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Knockdown of USP32 inhibits cell growth. (A) The mRNA level of USP32 was reduced by siUSP32-2261 and siUSP32-386. Data are represented as mean ± SD of three biological replicates. (B) Left panel: Representative images of three independent western bolting analyses showing the knockdown efficiency of siUSP32-2261 and siUSP32-386. Right panel: statistical quantification of left panel. (C) MTT assay determining the cell viability of U-87 MG and U-251 MG. Data are represented as mean ± SD of six biological replicates. (D) Representative images of three independent colony formation assays showing that USP32 knockdown inhibited cell proliferation. (E) Statistical quantification of (D). (F) USP32 knockdown reduced the number of EdU⁺ cells. Bar: 10 μm. (G) Statistical quantification of (F). Data are represented as mean ± SD of three technical replicates. (H) Representative images of three independent cell cycle assays by flow cytometry. (I) Histogram showing the percentage of each cell-cycle phase in (H). One way ANOVA followed by Tukey’s post-hoc test: vs siRNA NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
Source publication
Glioblastoma (GBM) patients present poor prognosis. Deubiquitination by deubiquitinating enzymes (DUBs) is a critical process in cancer progression. Ubiquitin-specific proteases (USPs) constitute the largest sub-family of DUBs. Evaluate the role of USP32 in GBM progression and provide a potential target for GBM treatment. Clinical significance of U...
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Hepatocellular carcinoma (HCC) is one of the deadliest cancers. The retinoblastoma protein (RB1), a regulator of cell proliferation, is functionally inactivated in HCC by CYCLIN D/E-mediated phosphorylation. However, the mechanism of RB1-inactivation is unclear because only small percentages of HCCs exhibit amplification of CYCLIN D/E or mutations...
Citations
... Aberrant expression of USP32 triggers certain diseases such as Parkinson's, Fragile X Syndrome, Chronic Kidney Disease and Cancer [11,[17][18][19]. Recent research has demonstrated the high expression of USP32 in a range of cancers [20][21][22][23][24][25][26] and its role in the initiation and progression of cancer. ...
... For example, USP32 in uences the invasion, migration, and proliferation of small cell lung cancer [21]; USP32 affects the development of epithelial ovarian cancer [20], glioblastoma [22], and gastric cancer [23] through the regulation of related proteins. USP32 interacts with Rab35 due to the deubiquitinating enzyme activity of USP32, therefore promoting the mesenchymal malignancies in the gastrointestinal tract to acquire imatinib resistance [27]. ...
The regulatory significance of ubiquitin-specific peptidase 32 (USP32) in tumor is significant, nevertheless, the biological roles and regulatory mechanisms of USP32 in non-small cell lung cancer (NSCLC) remain unclear. According to our research, USP32 was strongly expressed in NSCLC cell lines and tissues and was linked to a bad prognosis for NSCLC patients. Interference with USP32 resulted in a significant inhibition of NSCLC cell proliferation, migration potential, and EMT development; on the other hand, USP32 overexpression had the opposite effect. To further elucidate the mechanism of action of USP32 in NSCLC, we screened H1299 cells for interacting proteins and found that USP32 interacts with BAG3 (Bcl2-associated athanogene 3) and deubiquitinates and stabilizes BAG3 in a deubiquitinating activity-dependent manner. Functionally, restoration of BAG3 expression abrogated the antitumor effects of USP32 silencing. Furthermore, USP32 increased the phosphorylation level of the RAF/MEK/ERK signaling pathway in NSCLC cells by stabilizing BAG3. In summary, these findings imply that USP32 is critical to the development of NSCLC and could offer a theoretical framework for the clinical diagnosis and management of NSCLC patients in the future.
