Related Experiment Video
Updated: Feb 7, 2026
Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions
Published on: December 20, 2017
The antitumor effect of TAT-DCF1 peptide in glioma cells
Jiao Wang1, Qian Wang1, Fangfang Zhou1
1Laboratory of Molecular Neural Biology, School of Life Sciences, Shanghai University, 99 Shang Da Road, Shanghai 200444, China.
Background:
Glioblastoma is one of the most malignant brain cancer, thus, establishing an effective therapy is paramount. Our previous results indicate that dendritic cell-derived factor (DCF1) is an attractive candidate for therapy against Glioblastoma, since its overexpression in Glioblastoma U251 cells leads to apoptosis. However, the delivery approach limits its clinical application, in this paper, we expressed TAT-DCF1 fusion protein in E.coli in order to surmount its current delivery problems.
Methods:
The coding sequences of the different domains of DCF1 (full length, cytoplasmic, extracellular, 19-amino acid), together with the N-terminal transactivator of transcription (TAT) sequence, were amplified and subcloned into the bacterial expression vector pET30a(+) in order to produce (His)6-tagged fusion proteins. Coomassie blue-stained SDS-PAGE and Western blotting identification showed that purity of the fusion proteins.
Results:
Immunofluorescence and flow cytometry show that U251 cells were efficiently transduced with the fusion proteins. Cell viability, proliferation, and migration assays suggest that the complete TAT-DCF1 fusion protein significantly decreased U251 proliferation and migration. Flow cytometry further reveals that TAT-DCF1 triggered cellular apoptosis.
Conclusions:
In conclusion, these findings suggest that the TAT-DCF1 fusion protein was efficiently transduced into Glioblastoma U251 cells and induced the antitumor effect and support further investigation into specific targeting and side effects of TAT-DCF1 during drug delivery.
Insights
A novel TAT-DCF1 fusion protein effectively targets Glioblastoma cells, inhibiting proliferation and migration while inducing apoptosis. This advancement offers a promising therapeutic strategy for brain cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Glioblastoma is a highly aggressive brain cancer requiring effective therapies.
- Dendritic cell-derived factor (DCF1) shows potential for Glioblastoma treatment by inducing apoptosis.
- Current delivery methods for DCF1 limit its clinical application.
Purpose of the Study:
- To develop an improved delivery system for DCF1 using a fusion protein approach.
- To express and characterize a TAT-DCF1 fusion protein for Glioblastoma therapy.
Main Methods:
- Amplification and subcloning of DCF1 domains and the TAT sequence into a bacterial expression vector.
- Production of (His)6-tagged TAT-DCF1 fusion proteins in E. coli.
- Analysis of protein purity using SDS-PAGE and Western blotting.
Main Results:
- Efficient transduction of Glioblastoma U251 cells with TAT-DCF1 fusion proteins confirmed by immunofluorescence and flow cytometry.
- Significant reduction in U251 cell proliferation and migration observed with TAT-DCF1 treatment.
- TAT-DCF1 induced cellular apoptosis in Glioblastoma U251 cells.
Conclusions:
- The TAT-DCF1 fusion protein demonstrates efficient transduction and antitumor effects in Glioblastoma cells.
- Further research is warranted to explore specific targeting and potential side effects of TAT-DCF1 for drug delivery.
More Related Videos
Related Concept Videos
02:43Peptide Bonds
01:33Peptide Identification Using Tandem Mass Spectrometry
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
01:19Blood Studies for Cardiovascular System II: CRP, Hcy, and Cardiac Natriuretic Peptide Markers
These markers indicate stress or strain on the heart muscle:
Natriuretic Peptides (BNP)
Cardiac myocytes produce these hormones in response to ventricular stretching...
01:07What are Cells?
Basic Characteristics of Cells
A living cell has a plasma membrane, a bilayer of lipids that separates the aqueous solution inside the cell called the cytoplasm from the outside environment.
Furthermore, a living cell possesses genetic information...
02:03What is Cell Signaling?
02:58Chemistry of the Cell
Water
The polarity of the water molecule and its resulting hydrogen bonding makes water a unique substance with special properties that are intimately tied to the processes of life. Life originally evolved in an aqueous environment, and most of an organism’s cellular chemistry and metabolism occur inside the aqueous contents of the cell’s cytoplasm. Special properties of water are its high heat capacity...