Expression and subcellular redistribution of Bax during TGF-beta1-induced programmed cell death of HC11 mouse mammary epithelial cells.
The effect of TGF-beta1, an auto/paracrine antiproliferative and apoptogenic factor on Bax transcript level (RT-PCR), subcellular distribution of Bax protein (immunoelectron microscopy), Bcl-2 protein level and apoptotic cell number (flow cytometry with FITC-conjugated monoclonal anti-Bcl-2 antibody and DNA stained with DAPI) in HC11 mouse mammary epithelial cells was examined. TGF-beta1 increased Bax transcript level (evaluated by Bax mRNA/GAPDH mRNA ratio) and stimulated Bax protein movement from cytosol to organellar membranes, mainly mitochondrial, during 60 min. The new observation is the presence of Bax on channel membranes of Golgi apparatus and translocation of Bax from cytosol to the fibrous nucleoplasm via nuclear envelope pores (especially after 120 min. of cell exposure to TGF-beta1). Prolactin protected HC11 cells against TGF-beta1-induced PCD, which could occur at two levels: 1) TGF-beta1 expression, through the decrease of TGF-beta1 transcript content, and 2) Bax/Bcl-2 checkpoint, through down-regulation of Bax and up-regulation of Bcl-2. In conclusion, Bax and Bcl-2 proteins are implicated in the mechanism of TGF-beta1-induced PCD and antiapoptotic action of prolactin in HC11 mouse mammary epithelial cells. The activation of transcription and redistribution of Bax from cytosol to organellar membranes and nucleus constitute the early events in the cellular response to TGF-beta1.
Motyl
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Dept. of Animal Physiology, Faculty of Veterinary Medicine, Warsaw Agricultural University, Poland. ploszaj@alpha.sggw.waw.pl
Gajkowska
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B
Płoszaj
T
T
Wareski
P
P
Skierski
J
J
Zimowska
W
W
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count / Q000187:drug effects; D002460:Cell Line; D004815:Epidermal Growth Factor / Q000494:pharmacology; D051379:Mice; D016253:Microscopy, Immunoelectron; D011388:Prolactin / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D051028:bcl-2-Associated X Protein
C494827:Bax protein, mouse; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D051028:bcl-2-Associated X Protein; D004815:Epidermal Growth Factor; D011388:Prolactin