreth-Holm-Swarm (EHS) tumors produce large amounts of basement membrane (BM) components that are widely used as cell culture substrates mimicking Basement membranes (BMs) functions
Global Gene Expression Profiles of EHS Tumor and Differentiated F9 Cells—To confirm that the overproduction of BM components by EHS tumor is regulated at the level of gene expression, total RNA from tumor cells was subjected to Northern blotting analysis using cDNAs encoding mouse laminin-1 subunits (␣1, 1, and ␥1) as probes
The present study was primarily aimed at elucidating the origin of EHS tumor, an extract of which has been widely used as a practical source of BM in many cell biological/tissueengineering studies, and the molecular basis of its overproduction of BM
Summary
Engelbreth-Holm-Swarm (EHS) tumors produce large amounts of basement membrane (BM) components that are widely used as cell culture substrates mimicking BM functions. To further elucidate the regulatory mechanisms of BM production, we analyzed the gene expression profiles of murine F9 embryocarcinoma cells that differentiate into parietal endoderm-like cells (designated F9-PE) and produce large amounts of BM components upon treatment with all-trans-retinoic acid (RA) and dibutyryl cAMP (Bt2cAMP) [16, 17]. Utilizing these approaches, we characterized EHS tumor as a parietal yolk sac-
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