title: [Screening drugs for regulating tissue factor gene expression].
abstract: This study was purposed to screen the drugs for regulating tissue factor (TF) gene expression through establishing stable cell line with luciferase gene having TF promoter transcription activity, so as to provide the basis for further studying the molecular mechanism of screened drugs. A series of luciferase reporter gene plasmids under control of 5'-truncated TF promoter (including -2174 bp - +128 bp, -684 bp - +128 bp, -247 bp - +128 bp and -201 bp - +128 bp) were constructed. The above plasmids were separately electroporated into U937 cells to establish stably transfected sublines. The function of stable cell line was testified by treatment with ATRA, the luciferase gene activity was analyzed by treating established cell line with bortezomib (BTZ) and CDA-II, and drugs for regulating TF gene expression were screened. The results indicated that the BTZ of 5 nmol/L could activate TF gene transcription activity, up-regulate the expression level of TF transcripts; CDA-II of 1 mg/ml could suppress TF gene transcription activity, down-regulate the expression level of TF transcripts. The functional analysis of TF promoter transcription revealed that the region of regulating TF promoter transcription activity by BTZ and CDA-II was between -201 to 0 bp. It is concluded that stable cell line U937 expressing luciferase activity of TF promoters is established, the novel drugs regulating TF gene expression are screened out by means of this established cell line. This study provides basis for screening the new drugs and further studying their molecular mechanisms.
authors.affiliation: Department of Hematology, Dalian Medical University Second Hospital, Dalian 116027, Liaoning Province, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Fan-Zhi
authors.initials: FZ
authors.lastname: Yan
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Yan
authors.forename: Jia
authors.initials: J
authors.lastname: Zhao
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Li
authors.forename: Xue-Yu
authors.initials: XY
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Jin
authors.forename: Shu-Mei
authors.initials: SM
authors.lastname: Rao
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D004354:Drug Screening Assays, Antitumor; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011719:Pyrazines / Q000494:pharmacology*; D013925:Thromboplastin / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation; D020298:U937 Cells
chemical_list: D000970:Antineoplastic Agents; D001897:Boronic Acids; D011719:Pyrazines; D014157:Transcription Factors; D000069286:Bortezomib; D013925:Thromboplastin