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Lactoferrin gene promoter: structural integrity and nonexpression in HL60 cells
J J Johnston1, P Rintels, J Chung
1Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510.
Blood
|June 11, 1992
Summary
Researchers investigated lactoferrin gene expression in neutrophils and acute leukemia. They found the lactoferrin gene structure is normal in HL60 leukemia cells, but it remains untranscribed, suggesting a regulatory defect.
Area of Science:
- Molecular Biology
- Hematology
- Gene Regulation
Background:
- Lactoferrin, an iron-binding protein, is crucial in neutrophils and glandular tissues.
- Understanding lactoferrin gene expression is vital for neutrophils and in conditions like acute leukemia.
Purpose of the Study:
- To investigate lactoferrin gene expression mechanisms in neutrophils.
- To identify defects in lactoferrin expression in acute leukemia, specifically in HL60 cells.
Main Methods:
- Cloning of lactoferrin cDNA and genomic DNA.
- Polymerase chain reaction (PCR), primer extension, and S1 analysis to define mRNA 5' end and promoter region.
- Analysis of HL60 cell DNA and gene transcription using Northern analysis, nuclear run-on, and primer extension assays.
Main Results:
- The 5' end and putative promoter region of the lactoferrin gene are structurally normal in HL60 cells.
- The lactoferrin gene is not transcribed in DMSO-induced HL60 cells, despite phenotypic neutrophil maturation.
- These findings indicate a defect in a transacting regulator of lactoferrin gene expression.
Conclusions:
- The defect in lactoferrin gene expression in HL60 cells is not due to structural abnormalities in the gene itself.
- A deficiency in a transacting factor is likely responsible for the lack of lactoferrin transcription in these leukemia cells.