Ubiquitin-specific protease 21 promotes tumorigenicity and stemness of colorectal cancer by deubiquitinating and stabilizing ZEB1

BACKGROUND Colorectal cancer(CRC)is one very usual tumor together with higher death rate.Ubiquitin-specific protease 21(USP21)has been confirmed to take part into the regulation of CRC progression through serving as a facilitator.Interestingly,the promotive function of USP21 has also discovered in the progression of CRC.ZEB1 has illustrated to be modulated by USP7,USP22 and USP51 in cancers.However,the regulatory functions of USP21 on ZEB1 in CRC progression need more invest-igations.AIM To investigate the relationship between USP21 and ZEB1 in CRC progression.METHODS The mRNA and protein expressions were assessed through RT-qPCR,western blot and IHC assay.The interaction between USP21 and ZEB1 was evaluated through Co-IP and GST pull down assays.The cell proliferation was detected through colony formation assay.The cell migration and invasion abilities were determined through Transwell assay.The stemness was tested through sphere formation assay.The tumor growth was evaluated through in vivo mice assay.RESULTS In this work,USP21 and ZEB1 exhibited higher expression in CRC,and resulted into poor prognosis.Moreover,the interaction between USP21 and ZEB1 was further investigated.It was demonstrated that USP21 contributed to the stability of ZEB1 through modulating ubiquitination level.In addition,USP21 streng-thened cell proliferation,migration and stemness through regulating ZEB1.At last,through in vivo assays,it was illustrated that USP21/ZEB1 axis aggravated tumor growth.CONCLUSION For the first time,these above findings manifested that USP21 promoted tumorigenicity and stemness of CRC by deubiquitinating and stabilizing ZEB1.This discovery suggested that USP21/ZEB1 axis may provide novel sights for the treatment of CRC.

Ubiquitin-Specific Protease 14 （UBP14） Is Involved in Root Responses to Phosphate Deficiency in Arabidopsis

A mutant isolated from a screen of EMS-mutagenized Arabidopsis lines, per1, showed normal root hair development under control conditions but displayed an inhibited root hair elongation phenotype upon Pi deficiency. Additionally, the per1 mutant exhibited a pleiotropic phenotype under control conditions, resembling Pi-deficient plants in several aspects. Inhibition of root hair elongation upon growth on low Pi media was reverted by treatment with the Pi analog phosphite, suggesting that the mutant phenotype is not caused by a lack of Pi. Reciprocal grafting experiments revealed that the mutant rootstock is sufficient to cause the phenotype. Complementation analyses showed that the PER1 gene encodes an ubiquitin-specific protease, UBP14. The mutation caused a synonymous substitution in the 12th exon of this gene, resulting in a lower abundance of the UBP14 protein, probably as a consequence of reduced translation efficiency. Transcriptional profiling of per1 and wild-type plants subjected to short-term Pi starvation revealed genes that may be important for the signaling of Pi deficiency. We conclude that UBP14 function is crucial for adapting root development to the prevailing local availability of phosphate.

Association of 370-371insACA, 494T〉C, and 1423C〉T haplotype in ubiquitin-specific protease 26 gene and male infertility： a meta-analysis

Whether the 370-371insACA, 494T〉C, and 1423C〉T haplotype in ubiquitin-specific protease 26 （USP26） gene is associated with male infertility is controversial. To clarify this issue, we conducted a meta-analysis based on the most recent studies. Eligible studies were screened by using PubMed and Embase. Pooled odd ratio （OR） with 95% confidence interval （CI） was calculated with fixed effect models. Ten studies with 1603 patients and 2505 controls were included, Overall, the results indicated that there was an association between the haplotype and male infertile risk （OR = 1.74, 95% CI： 1.09-2.77）. The OR calculated based on the five studies in Asia and three in Europe was 1.96 （95% CI： 1,05-3.67） and 1.54 （95% Ch 0.75-3.16） respectively, however, the OR was 0.86 （95% Ch 0.05-15,29） based on the two investigations in America. In addition, the data from the patients with azoospermia （AZO） showed an increased pooled OR of 2.35 （95% Cl： 1.22-4.50）. In contrast, the studies with oligoasthenoteratozoospermia （OAT） exhibited that the pooled OR was 0,97 （95% Ch 0.43-2.16）. Our analyses indicate that there is an association of alteration in USP26 with male infertility, especially in AZO and Asian population.

