Determination of the expression pattern of immune-checkpoint molecules in breast cancer cell line treated by Toxoplasma gondii TLA antigen

Abstract

Considering the discussed anti-tumor effect of Toxoplasma gondii, we decided to evaluate the effect of TLA antigen of Toxoplasma gondii on the cells by examining the expression of genes and miRNAs related to immune control molecules or checkpoint. Let's talk about tumoral breast cancer. Aim: Determining the change in the expression level of immune control molecules and their regulatory micro-RNAs, caused by the effect of Toxoplasma TLA antigen on breast cancer cells. Methods: Cancer tumor cell line after reaching a density above 80% was treated with Toxoplasma TLA antigen. Then, cell line biomarkers including PD-L1, CD80, miR-424 and miR-155 were isolated using ultracentrifugation and confirmed by transmission electron microscopy and immunoblotting. After extracting cellular RNA and finally synthesizing cDNA, the expression levels of genes related to endothelial-mesenchymal transition were checked by quantitative real-time PCR. Results: In this study, the expression levels of 4 biomarkers PD-L1, CD80, miR-424 and miR-155 were compared between two groups treated with Toxoplasma TLA antigen and the control group. The expression level of PD-L1 was significantly lower in the treated group (0.79 versus 1 with P value = 0.01) and the expression level of miR-424 (3.34 versus 1 with P value < 0.001). ) and miR-155 (2.19 vs. 1 with P value = 0.001) was significantly higher in the treated group. In comparing the CD80 expression level between the two groups treated with Toxoplasma TLA antigen and the control group, the level of this biomarker was higher in the treated group; But the difference between the two groups was not statistically significant (1.13 versus 1 with P value = 0.07).