... In 2021, USP32 was found to be overexpressed in epithelial ovarian cancer (EOC), particularly in metastatic peritoneal tumors, and it positively regulates the proliferation and epithelial-mesenchymal transition (EMT) capabilities of cancer cells [9]. In 2022 and 2023, researchers confirmed that USP32 act as an oncogene in glioblastoma, gastrointestinal stromal tumors (GISTs) and acute myeloid leukemia through performing in vivo and in vitro experiments [10][11][12]. Through a series of preliminary pan-cancer analyses, we discovered that USP32 is significantly overexpressed in several cancer types such as BC, cholangiocarcinoma, esophageal carcinoma, head and neck squamous cell carcinoma, HCC and GC (Fig. 1A). ...
Background
Ubiquitin-specific protease 32 (USP32) is a highly conserved gene that promotes cancer progression. However, its role in hepatocellular carcinoma (HCC) is not well understood. The aim of this project is to explore the clinical significance and functions of USP32 in HCC.
Methods
The expression of USP32 in HCC was evaluated using data from TCGA, GEO, TISCH, tissue microarray, and human HCC samples from our hospital. Survival analysis, PPI analysis and GSEA analysis were performed to evaluate USP32-related clinical significance, key molecules and enrichment pathways. Using the ssGSEA algorithm and TIMER, we investigated the relationships between USP32 and immune infiltrates in the TME. Univariate and multivariate Cox regression analyses were then used to identify key USP32-related immunomodulators and constructed a USP32-related immune prognostic model. Finally, CCK8, transwell and colony formation assays of HCC cells were performed and an HCC nude mouse model was established to verify the oncogenic role of USP32.
Results
USP32 is overexpressed in HCC and its expression is an independent predictive factor for outcomes of HCC patients. USP32 is associated with pathways related to cell behaviors and cancer signaling, and its expression is significantly correlated with the infiltration of immune cells in the TME. We also successfully constructed a USP32-related immune prognostic model using 5 genes. Wet experiments confirmed that knockdown of USP32 could repress the proliferation, colony formation and migration of HCC cells in vitro and inhibit tumor growth in vivo.
Conclusion
USP32 is highly expressed in HCC and closely correlates with the TME of HCC. It is a potential target for improving the efficacy of chemotherapy and developing new strategies for targeted therapy and immunotherapy in HCC.
... We queried USP32 in human tissues through the Human Protein Atlas website, the testis showed the greatest RNA expression of USP32 (Fig. 3). USP32 is upregulated in a variety of cancers, including small cell lung cancer [24], gastric cancer [25,26], breast cancer [23,27,28], epithelial ovarian cancer [29], glioblastoma [30], gastrointestinal stromal tumor [31], pancreatic duct adenocarcinoma [32] and acute myeloid leukemia [33]. Endogenous USP32 is found in the cytoplasm and membrane, according to the findings of a subcellular separation experiment [34] and a fluorescence protection experiment [23]. ...
... The elevated expression of USP32 protein in GBM tissues was validated by a study. Glioblastoma cell lines (Umur118 MG, Umur87 MG, A172, T98G, and Umur251 MG) had higher mRNA and protein levels of USP32 than normal brain cells SVG p12 [30]. In addition, they found that the higher the expression of USP32 in GBM, the worse the prognosis. ...
... Cancer cell proliferation and migration can be stopped in vitro by inhibiting USP32, and tumor growth can be stopped in vivo by down-regulating the USP32 gene. In their research, they discovered that USP32 can speed up the transition of the cell cycle from the G0 phase to the G1 phase [30], start DNA replication, and so enhance the growth of cancer cells. Then, RT-qPCR tests were used to confirm the effects of USP32 on a few functional molecules. ...