Novel mutations in ubiquitin-specific protease 26 gene might cause spermatogenesis impairment and male infertility

Aim： To study the incidence of single nucleotide polymorphisms in ubiquitin-specific protease 26 （USP26） gene and its involvement in idiopathic male infertility in China. Methods： Routine semen analysis was performed. Infertility factors such as immunological, infectious and biochemical disorders were examined to select patients with idiopathic infertility. DNA was isolated from peripheral blood of the selected patients and control population, which were examined for mutations using polymerase chain reaction-single strand conformation polymorphism analysis. Furthermore, nucleotide sequences were sequenced in some patients and controls. Results： Of 41 infertile men, 9 （22.0%, P = 0.01） had changes in USP26 gene on the X chromosome. A compound mutation （364insACA; 460G→A） was detected in 8 patients （19.5%, P = 0.01） and a 1044T→A substitution was found in 1 patient （2.4%, P 〉 0.05）. All three variations led to changes in the coding amino acids. Two substitutions predict some changes： 460G→ A changes a valine into an isoleucine, and 1044T → A substitutes a leucine for a phenylalanine. Another insertion of three nucleotides ACA causes an insertion of threonine. No other changes were found in the remaining patients and fertile controls. Conclusion： The USP26 gene might be of importance in male reproduction. Mutations in this gene might be associated with male infertility, and might negatively affect testicular function. Further research on this issue is in progress.

Ubiquitin-specific protease 22 enhances intestinal cell proliferation and tissue regeneration after intestinal ischemia reperfusion injury

BACKGROUND Intestinal ischemia reperfusion(I/R) injury is a serious but common pathophysiological process of many diseases, resulting in a high mortality rate in clinical practice. Ubiquitin-specific protease 22(USP22) acts as regulator of cell cycle progression, proliferation, and tumor invasion. Depleted USP22 expression has been reported to contribute to arrested cell cycle and disrupted generation of differentiated cell types in crypts and villi. However, the role of USP22 in intestinal damage recovery has not been investigated. Therefore, elucidation of the underlying mechanism of USP22 in intestinal I/R injury may help to improve the tissue repair and patient prognosis in clinical practice.AIM To investigate the role of USP22 in intestinal cell proliferation and regeneration after intestinal I/R injury.METHODS An animal model of intestinal I/R injury was generated in male Sprague-Dawley rats by occlusion of the superior mesenteric artery followed by reperfusion.Chiu's scoring system was used to grade the damage to the intestinal mucosa. An in vitro model was developed by incubating rat intestinal epithelial IEC-6 cells in hypoxia/reoxygenation conditions in order to simulate I/R in vivo. siRNA and overexpression plasmid were used to regulate the expression of USP22. USP22,Cyclin D1, and proliferating cell nuclear antigen(PCNA) expression levels were measured by Western blot analysis and immunohistochemistry staining. Cell survival(viability) and cell cycle were evaluated using the Cell Counting Kit-8and flow cytometry, respectively.RESULTS USP22 expression was positively correlated with the expression levels of PCNA and Cyclin D1 both in vivo and in vitro, which confirmed that USP22 was involved in cell proliferation and intestinal regeneration after intestinal I/R injury. Decreased levels of Cyclin D1 and cell cycle arrest were observed in the USP22 knockdown group(P < 0.05), while opposite results were observed in the USP22 overexpression group(P < 0.05). In addition, increased expression of USP22 was related to improved intestinal pathology or IEC-6 cell viability after I/R or hypoxia/reoxygenation. These results suggested that USP22 may exert a protective effect on intestinal I/R injury by regulating cell proliferation and facilitating tissue regeneration.CONCLUSION USP22 is correlated with promoting intestinal cell proliferation and accelerating intestinal tissue regeneration after intestinal I/R injury and may serve as a potential target for therapeutic development for tissue repair during intestinal I/R injury.

