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In Vitro Inhibitory Effect of Recombinant Human Calprotectin on Nalm6 Leukemia Cell Line
Samira Charkhizadeh1, Mehdi Imani2, Nematollah Gheibi3
1Department of Hematology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
Background & Purpose:
In evaluating new drugs for the treatment of various types of cancer, investigations have been made to discover a variety of anti-tumor compounds with less side effects on normal cells. Investigations have shown that the heterodimers S100A8 and S100A9 inhibit the enzyme casein kinase 2 and then prevent the activation of the E7 oncoprotein. Therefore, the aim of this study was to evaluate the effect of calprotectin as an antitumor compound on the Nalm6 (B cell precursor leukemia cell line).
Materials & Methods:
Transformation of genes encoding S100A8 and S100A9 human, designed in the pQE32 plasmid, was performed by the thermal shock method into E. coli M15 bacteria. After bacterial growth in LB medium, the expression of two S100A8 and S100A9 subunits, the solubility of the protein by SDS-PAGE method was determined. Finally, the S100A8 / A9 complex was equally placed in the microtube. In the next step, the cytotoxic effects of calprotectin produced on the Nalm6 cell line were evaluated using the wst1 test. Then, the apoptosis in these cells was measured using flow cytometry methods with Annexin-V coloration.
Results:
In the current study, the results showed that the cytotoxic effects of Calprotectin are time and concentration- dependent. Therefore, it can reduce the tumor expression and had a beneficial effect by induced apoptosis in Nalm6 cell line.
Conclusion:
Calprotectin has an anti-tumor effect on the Nalm6 cell line by increasing apoptosis.
Insights
Calprotectin, a compound involving S100A8 and S100A9, demonstrates antitumor effects by inducing apoptosis in Nalm6 leukemia cells. Its efficacy is concentration and time-dependent, offering a potential new avenue for cancer treatment.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Research into novel anti-cancer drugs focuses on compounds with reduced side effects.
- S100A8 and S100A9 heterodimers inhibit casein kinase 2, preventing E7 oncoprotein activation.
Purpose of the Study:
- To evaluate the anti-tumor potential of calprotectin on the Nalm6 B cell precursor leukemia cell line.
Main Methods:
- Genes for human S100A8 and S100A9 were expressed in E. coli.
- Calprotectin's cytotoxic effects on Nalm6 cells were assessed using the WST1 assay.
- Apoptosis induction was measured via Annexin-V staining and flow cytometry.
Main Results:
- Calprotectin exhibited concentration- and time-dependent cytotoxic effects on Nalm6 cells.
- The compound demonstrated a beneficial effect by inducing apoptosis in the Nalm6 cell line.
Conclusions:
- Calprotectin possesses anti-tumor properties against the Nalm6 cell line.
- The anti-tumor mechanism involves the induction of apoptosis.
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