Related Experiment Videos
Hydrocortisone-induced accumulation of fibronectin mRNA and cell surface-associated fibronectin
1Frauenhofer Institut für Toxikologie, Hannover, Federal Republic of Germany.
Abstract:
A Morris hepatoma 7777-derived cell line, DTH-3, was used to study the control of fibronectin gene expression. In cultures of DTH-3 cells in conventional medium supplemented with serum or in chemically defined MX-83 medium supplemented with insulin no cell surface fibronectin was detectable by indirect immunofluorescence techniques using specific polyclonal antibodies. By Northern blot hybridization analysis a dose- and time-dependent accumulation of 8 kb fibronectin mRNA in response to hydrocortisone treatment was demonstrated. Furthermore, 24 h after addition of hydrocortisone an extensive fibrillar fibronectin network was established. The results suggest that the hydrocortisone-dependent induction of fibronectin production might, at least in part, be controlled at the transcriptional level.
Insights
Hydrocortisone treatment increased fibronectin mRNA and protein in DTH-3 cells, suggesting transcriptional control. This study investigates fibronectin gene expression regulation.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Fibronectin is a crucial extracellular matrix protein involved in cell adhesion and migration.
- Understanding fibronectin gene expression is vital for research in cell biology and disease.
- The DTH-3 cell line, derived from Morris hepatoma 7777, offers a model for studying gene regulation.
Purpose of the Study:
- To investigate the control mechanisms of fibronectin gene expression in DTH-3 cells.
- To determine the effect of hydrocortisone on fibronectin production and mRNA levels.
- To elucidate whether hydrocortisone influences fibronectin at the transcriptional or post-transcriptional level.
Main Methods:
- Utilized indirect immunofluorescence to detect cell surface fibronectin.
- Employed Northern blot hybridization to analyze fibronectin mRNA accumulation.
- Quantified fibronectin network formation following hydrocortisone treatment.
Main Results:
- No cell surface fibronectin was detected in DTH-3 cells under standard or insulin-supplemented conditions.
- Hydrocortisone treatment induced a dose- and time-dependent accumulation of fibronectin mRNA.
- An extensive fibrillar fibronectin network formed 24 hours post-hydrocortisone addition.
Conclusions:
- Hydrocortisone significantly upregulates fibronectin gene expression in DTH-3 cells.
- The induction of fibronectin production by hydrocortisone appears to be regulated at the transcriptional level.
- These findings provide insights into the hormonal control of extracellular matrix protein synthesis.