1.Effect of MSI2 on proliferation and apoptosis imbalance of KGN cells induced by DHT
Youwen Du ; Haitao Shi ; Shuailong Han ; Shumin Zhang ; Chaoqi Guan ; Tianjuan Wang ; Linxin Pan
Acta Universitatis Medicinalis Anhui 2023;58(3):464-470
Objective:
To study the effect of Musashi-2 ( MSI2 ) overexpression on the imbalance of proliferation and apoptosis of human ovarian granulosa cell line (KGN) induced by dihydrotestosterone (DHT) .
Methods:
The gene expression profiles of human ovarian granulosa cells ( GCs) in primary culture were statistically analyzed to screen the differentially expressed genes.pcDNA3. 1-MSI2 eukaryotic expression plasmid was constructed and transiently transfected into the KGN cells,and the overexpression effect of MSI2 was detected by Quantitative Real-time PCR (RT-qPCR) and Western blot.After overexpressing MSI2 in DHT induced KGN cells,MTT colorimetry and Edu staining were used to detect the proliferation of cells in each group,and flow cytometry ( FCM) was further used to detect the apoptosis of cells in each group.
Results:
The mRNA expression level of MSI2 gradually decreased during the primary culture of human ovarian GCs.And pcDNA3. 1-MSI2 was successfully constructed and transfected into KGN cells to improve the mRNA and protein expression levels of MSI2.Then MTT,EdU and FCM results showed that compared with the blank group,DHT induction could significantly reduce the proliferation rate and increase the apoptosis rate of KGN cells (P <0. 05) .However,after MSI2 overexpression,the proliferation rate of KGN cells increased and the apoptosis rate decreased (P <0. 05) ,which were close to the blank group.
Conclusion
Overexpression of MSI2 can effectively alleviate the imbalance of proliferation and apoptosis of KGN cells induced by DHT,indicating that MSI2 plays an important role in GCs growth and follicle development.