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Updated: Jun 4, 2026
Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets
Published on: November 22, 2024
[Screening drugs for regulating tissue factor gene expression]
Yan Yang1, Fan-Zhi Yan, Jin-Song Yan
1Department of Hematology, Dalian Medical University Second Hospital, Dalian 116027, Liaoning Province, China.
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.
Insights
Researchers developed a stable cell line to screen drugs regulating tissue factor (TF) gene expression. Bortezomib (BTZ) activated TF transcription, while CDA-II suppressed it, identifying novel drug candidates.
Area of Science:
- Molecular Biology
- Gene Regulation
- Drug Discovery
Context:
- Tissue factor (TF) plays a critical role in hemostasis and thrombosis.
- Understanding TF gene regulation is crucial for developing targeted therapies.
- Existing methods for screening TF-regulating drugs are limited.
Purpose:
- To establish a stable U937 cell line expressing luciferase under the control of the TF promoter.
- To screen for novel drugs that modulate TF gene expression.
- To identify the specific TF promoter region responsive to drug treatment.
Summary:
- Stable U937 cell lines with TF promoter-luciferase constructs were created.
- Bortezomib (BTZ) at 5 nmol/L was found to activate TF transcription.
- CDA-II at 1 mg/ml was found to suppress TF transcription.
- The critical regulatory region for BTZ and CDA-II activity was mapped to -201 to 0 bp of the TF promoter.
Impact:
- Successful establishment of a novel cell line for TF research.
- Identification of BTZ and CDA-II as potential modulators of TF expression.
- Provides a foundation for further investigation into the molecular mechanisms of TF regulation and drug action.
