Trophoblast-like human choriocarcinoma cells serve as a suitable in vitro model for selective cholesteryl ester uptake from high density lipoproteins.
As human choriocarcinoma cells display many of the biochemical and morphological characteristics reported for in utero invasive trophoblast cells we have studied cholesterol supply from high density lipoproteins (HDL) to these cells. Binding properties of 125I-labeled HDL subclass 3 (HDL3) at 4 degrees C were similar for BeWo, JAr, and Jeg3 choriocarcinoma cell lines while degradation rates at 37 degrees C were highest for BeWo. Calculating the selective cholesteryl ester (CE)-uptake as the difference between specific cell association of [3H]CE-labeled HDL3 and holoparticle association of 125I-labeled HDL3 revealed that in BeWo cells, the selective CE-uptake was slightly lower than holoparticle association. However, the pronounced capacity for specific cell association of [3H]CE-HDL3 and selective [3H]CE-uptake in excess of HDL3-holoparticle association, and cAMP-mediated enhanced cell association of [3H]CE-HDL3 in JAr and Jeg3 suggested the scavenger receptor class B, type I (SR-BI) to be responsible for this pathway. Abundant expression of SR-BI (but not SR-BII, a splice variant of SR-BI) could be observed in JAr and Jeg3 but not in BeWo cells using RT-PCR, Northern and Western blot analysis, and immunocytochemical technique. Adenovirus-mediated overexpression of SR-BI in all three choriocarcinoma cell lines resulted in an enhanced capacity for cell association of [3H]CE-HDL3 (20-fold in BeWo; fivefold in JAr and Jeg3). The fact that exogenous HDL3 remarkably increases proliferation in JAr and Jeg3 supports the notion that selective CE-uptake and subsequent intracellular generation of cholesterol is coupled to cellular growth. From our findings we propose that JAr and Jeg3 cells serve as a suitable in vitro model to study selective CE-supply to human placental cells.
Wadsack
Christian
C
Institute of Medical Biochemistry and Molecular Biology, Karl-Franzens University Graz, Austria.
Hrzenjak
Andelko
A
Hammer
Astrid
A
Hirschmugl
Birgit
B
Levak-Frank
Sanja
S
Desoye
Gernot
G
Sattler
Wolfgang
W
Malle
Ernst
E
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D018955:CD36 Antigens / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016466:CHO Cells; D002788:Cholesterol Esters / Q000378:metabolism*; D002822:Choriocarcinoma / Q000378:metabolism* / Q000473:pathology; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008075:Lipoproteins, HDL / Q000378:metabolism*; D053440:Lipoproteins, HDL3; D008077:Lipoproteins, LDL / Q000378:metabolism*; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; D018613:Microscopy, Confocal; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D018110:Receptors, Lipoprotein / Q000235:genetics / Q000378:metabolism; D051116:Receptors, Scavenger; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051122:Scavenger Receptors, Class B; D012795:Sialoglycoproteins; D014162:Transfection; D014327:Trophoblasts / Q000378:metabolism*
D018955:CD36 Antigens; D002788:Cholesterol Esters; D008075:Lipoproteins, HDL; D053440:Lipoproteins, HDL3; D008077:Lipoproteins, LDL; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; C074299:SCARB2 protein, human; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; D012795:Sialoglycoproteins; D000242:Cyclic AMP