Structural and functional optimization of glycoprotein-enzymes for targeted biocatalysis in oral squamous cell carcinoma.

The efficacy of optimized glycoproteinenzymes as a novel therapeutic approach for oral squamous cell carcinoma (OSCC) was tested in this study. The stability and viability of SCC-25 and HN4 operating-system cell lines were characterized. Both lines were confirmed to have a spindle-like morphology for SCC-25, while HN4 cells exhibited cobblestone-like clusters. Viability decreased with time for cell clones SCC-25 was 95 % and 80 % after five days, while HN4 was 94 % and 79 %. Enzyme 1, expression in E. coli and Pichia pastoris to high purity recombinant glycoprotein-enzymes. Activities of these enzymes varied equally among experimental conditions. The enzyme showed an activity of 18 units at Condition D as active max, Enzyme 2 retraced 16 units, and Enzyme 3 reached this point in the same condition. Differences in activity between different conditions were also found in various experimental conditions. In therapeutic assessments, glycoprotein-enzyme treatment lowered OSCC cell viability with IC50 values of 10-15 g/ml. Successful cellular localization could be detected primarily in the cytoplasm and nucleus of live animal tissue following treatment with those therapies. In preclinical xenograft models, treatment resulted in a 40-50 % reduction in tumour volume and growth rates, with treated tumours displaying a 60 % decrease in Ki-67, a 50 % reduction in Bcl-2, and a 70 % increase in cleaved caspase-3. Additionally, the Bax/Bcl-2 ratio increased by 80 %, and CD31 staining revealed a 40 % reduction in microvessel density. These results suggest that optimized glycoprotein enzyme therapy effectively inhibits tumour growth, induces apoptosis and reduces angiogenesis, thus laying a solid foundation for its application in clinical therapy of OSCC.