Transcriptional repression of vimentin gene expression by pyrroline dithiocarbamate during 12-O-tetradecanoylphorbol-13-acetate-dependent differentiation of HL-60 cells.
Vimentin is a member of the intermediate filament family, and the NF-kappaB binding site is located in the human vimentin promoter. To gain insight into the role of NF-kappaB in the regulation of the vimentin gene during 12-O-tetradecanoylphorbol-13-acetate (TPA)-dependent differentiation of HL-60 cells, the effect of pyrrolidine dithiocarbamete (PDTC) has been investigated using Northern blot hybridization and DNA mobility shift assay. PDTC inhibited macrophage-like morphologic change of HL-60 cells by TPA. TPA-dependent increase of vimentin mRNA level was decreased in a time- and dose-dependent manner by pretreatment with PDTC. One DNA-protein complex was formed by DNA mobility shift assay when the NF-kappaB or AP-1 binding sites were incubated with nuclear extract prepared from TPA-treated HL-60 cells, but no protein bound in control HL-60 cells without TPA. After PDTC pretreatment, NF-kappaB binding activity vanished but AP-1 binding activity was unchanged. Taken together, these results suggest that NF-kappaB may be an essential transacting factor for transcriptional repression of the vimentin gene by PDTC during TPA-dependent differentiation of HL-60 cells.
Zheng
Meizi
M
Department of Biochemistry, College of Medicine, Chungnam National University, Daejeon 301-747, Korea.
Son
Mee-Young
MY
Park
Chung
C
Park
Jong-Il
JI
Jo
Eun-Kyeong
EK
Yoon
Wan-Hee
WH
Park
Seung-Kiel
SK
Hwang
Byung-Doo
BD
Lim
Kyu
K
D015152:Blotting, Northern; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D048430:Cell Shape / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D024202:Electrophoretic Mobility Shift Assay; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D008858:Microscopy, Phase-Contrast; D016328:NF-kappa B / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011759:Pyrrolidines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013859:Thiocarbamates / Q000494:pharmacology*; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D014746:Vimentin / Q000235:genetics*
D016328:NF-kappa B; D011759:Pyrrolidines; D012333:RNA, Messenger; D013859:Thiocarbamates; D014746:Vimentin; C020972:pyrrolidine dithiocarbamic acid; D013755:Tetradecanoylphorbol Acetate