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Expression of coagulation factor V gene by normal adult human hepatocytes in primary culture
Author(s) -
Mazzorana Marlene,
Baffet Georges,
Kneip Bernard,
Launois Bernard,
GuguenGuillouzo Christiane
Publication year - 1991
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1991.tb04421.x
Subject(s) - northern blot , fibrinogen , albumin , microbiology and biotechnology , biology , western blot , hep g2 , messenger rna , immunoprecipitation , gene , factor v , secretion , coagulation , primary culture , endocrinology , medicine , cell culture , biochemistry , genetics , thrombosis
Summary Normal human adult hepatocytes were examined for their ability to synthesize and secrete factor V using primary culture. The culture medium contained both factor V and factor Va as determined by bioassay and activation experiments. Immunoprecipitation of newly synthesized labelled factor V showed the presence of both native factor V (m.w. 330000) and two fragments of respective molecular weight 3 and 265000. Northern blot analysis revealed the presence of a single 7 kb factor V mRNA in cultured human hepatocytes as in liver biopsies, together with fibrinogen β and albumin transcripts. Relative levels of factor V, fibrinogen β and albumin mRNAs differed when the cells cultured, suggesting that expression of the three corresponding genes might in part be independently regulated. Furthermore, addition of glucocorticoids enhanced factor V and fibrinogen β mRNA levels 1·6‐ and 5‐fold respectively, but did not significantly increase that of albumin. These results provide evidence that human hepatocytes actively participate in the synthesis of plasma factor V and constitute a valuable model to study the common and specific regulations involved in the control of the expression of this gene in human liver.