Evaluation of Apoptotic Effect of Protein Fusion, Enterocin A, R-Type Pyocin, Lactocin, and Specific Ligand on Gastric Cancer Cell Line (AGS)

Background and Aim: Conventional anti-cancer treatments, such as surgery, chemotherapy, radiation therapy, etc., are linked to antimicrobial resistance (AMR) and have negative effects on healthy tissues. The flow cytometry technique was used in this study to evaluate the effectiveness of a modified anti-cancer peptide in treating the AGS gastric cancer cell line. Materials and Methods: This study used bioinformatics software to determine the EntA-PynR-Lac gene sequence. The engineered E. coli BL21 bacterium was used to express recombinant proteins derived from the synthesized fusion gene cloned into the pET22b expression vector. A recombinant protein was designed, and its three-dimensional structure and stability were evaluated. Using Western blotting and nickel column chromatography, the recombinant protein was confirmed and purified. In addition, the MTT technique was used to determine the cell lethality of various concentrations of the recombinant protein in AGS cells. Flow cytometry was used to determine the level of apoptosis in AGS cells treated with the desired protein concentrations. Results: According to the results of the MTT test, the 80µg/mL concentration of recombinant protein showed significant cytotoxic effects on the AGS cell line. Furthermore, using flow cytometry, it was found that cancer cells treated with this concentration of recombinant protein exhibited higher rates of total apoptosis than untreated cancer cells. Conclusion: Inhibition of proliferation and growth of gastric cancer cells by this recombinant protein has been shown to be effective. The anti-apoptotic characteristics of the fusion protein make it useful for both therapeutic and preventative purposes when treating cancer cells.