SNG-M

Overexpressing USP14 Reduced Apoptosis and Promoted EC Cell Migration and Invasion

Ubiquitin-specific protease 14 (USP14), a member of the USP family, which catalyzes ubiquitin cleavage from a range of protein substrates, has been found dysregulated in several cancers. We transfected USP14 overexpression plasmids into HEC-1-B and SNG-M cells to clarify the specific mechanism of USP14 in endometrial carcinoma (EC). First, WB showed that USP14 was highly expressed in EC cells, indicating successful transfection (Figure 1A). The assessment of EC cell proliferation and apoptosis was performed using the colony formation assay and FCM, respectively. The results revealed that upregulating USP14 facilitated the colony-forming ability of HEC-1-B and SNG-M and reduced apoptosis (vs. the vector group, Figure 1B, 1C). Furthermore, apoptosis-related proteins (Bcl-2, Bax, and Cyclin D1) were compared by WB. As a result, overexpressing USP14 dampened the Bax level and enhanced the Bcl-2 and Cyclin D1 profiles (Figure 1D). Subsequently, the Transwell experiment assessed the impact of USP14 on EC cell migratory and invasive capabilities. Interestingly, cell migration and invasion were improved after USP14 overexpression (vs. the vector group) (Figure 1E). In addition, the profiles of EMT-related proteins (E-cadherin and Vimentin) and EMT-related transcription factors (Snail1, ZEB1, and Slug) were compared by WB. USP14 overexpression elevated the profiles of Vimentin, Snail1, ZEB1, and Slug while repressing the E-cadherin level (compared with the vector group, Figure 1F). These findings illustrated that overexpressing USP14 facilitated EC cell migration, invasion, and EMT.