ABSTRACT
Pancreatic cancer remains one of the deadliest cancer types with few effective treatment options. While the overexpression of ubiquitin-specific protease 14 (USP14) has been observed in many tumor cells, including pancreatic cancer cells, its precise role in pancreatic cancer is not well defined. Here, we investigated the biological function of USP14 in pancreatic cancer and its molecular mechanisms. Our analysis of the Cancer Genome Atlas database revealed that USP14 was highly expressed in pancreatic cancer tissues,and further investigation revealed that its expression level was negatively correlated with the prognosis of patients. In SW1990 and MIAPaCa2 pancreatic cancer cells,we established stable USP14-knockdown cell lines using the shRNA-USP14 lentivirus and found that USP14 knockdown inhibited the proliferation and migration ability of pancreatic cancer cells by CCK8, colony formation assay, wound-healing and Transwell assays. Western blotting analysis showed that downregulation of USP14 expression resulted in a decrease in CyclinD3 protein levels, while overexpression of USP14 increased the protein levels in SW1990 and MIAPaCa2 pancreatic cancer cells. Furthermore, co-immunoprecipitation demonstrated that USP14 interacts with CyclinD3 and ubiquitination assays show that overexpression of USP14 reduces the ubiquitination level of CyclinD3. Moreover, CRISPR / Cas9-mediated USP14 knockout in SW1990 pancreatic cancer cells resulted in decreased CyclinD3 protein levels. These findings suggest that USP14 promotes the proliferation and migration ability of pancreatic cancer cells by interacting with CyclinD3, highlighting USP14 as a potential therapeutic target for pancreatic cancer.
ABSTRACT
Hazardous environmental factors as well as occupational factors can lead to elevated incidence of diseases including tumors, and specific molecular biomarkers are needed to guide the diagnosis and treatment of diseases. In recent years, ubiquitin-specific protease 14 (USP14) has gradually attracted the attention of researchers. USP14 is widely expressed in various organs of human body and regulates the stability and degradation of important proteins in various signaling pathways. Studies have shown that its abnormal expression is highly correlated with tumors, neurodegenerative diseases, autophagy, immune response, and viral infections, and is involved in the regulation of various classic signaling pathways. It has been shown to play a key role in the development of various human diseases and can be used as a diagnostic and prognostic molecular biomarker and therapeutic target in the development of tumors. This paper reviewed the current status of research on the structure and regulation of USP14 and its function in physiological and pathological processes, with the aim of providing a reference for research on diseases or injuries caused by environmental and occupational factors.
ABSTRACT
Ubiquitin-specific protease 14 (USP14), a member of the ubiquitin-specific proteases family, plays an important role in the development and progression of malignant tumors through removing ubiquitin chain from substrates such as androgen receptor (AR), cell cycle-related proteins and apoptosis-related proteins to regulate protein stability and activity. The mechanisms of USP14 in a variety of malignant tumors are complex and diverse, including promotion in cell proliferation and inflammation as well as inhibition in apoptosis, autophagy, and drug resistance. Simultaneously, aberrant expression of USP14 is closely correlated with poor prognosis in most malignant tumors. Therefore, USP14 is a potential drug target for tumor therapy, and the development of its inhibitors appears as a new direction of anti-tumor drug research. Currently, specific inhibitors of USP14 mainly include IU1, its analogs (IU1-47, IU1-206, IU1-248), and b-AP15. The inhibitory ability of IU1 analogs on USP14 activity is 10 times than that of IU1, due to the difference in crystal structure. On the other hand, the inhibitors of USP14 show powerful anti-tumor effects inducing tumor cell death through a variety of signal pathways, which enables the inhibitor as potential targeted drugs. It will be extremely important to further explore the relationship between USP14 and its inhibitors in the tumor development and progression. This review summarizes the latest research of USP14 and its inhibitors, including their structures and functions in malignant tumors. Furthermore, the problems, challenges, new directions and strategies for future research, were also reviewed in current study.
ABSTRACT
Polycomb group (PcG) proteins are transcriptional repressors that control cell fate and the development of embryo, and they elicit function mainly in the form of polycomb repressive complex (PRC). Chromobox protein homolog 6 (CBX6) is one of the core protein subunits of PRC1, which plays an important role in gene expression regulation, cell renewal and differentiation, tumorigenesis and development, and stem cell maintenance. In this study, CBX6 was found to be degraded through a ubiquitin-proteasome dependent pathway. Then the gene expression library containing 92 deubiquitinating enzymes (DUB) was used to screen DUB targeting CBX6 and results found that ubiquitin-specific protease 29 (USP29) could obviously stabilize CBX6 protein level and extend its half-life (P < 0.05). Immunoprecipitation experiments found that CBX6 interacted with USP29 through its C-terminal domains; Further studies found that USP29 regulated the protein stability of CBX6 by deubiquitination in an enzymatic-activity dependent manner. Cell proliferation assay also found that USP29 inhibited the proliferation of MCF7 cells (P<0.0001). Taken together, through screening, this study found that USP29 could stabilize CBX6 protein level through deubiquitinating CBX6 and inhibit the cell proliferation of MCF7.
