Abstract
Tripartite motif protein 32 (TRIM32), an E3 ubiquitin ligase, is a member of the TRIM protein family. However, the underlying function of TRIM32 in gastric cancer (GC) remains unclear. Here, we aimed to explore the function of TRIM32 in GC cells. TRIM32 was induced silencing and overexpression using RNA interference (RNAi) and lentiviral-mediate vector in GC cells, respectively. Moreover, the PI3K/AKT inhibitor LY294002 was used to examine the relationship between TRIM32 and AKT. Quantitative reverse-transcription PCR (qRT-PCR) and western blot were used to determine the mRNA and protein contents. The glucose analog 2-NBDG was used as a fluorescent probe for determining the activity of glucose transport. An annexin V-fluorescein isothiocyanate apoptosis detection kit was used to stain NCI-N87, MKN74, and MKN45 cells. Cell counting kit-8 (CCK-8) assay was used to examine cell proliferation. Our results indicated that TRIM32 was associated with poor overall survival of patients with GC. Moreover, TRIM32 was a proproliferation and antiapoptosis factor and involved in the AKT pathway in GC cells. Furthermore, TRIM32 possibly mediated the metabolism of glycolysis through targeting GLUT1 and HKII in GC cells. Importantly, TRIM32 silencing deeply suppressed the tumorigenicity of GC cells in vivo. Our findings not only enhanced the understanding of the function of TRIM32 but also indicated its potential value as a target in GC treatment.
Highlights
Gastric cancer (GC) is one of the most common tumors, which is the third leading cause of death-related cancer all over the world [1]
Total RNA from cell samples was extracted using TRIzol Reagent (Invitrogen, USA). en, the cDNA synthesis kit (Fermentas, Canada) were used for the RNA reverse transcribed into complementary DNA according to the instructions of the manufacturer. e expression of GAPDH was used to normalize the gene expression and counted using the 2− ΔΔCt method. ree replicates are needed for each analysis. e primers used in this study are presented in Supplementary File 1
The glycolysis-related protein Glucose transporter 1 (GLUT1) and Hexokinase II (HKII) were significantly decreased by the inhibitor LY294002 in oeNC or oeTRIM32 transfected cells. erefore, these results demonstrated that Tripartite motif protein 32 (TRIM32) was involved in the AKT signaling pathway and regulated the glycolysis metabolism through targeting GLUT1 and HKII in GC cells
Summary
Gastric cancer (GC) is one of the most common tumors, which is the third leading cause of death-related cancer all over the world [1]. Due to the high morbidity and mortality rates of GC, the novel effective therapies for GC are urgently needed [2]. Erefore, gaining a deep insight into the molecule mechanism of GC is a critical step for developing novel therapeutic approaches. A previous report has demonstrated that TRIM32 is overexpressed in breast cancer and promotes the proliferation of breast cancer cells [4]. E similar results are obtained in hepatocellular carcinoma cells [5]. TRIM32 inhibits the activity of PI3K/AKT/Foxo signaling in muscle atrophy through improving plakoglobin-PI3K dissociation [6]. The detailed function of TRIM32 in GC cells is less investigated
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
