Fusion Proteins in the Fight against Colorectal Cancer: Assessing the Cytotoxic Potential of Recombinant Nisin-Arginine Deiminase Fusion Protein

Background: A promising strategy in cancer therapy involves the production of fusion proteins, which entail the fusion of two distinct proteins. This study aimed to produce and assess the cytotoxic effects of the Nisin-arginine deiminase (ADI) fusion protein on the SW۴۸۰ cell lines, a common model for studying colorectal cancer (CRC). Materials & Methods: The designed Nisin-ADI gene fragment sequence was sent to Biomatik Company for synthesis in pET-۲۸a vectors between SacI and HindIII restriction enzyme sites. Escherichia coli (E. coli) DH۵α and BL۲۱ were utilized for cloning and protein expression, respectively. The recombinant fusion protein expression was induced by Isopropyl ß-D-۱-thiogalactopyranoside (IPTG) and purified using Ni۲+-nitrilotriacetic acid (۱) resin affinity chromatography. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were conducted to analyze the purified protein. The cytotoxic effect of the purified recombinant fusion protein on SW۴۸۰ and NIH۳T۳ cells, as a control, was evaluated using ۳-[۴,۵-dimethylthiazol-۲-yl]-۲,۵ diphenyl tetrazolium bromide (MTT) assay. Findings: The results of this study showed that the fusion protein had a significant impact on the SW۴۸۰ cell lines. The Half-maximal inhibitory concentration (IC۵۰) of the fusion protein was ۳۰ µg/mL, indicating that it effectively inhibited the growth of cancer cells. However, the fusion protein did not significantly affect the control group. Conclusion: This study provides helpful insights into the potential application of recombinant Nisin-ADI fusion proteins as a potential treatment option for colorectal cancer. The potential for selective targeting of cancer cells is promising as normal cells are unaffected by this fusion protein.

Keywords:

Nisin , Arginine deiminase , Fusion protein , Colorectal cancer , Cytotoxicity