MiR-374b targets GATA3 to promote progression and development of glioblastoma via regulating SEMA3B.

Jianmei Gao, Shuang Bai, Yumin Wang, Shen Zhao,  Zhanbiao He,  Ruijin Wang

Abstract:

In the present study, a series of experiments were conducted to explore the function of miR-374b and the regulatory relationship among miR-374b, GATA3 and SEMA3B in glioma. MiR-374b mimics and inhibitors were employed to regulate miR-374b expression. Besides, qRT-PCR assay was used for detecting the expression level of miR-374b, GATA3 and SEMA3B mRNAs. To verify the targeting relationship between miR-374b and GATA3, dual luciferase analysis was utilized. Moreover, chromatin immunoprecipitation (ChIP) assay was performed to identify the correlation GATA3 with SEMA3B. Furthermore, si1-GATA3, si2-GATA3 and pc-GATA3 were used to regulate GATA3 expression and pc-SEMA3B served for dysregulation the SEMA3B. For assessing the significance of miR374b alone or in co-operation with GATA3 or SEMA3B on cell viability, migration and apoptosis, CCK-8, transwell and FCM assay were also performed. We found that overexpression of miR-374b, which was identified in glioma tissues and cell lines (U251, LN-299 and GOS-3) promoted cell migration and enhanced cell viability but inhibited apoptosis in this study. Furthermore, GATA3 contributed to increase the cell viability and migration and decrease the apoptosis targeted by miR-374b as evidenced by dual luciferase assay. Moreover, GATA3 binding to the promoter of SEMA3B, involved in regulating SEMA3B, was revealed. Further, a series of studies demonstrated that miR-374b targeted GATA3 regulating SEMA3B and resulted in elevation of cell viability and migration but suppressed the apoptosis. However, the promotion effects of miR-374 in glioma process were reversed by co-transfecting pc-GATA3 or pc-SEMA3B. In conclusion, miR-374b promotes glioma process in vitro through suppressing SEMA3B via targeting GATA3. The result of this study provides an important clue to the optimal treatment schedule for glioblastoma.