Secretion of phospholipid transfer protein by human hepatoma cell line, Hep G2, is enhanced by sodium butyrate.
Hep G2 cells were used to study the synthesis and secretion of phospholipid transfer protein (PLTP). Upon incubation of the cells at confluence with serum-free Dulbecco's modified Eagle's medium (DMEM), phosphatidylcholine (PC) transfer activity was found to accumulate in the culture media. The PC transfer activity in the media was effectively inhibited by rabbit anti-human PLTP immunoglobulin (Ig)G, thus indicating that the PC transfer activity was due to secreted PLTP. The molecular weight of Hep G2 PLTP was approximately 78 kDa by Western blot analysis, in agreement with the molecular weight obtained for purified human plasma PLTP. The PLTP secreted by Hep G2 also possessed an HDL conversion activity similar to that of human plasma PLTP. The addition of butyrate to the cell culture media resulted in a marked increase in the secretion of PLTP. After 24 h incubation with 4 mmol/L sodium butyrate, a more than twofold increase (P < 0.01) of PC transfer activity in the cell-conditioned media was obtained. The dose-dependent increase in the PC transfer activity in the media upon butyrate treatment was well correlated (r = 0.80, P < 0.01) with that of PLTP mass as determined by immuno-slot blot analysis of cell-conditioned media. The increased secretion of PLTP by Hep G2 treated with sodium butyrate was accompanied by a greater increase in the level of PLTP mRNA in the cells as determined by ribonuclease protection assay. In the presence of 4 mmol/L sodium butyrate, a fourfold increase (P < 0. 01) in mRNA level was obtained at 24 h. No stabilizing effect of butyrate on PLTP mRNA was apparent upon treatment of the cultured cells with the RNA synthesis inhibitor, actinomycin D. Thus, the up-regulatory effect of butyrate on PLTP gene expression seemed to have occurred at the transcriptional level.
Guo
Z
Z
The Burnsides Research Laboratory, Department of Food Science & Human Nutrition and Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801, USA.
Yuan
C
C
Wei-Lavery
T
T
Fang
Y
Y
Garvin
R A
RA
Nishida
H I
HI
Nishida
T
T
D000818:Animals; D015153:Blotting, Western; D020148:Butyric Acid / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006633:Histamine Antagonists / Q000494:pharmacology*; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D049294:Phospholipid Transfer Proteins; D012333:RNA, Messenger / Q000187:drug effects; D011817:Rabbits; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation
D002352:Carrier Proteins; D006633:Histamine Antagonists; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D049294:Phospholipid Transfer Proteins; D012333:RNA, Messenger; D020148:Butyric Acid