Expression of alpha2-macroglobulin by the interaction between hepatocytes and endothelial cells in coculture.
The interaction between distinct cell types within the liver seems to be important in regulating hepatic function. However, these interactions have not been well characterized because of difficulty in reproducing the hepatic environment in an ex vivo model. In the present study a coculture system of hepatocytes and endothelial cells was established to investigate the communication between parenchymal and nonparenchymal cells. Freshly isolated rat hepatocytes were placed onto a monolayer of primary aortic rat endothelial cells. Analysis of the proteins secreted into the extracellular medium after pulse labeling with radioactive amino acids revealed the presence of a 180,000-apparent molecular weight glycoprotein, BBB-180, which was not detected in the extracellular medium of hepatocytes or endothelial cells when they were cultured separately. This glycoprotein was identified as alpha2-macroglobulin after sequencing of the proteolytic peptides derived from the purified protein. This finding was confirmed by Northern and Western blotting, immunoprecipitation, and RT-PCR. The expression of alpha2-macroglobulin required direct contact between hepatocytes and viable endothelial cells. These findings suggest that endothelial cells modulate hepatocyte gene expression by direct cellular interactions.
Talamini
M A
MA
Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
McCluskey
M P
MP
Buchman
T G
TG
De Maio
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta; D002450:Cell Communication; D002478:Cells, Cultured; D018920:Coculture Techniques; D003545:Cysteine / Q000378:metabolism; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry; D008099:Liver / Q000166:cytology / Q000502:physiology*; D008297:Male; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013462:Sulfur Radioisotopes; D000511:alpha-Macroglobulins / Q000096:biosynthesis* / Q000737:chemistry
D017931:DNA Primers; D006023:Glycoproteins; D013462:Sulfur Radioisotopes; D000511:alpha-Macroglobulins; D008715:Methionine; D003545:Cysteine