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@# 【Objective】To investigate the inhibitory effect and mechanism of microRNA-30a-5p（miR-30a-5p）on epithelial mesenchymal transition in cervical cancer Hela cells.【Methods】Hela cervical cancer cell lines were transfected with miR-30a-5p mimics or negative control mimic，respectively，as 30a-5p or NC group. Control group was established with untreated Hela cervical cancer cells. miR-30a-5p content in each group was detected by RT-PCR assay. Transwell assay was used to detect the invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of N- cadherin，α-Catenin and ubiquitin specific processing peptidase 22（USP22）protein in the 3 groups. Prediction target genes of miR-30a-5p by bioinformatics methods. Antagonistic effect of USP22 over-expression on miR-30a-5p inhibi⁃tion of EMT was detected by western blot assay. The relationship between miR-30a-5p and USP22 was detected by dual luciferase assay. Subcutaneous transplantation tumor model established，and the effect of miR-30a-5p in vivo was ob⁃ served.【Results】The miR-30a-5p intracellular quantity in 30a-5p group Hela cells was up-regulated，and the expres⁃ sion level of miR-30a-5p was 853.82（862.26~843.11）times higher than that of Control group（P＜0.01）. The number of invasive cells in 30a-5p group was 8.17（8.32~8.03），which was significantly lower than that of Control group 62.33 （63.52~60.19）（P＜0.01）. USP22 may be the target gene of miR-30a-5p. In 30a-5p group，the intracellular quantity of N-cadherin protein was decreased，the intracellular quantity of α-Catenin protein was increased，and the intracellular quantity of USP22 protein was decreased. The intracellular quantity of N-cadherin protein in 30a-5p group was down-reg⁃ ulated，the intracellular quantity of α-Catenin protein was increased，and the intracellular quantity of USP22 protein was reduced. After USP22 over-expression，the intracellular quantity of N-cadherin protein in cervical cancer cells of 30a-5p group was up-regulated，and the intracellular quantity of α-Catenin protein were down-regulated. Dual luciferase assay showed that USP22 is a downstream target gene of miR-30a-5p（P＜0.01）. Subcutaneous transplantation tumors in 30a- 5p group were significantly smaller than those in Control group.【Conclusion】miR-30a-5p may inhibit the expression of EMT related protein through the downstream target gene USP22，and the epithelial mesenchymal transition function of cervical cancer Hela cells.
ABSTRACT
Chromosomal instability, leading to aneuploidy, is one of the hallmarks of human cancers. USP44 (ubiquitin specific peptidase 44) is an important molecule that plays a regulatory role in the mitotic checkpoint and USP44 loss causes chromosome mis-segregation, aneuploidy and tumorigenesis in vivo. In this study, it was investigated the immunoexpression of USP44 in 28 malignant salivary gland neoplasms and associated the results with DNA ploidy status assessed by image cytometry. USP44 protein was widely expressed in most of the tumor samples and no clear association could be established between its expression and DNA ploidy status or tumor size. On this basis, it may be concluded that the aneuploidy of the salivary gland cancers included in this study was not driven by loss of USP44 protein expression.
Resumo Instabilidade cromossômica acarretando aneuploidia é um dos fatores marcantes de neoplasias malignas humanas. USP44 (peptidase específica de ubiquitina 44) é uma importante molécula que exerce um papel regulador no ciclo celular e sua perda pode acarretar em segregação cromossômica deficiente, aneuploidia e desenvolvimento de tumores in vivo. Neste estudo, investigou-se a expressão imuno-histoquímica da proteína USP44 em 28 neoplasias malignas de glândulas salivares, associando-se os resultados com o estado de ploidia do DNA avaliado por citometria de fluxo. A proteína USP44 apresentou ampla expressão na maioria das amostras avaliadas e não foi observada associação entre a expressão protéica e o estado de ploidia do DNA ou extensão do tumor. Baseando-se nos resultados, concluiu-se que a aneuploidia das neoplasias malignas de glândulas de salivares incluídas neste estudo não foi influenciada pela perda de expressão da proteína USP44.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Aneuploidy , DNA/genetics , Salivary Gland Neoplasms/genetics , Ubiquitin-Specific Proteases/metabolismABSTRACT
AIM: To evaluate the effect of inhibiting ubiquitin-specific protease 14 (USPl4) activity on oxidative stress induced by H2O2 of H9c2 cells.METHODS: The H9c2 cells were incubated with H2O2 at 25 μmol/L for 2 h to establish the oxidative stress injury model.The cells were divided into control group, H2O2 group, IU1 group (25 μmol/L or 50 μmol/L) and IU1+ H2O2 group.The H9c2 cells activity was measured by MTS assay.The level of intracellular reactive oxygen species (ROS) and cell survival rate were analyzed by flow cytometry assay.The changes of the mitogen-activated protein kinase (MAPK) family related proteins were detected by Western blot.RESULTS: Compared with control group, the cell activity and the viability rate in H2O2 group were decreased (P<0.05), while the intracellular ROS, the protein levels of Bax/Bcl-2, P53, p-ERK1/2, p-JNK and p-P38 were increased (P<0.05).Compared with H2O2 group, the cell activity and the viability rate of the H9c2 cells in IU1+H2O2 group were increased (P<0.05), while the intracellular ROS, the protein levels of Bax/Bcl-2, P53, p-ERK1/2, p-JNK and p-P38 were decreased (P<0.05).CONCLUSION: Inhibition of USPl4 activity reduces the oxidative stress injury of the H9c2 cells.The mechanism may be related to inhibition of the MAPK signaling and down-regulation of apoptosis related proteins.
