title: Effects of 17beta-estradiol, progesterone, synthetic progestins, tibolone, and raloxifene on vascular endothelial growth factor and Thrombospondin-1 messenger RNA in breast cancer cells.
abstract: To determine the effect of 17beta-estradiol, raloxifene, progesterone, medroxyprogesterone acetate (MPA), levonorgestrel (LNG), norethindrone (NET), tibolone and tibolone metabolites on vascular endothelial growth factor (VEGF) isoforms 121 and 165 and Thrombospondin-1 (TSp-1) messenger RNA (mRNA) in two breast cancer cell lines, MCF-7 and T47-D. MCF-7 and T47-D cells were cultured to 80% confluence, in vitro. After 24 h incubation in serum-free media, 1.0, 0.1, and 0.01 muM of 17beta-estradiol, raloxifene, raloxifene plus ICI 182780, tibolone, 3alpha-hydroxytibolone, and 3beta-hydroxytibolone were added to MCF-7 cells. Progesterone, MPA, LNG, NET, and Delta(4) tibolone at 1.0, 0.1, and 0.01 muM were added to T47-D cells. The cells plus steroids were incubated for a further 24 h. Total RNA was isolated using TRIZOL and reverse transcriptase-polymerase chain reaction was carried out using primers for VEGF, TSP-1, and cyclophilin, the latter as an internal control. Semiquantitative analysis was performed using 33P-CTP for radioactive labeling during the polymerase chain reaction. 17beta-estradiol, raloxifene, tibolone, 3alpha-hydroxytibolone, and 3beta-hydroxytibolone had no effect on VEGF mRNA in MCF-7 cells. Progesterone, MPA, LNG, and NET increased VEGF mRNA in T47-D cells. Delta(4) tibolone also increased VEGF mRNA but to a lesser extent than the progestogens. Raloxifene increased TSP-1 mRNA, this effect was not reversed by the addition of ICI 182780 to the media. 17beta-estradiol, raloxifene, tibolone and tibolone hydroxy-metabolites had no effect on VEGF mRNA in MCF-7 cells. Progesterone and progestins increased VEGF mRNA in T47-D breast cancer cells. Delta(4) tibolone was less effective than progestogens on this angiogenic gene in the T47-D cells. Raloxifene increased TSP-1. These differential effects may be related to breast cancer growth.
authors.affiliation: Contraceptive Research and Development Program, Clinical Research Center, Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, Virginia 23507, USA. mirkins@yahoo.com
authors.forename: S
authors.initials: S
authors.lastname: Mirkin
authors.forename: B C
authors.initials: BC
authors.lastname: Wong
authors.forename: D F
authors.initials: DF
authors.lastname: Archer
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006728:Hormones / Q000494:pharmacology*; D006801:Humans; D009652:Norpregnenes / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology; D045167:Progesterone Congeners / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism*; D020849:Raloxifene Hydrochloride / Q000494:pharmacology; D019700:Thrombospondin 1 / Q000235:genetics*; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D006728:Hormones; D009652:Norpregnenes; D045167:Progesterone Congeners; D012333:RNA, Messenger; D019700:Thrombospondin 1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D020849:Raloxifene Hydrochloride; D011374:Progesterone; D004958:Estradiol; C027385:tibolone