Synergistic anticancer effects of interleukin-21 combined with therapeutic peptides in multiple cancer cells
Muhammad Akram1, Nao Akusa Fujimura1, Saad Tahir1
1Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Background:
Interleukin-21 (IL-21) is a cytokine produced by various cell types, including T cells, natural killer cells, myeloid cells, and B cells, and has a broad range of potential applications in cancer therapy. To improve the therapeutic index, we explored the use of fusion technologies that involved linking other anticancer peptides to the IL-21 gene using specific linkers.
Objectives:
This study aimed to compare the anticancer potential of IL-21 and IL-21 fusion proteins.
Methods:
Antimicrobial peptides possessing anticancer properties were fused with IL-21 gene using a flexible linker (-GGGGS-), and the resulting construct was inserted into the pSecTag2a mammalian expression vector. The cassette was transfected into several cancer cell lines including H1 HeLa, HepG2, MCF-7, MDA-MB-231, HCT-116, HCC-1954, HEK-293, and SF-767. The cytotoxic effects of IL-21 and fusion proteins were evaluated using MTT, Caspase-3, LDH, and scratch assays.
Results:
The IL-21-Tachyplesin I fusion protein had the strongest antiproliferative activity against all tested cancer cells, followed by IL21-LPSBD2 and IL-21. In contrast, IL21-Cop A3, IL21-CSP I-Plus, and IL21-RGD Temporin-Las did not inhibit the viability of cancer cells.
Conclusion:
Fusion technology is a promising therapeutic technique that can be used to enhance the cytotoxicity and antiproliferative activity of anticancer proteins such as IL-21.
Insights
Fusion technology enhances Interleukin-21 (IL-21) anticancer activity. The IL-21-Tachyplesin I fusion protein showed the strongest antiproliferative effects against various cancer cells, highlighting fusion potential in cancer therapy.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Interleukin-21 (IL-21) is a cytokine with potential in cancer therapy.
- Fusion technology was explored to improve IL-21's therapeutic index by linking it with anticancer peptides.
Purpose of the Study:
- To compare the anticancer efficacy of IL-21 and novel IL-21 fusion proteins.
- To evaluate the potential of fusion technology in enhancing anticancer protein activity.
Main Methods:
- Anticancer peptides were fused to the IL-21 gene using a flexible linker.
- Constructs were transfected into multiple cancer cell lines (HeLa, HepG2, MCF-7, etc.).
- Cytotoxicity was assessed using MTT, Caspase-3, LDH, and scratch assays.
Main Results:
- The IL-21-Tachyplesin I fusion protein exhibited the highest antiproliferative activity across all tested cancer cells.
- IL21-LPSBD2 and IL-21 also showed inhibitory effects.
- Other fusion proteins (IL21-Cop A3, IL21-CSP I-Plus, IL21-RGD Temporin-Las) showed no significant impact on cancer cell viability.
Conclusions:
- Fusion technology is a promising strategy for enhancing the anticancer properties of proteins like IL-21.
- Specific fusion constructs, such as IL-21-Tachyplesin I, demonstrate superior therapeutic potential.
- Further research into fusion technologies could lead to improved cancer treatments.
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