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Objective@#To investigate the expression characteristics of the USP24 gene in the mouse testis and its role in spermatogenesis.@*METHODS@#We examined the expression characteristics of USP24 in the testis tissues of wild-type mice at different postnatal weeks (PNW) and androgen receptor (AR)-knockout (ARKO) adult mice using real-time quantitative PCR and immunofluorescence, and detected the transcriptional activity of the USP24 promoter by dual-luciferase reporter gene assay.@*RESULTS@#The expression of the USP24 gene was low in the testis tissue of the wild-type mice at PNW 1, increased dramatically at PNW 3 and stayed at a similar level till PNW 8. The USP24 protein was located mainly in the cytoplasm of Sertoli and spermatogenic cells. Compared with the wild-type, the adult ARKO mice showed a decreased expression of USP24 localized in the posterior head and mid-piece of the mature sperm in the testis. Dual-luciferase reporter gene assay showed that the transcriptional activity of the USP24 promoter was increased after testosterone stimulation.@*CONCLUSIONS@#The increased expression of the USP24 gene was associated with the initiation of sexual development, and the USP24 protein was expressed in the mature sperm of the mice. USP24 is an AR-target gene, which may be involved in the regulation of spermatogenesis in mice.
Subject(s)
Animals , Male , Mice , Mice, Knockout , Promoter Regions, Genetic , Receptors, Androgen , Genetics , Sertoli Cells , Spermatogenesis , Genetics , Spermatozoa , Metabolism , Testis , Metabolism , Testosterone , Transcription, Genetic , Ubiquitin Thiolesterase , Genetics , MetabolismABSTRACT
Objective This study is to investigate the influences of USP7 on the cytobiological characteristics of laryngeal cancer cells by small interfering RNA (siRNA) interfering the USP7 expression in the laryngeal cancer cells .Methods The self‐de‐signed highly efficient siRNA was used to conduct the specific interference on USP7 expression in laryngeal cancer HEP2 cells . Then the influence on the capacity of cell proliferation and migration ,as well as apoptosise after USP7 interference were observed by using the CCK‐8 method ,Transwell chamber migration test and flow cytometry .Results The self‐designed siRNA could effi‐ciently inhibit the expression of USP7 mRNA in laryngeal cancer cells ,furthermore markedly suppressed the proliferation and mi‐gration of laryngeal cancer cells ,enhanced the cell apoptosis in laryngeal cancer HEP2 cells in vitro .Conclusion The siRNA inter‐fering USP7 can inhibit the proliferation and migration capacity of laryngeal cancer cells ,and promoted their apoptosis .
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Objective There is close relationship between ubiquitin-specific protease 9X(USP9X) and the biological behav-ior of some tumor.The aim of this study is to investigate the expression and clinical significance of USP 9X in pancreatic carcinoma of elderly patients. Methods The expression of USP9X was detected in 30 pieces of surgically resected primary pancreatic carcinoma tissue and adjacent nontumorous pancreatic tissue of elderly patients by streptavidin -perosidase immunohistochemical method . Results The rate of USP9X positive expression was 56.7%, there was not positive expression in adjacent nontumorous pancreatic tissue .There was no relation between the expression of USP 9X with gender, age, the tumor positin, the tumor size and degree of differentiation (P>0.05), while it was significantly correlated with lymph node metastasis and TNM stages (P<0.05).By using Cox proportional haz-ards analysis, multivariable analysis revealed that TNM stages , lymph node metastasis and USP9X expression were independent risk factor(P<0.05). Conclusion The results indicated that USP9X may play a role in the pathogenesis and prognosis of pancreatic cancer of elderly patients .
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The ubiquitin-specific protease ( USP) inhibitors influence many crucial cellular activities and some immune processes, such as anti-inflammatory, anti-infection and anti-tumor by silencing the functions of USP.The main USP inhibitors, which potency and specificity are underlined and current methods for detecting and identifying USP inhibitors are discussed of in this review.
ABSTRACT
Ubiquitin specific peptidases play important roles in controlling proteasome activity,organogenesis,tumorigenesis and transcriptional regulation.Ubiquitin specific peptidase 9X is able to remove ubiquitin from substrate proteins,so it may effectively regulate the proliferation,adhesion and signal transduction of tumor cells.It is a one of the most important factors in tumor progression.