An essential protein regulatory system in cells is the ubiquitin-proteasome pathway. The substrate is modified by the ubiquitin ligase system (E1-E2-E3) in this pathway, which is a dynamic protein bidirectional modification regulation system. Deubiquitinating enzymes (DUBs) are tasked with specifically hydrolyzing ubiquitin molecules from ubiquitin-linked proteins or precursor proteins and inversely regulating protein degradation, which in turn affects protein function. The ubiquitin-specific peptidase 32 (USP32) protein level is associated with cell cycle progression, proliferation, migration, invasion, and other cellular biological processes. It is an important member of the ubiquitin-specific protease family. It is thought that USP32, a unique enzyme that controls the ubiquitin process, is closely linked to the onset and progression of many cancers, including small cell lung cancer, gastric cancer, breast cancer, epithelial ovarian cancer, glioblastoma, gastrointestinal stromal tumor, acute myeloid leukemia, and pancreatic adenocarcinoma. In this review, we focus on the multiple mechanisms of USP32 in various tumor types and show that USP32 controls the stability of many distinct proteins. Therefore, USP32 is a key and promising therapeutic target for tumor therapy, which could provide important new insights and avenues for antitumor drug development. The therapeutic importance of USP32 in cancer treatment remains to be further proven. In conclusion, there are many options for the future direction of USP32 research.
... USP32 belongs to the ubiquitin-specific protease family, deubiquitinating enzymes which have been reported to be involved in several cancer initiation and progression (ovarian cancer [34], gastric cancer [35], glioblastoma [36], breast cancer [37], small cell lung cancer [38]). A recent work suggests a pivotal role of USP32 in pancreatic ductal adenocarcinoma given its higher expression levels compared to normal pancreatic tissues and a significant association with tumor grade and stage [39]. ...
... However, these data must be considered preliminary and need to be confirmed on a larger sample of patients. The two other patients who were followed-up after medical treatment without statistically significant USP32 Ovarian cancer Overexpressed [34] Gastric cancer Overexpressed [35] Glioblastoma Overexpressed [36] Breast cancer Overexpressed [37] Small cell lung cancer Overexpressed [38] Pancreatic ductal adenocarcinoma ...
Background
Neuroendocrine tumors (NETs) early diagnosis is a clinical challenge that require a deep understanding of molecular and genetic features of this heterogeneous group of neoplasms. However, few biomarkers exist to aid diagnosis and to predict prognosis and treatment response. In the oncological field, tumor-educated platelets (TEPs) have been implicated as central players in the systemic and local responses to tumor growth, thereby altering tumor specific RNA profile. Although TEPs have been found to be enriched in RNAs, few studies have investigated the potential of a type of RNA, circular RNAs (circRNA), as platelet-derived biomarkers for cancer. In this proof-of-concept study, we aim to demonstrate whether the circRNAs signature of tumor educated platelets can be used as a liquid biopsy biomarker for the detection of gastroenteropancreatic (GEP)-NETs and the prediction of the early response to treatment.
Methods
We performed a 24-months, prospective proof-of-concept study in men and women with histologically proven well-differentiated G1-G2 GEP-NET, aged 18–80 years, naïve to treatment. We performed a RNAseq analysis of circRNAs obtained from TEPs samples of 10 GEP-NETs patients at baseline and after 3 months from therapy (somatostatin analogs or surgery) and from 5 patients affected by non-malignant endocrinological diseases enrolled as a control group.
Results
Statistical analysis based on p < 0.05 resulted in the identification of 252 circRNAs differentially expressed between GEP-NET and controls of which 109 were up-regulated and 143 were down-regulated in NET patients. Further analysis based on an FDR value ≤ 0.05 resulted in the selection of 5 circRNAs all highly significant downregulated. The same analysis on GEP-NETs at baseline and after therapy in 5 patients revealed an average of 4983 remarkably differentially expressed circRNAs between follow-up and baseline samples of which 2648 up-regulated and 2334 down-regulated, respectively. Applying p ≤ 0.05 and FDR ≤ 0.05 filters, only 3/5 comparisons gave statistically significant results.
Conclusions
Our findings identified for the first time a circRNAs signature from TEPs as potential diagnostic and predictive biomarkers for GEP-NETs.
... However, an open question is how RAB7 monoubiquitination contributes to USP32 depletion-dependent defects in EGFR degradation and tumorigenesis. Notably, our and others' work has already linked USP32 to breast and other cancers, highlighting the implications of deregulated USP32 function [19][20][21]. ...
