Fig 1: PSMA1 was elevated in gastric cancer.A The real-time PCR assay of the mRNA expression of PSMA1 in gastric cancer tissue and normal tissue is based on the data from the TCGA database. B The basic PSMA1 protein level in different gastric cancer (GC) cell types. C Immunohistochemistry (IHC) staining for PSMA1 in superficial gastritis (SG), intestinal metaplasia (IM), dysplasia (DYS), and GC samples. Bar length: 100 μm. D IHC score analyzed the expression levels of PSMA1 in SG, IM, DYS, and GC. E The Kaplan–Meier plot of overall survival by the expression of PSMA1 in the GC patients, the data was carried out from the TCGA database.
Fig 2: Scheme for the regulatory mechanism of PSMA1 on TAZ.
Fig 3: PSMA1 upregulates TAZ to promote GC cell proliferation in vivo.A Representative image of IHC staining of tumor microarrays from GC patients. Bar length: 200 µm. B IHC scores of PSMA1 and TAZ level between gastric cancer tissue and adjacent normal tissue. C The Kaplan–Meier plot of overall survival by the expression of PSMA1 and TAZ in tumor microarrays related information. D Pearson’s correlation was used to determine the relationship between PSMA1 and TAZ protein expression in gastric cancer tissue specimens. E AGS cells with/without shPSMA1 were injected in nude mice. Representative images of xenograft tumors from mice bearing AGS-shNC and AGS-shPSMA1 cells. F Tumor volume was measured every 3 days. Tumor weight was calculated for each mouse. G PSMA1, YAP, TAZ, and Ki67 expression in the tumor of nude mice were detected by IHC. Bar length: 100 µm. H Western blotting was conducted to measure the level of proliferation-related intratumor protein level.
Fig 4: PSMA1 maintains TAZ stability.A Framework of the TMT labeling quantitative proteomic study. B Upregulated and down-regulated protein by mass spectroscopic analysis. C Volcano plot showing differentially expressed proteins. D Heatmap shows a differentially expressed gene. E Western blot detected YAP and TAZ protein in AGS and BGC-823 cells after transfecting cells with three independent siRNA. Western blot detected YAP and TAZ protein in AGS and BGC-823 cells after transfecting PSMA1-overexpression plasmid. F Western blot detected C-Myc and PCNA protein in AGS and BGC-823 cells after transfecting cells with three independent siRNA or PSMA1-overexpression plasmid.
Fig 5: Oncogenic effect of PSMA1 is dependent on TAZ stabilization.A Colony formation assay was performed to detect the proliferation of GC cells. B Invasion and migration assays were performed to detect the proliferation of AGS cells. Bar length: 100 µm. C Invasion and migration assays were performed to detect the proliferation of BGC-823 cells. Bar length: 100 µm. D CCK-8 assay was used to detect the proliferation of GC cells. E Western blot showing TAZ expression and proteins related to cell growth, such as PCNA and C-Myc in GC cells after a knockdown of PSMA1 and overexpression TAZ or only knockdown PSMA1.
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