Figures & data
Table 1. TRIM58 interference target design results.
Figure 1. TRIM58 expression is significantly reduced in GC tissues and cell lines Paired cancer and normal tissues of 23 patients were collected. (A) TRIM58 mRNA expression was examined by RT-PCR. (B) TRIM58 in malignant and normal tissues was detected by IHC. (C) mRNA and protein expression of TRIM58 in GC cell lines were examined. (D) β-catenin in cancer and normal tissues was detected by IHC. (E) β-catenin expression (nuclear and cytoplasmic) in GC cell lines was detected. (F-G) mRNA and protein expression of C-myc, Cyclin D1, and survivin in GC tumors and cell lines were detected. *P < .05, **P < .01 and ***P < .001 were compared to Paracancer or GES-1.
Figure 2. Overexpression and knockdown of TRIM58 in GC cells by lentiviral infection After lentiviral infection with vector or TRIM58 and shNC or shTRIM58 (shTRIM58-1, shTRIM58-2, and shTRIM58-3) (A-B) the overexpression efficiency of TRIM58 in AGS and HGC27 cells was detected. (C) The knockdown efficiency of shTRIM58-1, shTRIM58-2, and shTRIM58-3 in SNU719 cells was also detected. ***P < .001 was compared to vector or shNC.
Figure 3. Overexpression of TRIM58 inhibits GC cell proliferation via cell-cycle arrest and increase in cell apoptosis AGS, HGC27, or SNU719 cells were infected with shTRIM58 (shTRIM58-1 and TRIM58-2) or TRIM58 lentiviruses. (A) Cell proliferation was assessed by CCK-8 or BrdU-ELISA at 0, 24, 48, and 72 h. (B-C) After 48 h of infection, cell-cycle phase proportions and apoptosis were assessed by flow cytometry. (D) The levels of related proteins (β-catenin, C-myc, Cyclin D1, and survivin) were analyzed by Western blot. **P < .01 and ***P < .001 were compared to vector or shNC.
Figure 4. TRIM58 inhibition of GC cell proliferation may be due to β-catenin ubiquitination and inactivation After treatment with shTRIM58 lentivirus and XAV939 in SNU719 cells, (A) Cell proliferation was assessed at 0, 24, 48, and 72 h, and (B) the levels of related proteins (β-catenin, C-myc, Cyclin D1 and survivin) were analyzed by Western blot. (C) A Co-IP shows the co-existence of TRIM58 and β-catenin in the precipitant. (D) β-catenin ubiquitination was examined after infection with TRIM58 lentivirus, and β-catenin expression was detected by western blot. **P < .01 and ***P < .001 compared to shNC + DMSO, ##P < .01 and P < .001 compared to shTRIM58 + DMSO, and ++P < .01 and +++P < .001 compared to shNC + XAV939.
Figure 5. TRIM58 inhibited tumor growth and promoted apoptosis in nude mice through inactivation of β-catenin signaling Twelve nude mice were randomly injected with vector-AGS cells (Six) or TRIM58-AGS cells (Six). (A) The volume and weight of tumor in the mice were measured and a tumor growth curve was drawn. (B) The percentages of apoptosis were examined by Tunel and H&E staining. (C) The protein levels of TRIM58 and β-catenin were determined. (D) β-catenin expression in tissues of nude mice as detected by IHC. *P < .05 and ***P < .001 compared to vector.
