Apolipoprotein A-I stimulates placental lactogen expression by human trophoblast cells.
Earlier studies from our laboratory indicated that apolipoprotein A-I (Apo A-I) stimulates the acute release of human placental lactogen (hPL) from trophoblast cells in culture. We have now demonstrated that Apo A-I also causes a secondary increase in hPL release, beginning about 6 h after exposure to Apo A-I, that is blocked by cycloheximide and actinomycin D. Apo A-I also stimulated a dose-dependent increase in hPL promoter activity in JAR cells transfected with a 1.1-kilobase (-1078/2) fragment of the hPL3 promoter coupled to a chloramphenicol acetyltransferase (CAT) reporter gene. Maximal stimulation, 5.2-fold above basal levels, occurred at an Apo A-I concentration of 1.5 mg/ml, which is within the physiological concentration of Apo A-I during pregnancy. 37pA, a synthetic amphipathic peptide that mimics the secondary structure of Apo A-I and stimulates the synthesis and release of hPL, also stimulated a dose-dependent increase in CAT activity, with maximal stimulation comparable to that caused by Apo A-I. In addition, Apo A-I stimulated a modest increase in CAT activity in BeWo choriocarcinoma cells, Chinese hamster ovary cells, and HeLa cells. However, the maximal stimulation of hPL promoter activity in the Chinese hamster ovary and HeLa cells (approximately 2.5-fold above basal levels) was less than that in choriocarcinoma cells, suggesting that trophoblast cell nuclear factors may be necessary for maximal expression of the promoter in response to Apo A-I. Taken together, these results indicate that Apo A-I stimulates hPL gene expression, and that DNA elements in the first 1.1 kilobase of the promoter are sufficient for transactivation by Apo A-I.
Handwerger
S
S
Division of Endocrinology, Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039, USA.
Myers
S
S
Richards
R
R
Richardson
B
B
Turzai
L
L
Moeykins
C
C
Meyer
T
T
Anantharamahiah
G M
GM
D000818:Animals; D016632:Apolipoprotein A-I / Q000494:pharmacology*; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D010928:Placental Lactogen / Q000096:biosynthesis* / Q000235:genetics; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D014327:Trophoblasts / Q000378:metabolism*
D016632:Apolipoprotein A-I; D012333:RNA, Messenger; D010928:Placental Lactogen