Behind the scenes of signaling cascades initiated by activated receptors, endocytosis determines the fate of internalized proteins through degradation in lysosomes or recycling. Over the years, significant progress has been made in understanding the mechanisms of endocytosis and deregulation in disease states. Here we review the role of the EGF-SNX3-EGFR axis in breast cancers with an extended discussion on deregulated EGFR endocytosis in cancer.
... Ubiquitin-specific protease 32 (USP32) is recognized as a new member of the ubiquitin-specific proteases subfamily. It alters protein stability and localization, thereby regulating their activity in different pathological stages of many human diseases, like cancer (6)(7)(8). So far, it has been reported that USP32 was over-expressed in lung and breast cancers, enhancing cellular proliferation and tissue metastasis (9). ...
Objective:
Gastric cancer is the fifth most common neoplasm and the fourth reason for mortality globally. Incidence rates are highly variable and dependent on risk factors, epidemiologic and carcinogenesis patterns. Previous studies reported that Helicobacter pylori (H. pylori) infection is one the strongest known risk factor for gastric cancer. USP32 is a deubiquitinating enzyme identified as a potential factor associated with tumor progression and a key player in cancer development. On the other hand, SHMT2 is involved in serine-glycine metabolism to support cancer cell proliferation. Both USP32 and SHMT2 are reported to be upregulated in many cancer types, including gastric cancer, but its complete mechanism is not fully explored yet. The present study explored possible mechanism of action of USP32 and SHMT2 in the progression of gastric cancer.
Materials and methods:
In this experimental study, Capsaicin (0.3 g/kg/day) and H. pylori infection combination was used to successfully initiate gastric cancer conditions in mice. It was followed by 40 and 70 days of treatment to establish initial and advanced conditions of gastric cancer.
Results:
Histopathology confirmed formation of signet ring cell and initiation of cellular proliferation in the initial gastric cancer. More proliferative cells were also observed. In addition, tissue hardening was confirmed in the advanced stage of gastric cancer. USP32 and SHMT2 showed progressive upregulated expression, as gastric cancer progress. Immunohistologically, it showed signals in abnormal cells and high-intensity signals in the advanced stage of cancer. In USP32 silenced tissue, expression of SHMT2 was completely blocked and reverted cancer development as evident with less abnormal cell in initial gastric cancer. Reduction of SHMT2 level to one-fourth was observed in the advanced gastric cancer stages of USP32 silenced tissue.
Conclusion:
USP32 had a direct role in regulating SHMT2 expression, which attracted therapeutic target for future treatment.
... USP32 was reported to be one protein that contributed to the chimeric Tre2 (USP6) oncogene [154]. The pro-cancer effects of USP32 have been observed in breast cancer and glioblastoma [155,156]. Additionally, USP32 promotes GC cell growth, metastasis, and chemoresistance by upregulating SMAD2, an important protein in the TGF-β signaling pathway [55]. However, it is unclear whether USP32 regulates the ubiquitination level of SMAD2. ...
Gastric cancers (GCs) are malignant tumors with a high incidence that threaten global public health. Despite advances in GC diagnosis and treatment, the prognosis remains poor. Therefore, the mechanisms underlying GC progression need to be identified to develop prognostic biomarkers and therapeutic targets. Ubiquitination, a post-translational modification that regulates the stability, activity, localization, and interactions of target proteins, can be reversed by deubiquitinases (DUBs), which can remove ubiquitin monomers or polymers from modified proteins. The dysfunction of DUBs has been closely linked to tumorigenesis in various cancer types, and targeting certain DUBs may provide a potential option for cancer therapy. Multiple DUBs have been demonstrated to function as oncogenes or tumor suppressors in GC. In this review, we summarize the DUBs involved in GC and their associated upstream regulation and downstream mechanisms and present the benefits of targeting DUBs for GC treatment, which could provide new insights for GC diagnosis and therapy.
