Effects of mitochondrial fission inhibitors on the development of placental trophoblast cells

Objective: The first aim of the study was to perform syncytialization in Bewo and Swan71 cells and to demonstrate syncytialization with biomarkers such as β-hCG and e-cadherin. In the following stages, we aimed to determine the role of mitochondrial fission in syncytialization and to determine to what extent mitochondrial fission inhibitors affect syncytialization. Materials and Methods: In the study, the Bewo and Swan71 system was intervened freely with 20 uM, 50 uM and 100 uM forskolin for 24, 48 and 72 hours. β-hCG and e-cadherin biomarkers were detected in the cells by western blot technique, and the forskolin dose and time for syncytialization were determined. In the following stages, the appropriate dose and time for syncytialization were determined and treated with mdivi-1, a mitochondrial fission inhibitor, at 2 different doses of 50 uM and 100 uM for 3 hours. By monitoring β-hCG and e-cadherin biomarkers detected by western blot technique in those treated and administered with mdivi-1, syncytialization and changes due to mdivi-1 were detected. Results: When the dose of forskolin required for syncytialization was optimized in the study, a statistically significant decrease in e-cadherin expression was observed in the 50 uM and 100 uM groups compared to the K group in the 72h forskolin-treated groups for Bewo cells. In Swan71 cells, protein expression of both β-hcG and e-cadherin biomarkers for syncytialization were not detected. When 72h 100 uM forskolin treated and untreated groups were compared among themselves, there was a statistically insignificant decrease in e-cadherin expression after 50 uM mdivi-1 treatment compared to K group and a statistically insignificant increase in e-cadherin expression after 100 uM mdivi-1 treatment compared to K group. Conclusion: In the first part of the study, syncytialization could be demonstrated with the e-cadherin biomarker in the bewo cell line but not with the β-hCG biomarker. However, syncytialization could not be demonstrated in the swan71 cell line with both β-hCG and e-cadherin biomarker. This suggests that the swan71 cell line does not perform syncytialization, while the bewo cell line does not express sufficient levels of β-hCG protein as in tissues. In the second part of the study, although the results were not significant, the observed increase in e-cadherin expression as a result of the inhibition of mitochondrial fission with mdivi-1 inhibitor suggests that syncytialization does not occur and that mitochondrial differentiation is important for syncytialization. Key Words: Cytotrophoblast, Mdivi-1, Mitochondrial fission, Placenta, Syncytialization, Syncytiotrophoblast.