Ubiquitin-specific protease 15 contributes to gastric cancer progression by regulating the Wnt/β-catenin signaling pathway

BACKGROUND Ubiquitin-specific protease 15(USP15)is an important member of the ubiquitinspecific protease family,the largest deubiquitinase subfamily,whose expression is dysregulated in many types of cancer.However,the biological function and the underlying mechanisms of USP15 in gastric cancer(GC)progression have not been elucidated.AIM To explore the biological role and underlying mechanisms of USP15 in GC progression.METHODS Bioinformatics databases and western blot analysis were utilized to determine the expression of USP15 in GC.Immunohistochemistry was performed to evaluate the correlation between USP15 expression and clinicopathological characteristics of patients with GC.A loss-and gain-of-function experiment was used to investigate the biological effects of USP15 on GC carcinogenesis.RNA sequencing,immunofluorescence,and western blotting were performed to explore the potential mechanism by which USP15 exerts its oncogenic functions.RESULTS USP15 was up-regulated in GC tissue and cell lines.The expression level of USP15 was positively correlated with clinical characteristics(tumor size,depth of invasion,lymph node involvement,tumor-node-metastasis stage,perineural invasion,and vascular invasion),and was related to poor prognosis.USP15 knockdown significantly inhibited cell proliferation,invasion and epithelialmesenchymal transition(EMT)of GC in vitro,while overexpression of USP15 promoted these processes.Knockdown of USP15 inhibited tumor growth in vivo.Mechanistically,RNA sequencing analysis showed that USP15 regulated the Wnt signaling pathway in GC.Western blotting confirmed that USP15 silencing led to significant down-regulation ofβ-catenin and Wnt/β-catenin downstream genes(c-myc and cyclin D1),while overexpression of USP15 yielded an opposite result and USP15 mutation had no change.Immunofluorescence indicated that USP15 promoted nuclear translocation ofβ-catenin,suggesting activation of the Wnt/β-catenin signaling pathway,which may be the critical mechanism promoting GC progression.Finally,rescue experiments showed that the effect of USP15 on gastric cancer progression was dependent on Wnt/β-catenin pathway.CONCLUSION USP15 promotes cell proliferation,invasion and EMT progression of GC via regulating the Wnt/β-catenin pathway,which suggests that USP15 is a novel potential therapeutic target for GC.

Emerging potential of ubiquitin-specific proteases and ubiquitinspecific proteases inhibitors in breast cancer treatment

Breast cancer is the most frequently diagnosed cancer in women,accounting for 30%of new diagnosing female cancers.Emerging evidence suggests that ubiquitin and ubiquitination played a role in a number of breast cancer etiology and progression processes.As the primary deubiquitinases in the family,ubiquitin-specific peptidases(USPs)are thought to represent potential therapeutic targets.The role of ubiquitin and ubiquitination in breast cancer,as well as the classification and involvement of USPs are discussed in this review,such as USP1,USP4,USP7,USP9X,USP14,USP18,USP20,USP22,USP25,USP37,and USP39.The reported USPs inhibitors investigated in breast cancer were also summarized,along with the signaling pathways involved in the investigation and its study phase.Despite no USP inhibitor has yet been approved for clinical use,the biological efficacy indicated their potential in breast cancer treatment.With the improvements in phenotypic discovery,we will know more about USPs and USPs inhibitors,developing more potent and selective clinical candidates for breast cancer.

Inhibition of Ubiquitin-specific Protease 4 Attenuates Epithelial-Mesenchymal Transition of Renal Tubular Epithelial Cells via Transforming Growth Factor Beta Receptor Type Ⅰ

Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.

USP14抑制对肾缺血再灌注诱导的急性肾损伤及IRE1/JNK信号通路的影响

目的研究USP14的抑制是否可以减轻缺血再灌注(ischemia-reperfusion,IR)诱导的急性肾损伤以及潜在机制。方法通过对小鼠进行左侧肾动脉夹闭45 min和再通建立体内肾IR模型;HE染色观察各组肾组织损伤情况;生化试剂盒检测血尿素氮(blood urea nitrogen,BUN)、肌酐(serum creatinine,Scr)水平;Western blot检测内质网(endoplasmic reticulum,ER)应激相关蛋白(GRP78、CHOP、IRE1和JNK)和凋亡相关蛋白(cleaved capase-3和Bcl-2)的表达;通过TUNEL检测肾组织细胞的凋亡水平。结果与模型组比较,IR组中小鼠肾损伤和肾功能障碍在再灌注期呈时间依赖性逐渐加重,肾组织中USP14的表达和BUN及Scr的水平增加(P<0.05);与IR+sh-NC组比较,IR+sh-USP14组中小鼠肾病理损伤及肾组织中USP14、BUN、Scr的水平降低(P<0.05)。此外,与模型组比较,IR组小鼠肾组织中肾细胞凋亡,cleaved caspase-3和ER应激相关蛋白(GRP78、CHOP、IRE1、p-JNK)的表达升高,Bcl-2的表达降低(P<0.05);与IR+sh-NC组比较,IR+sh-USP14组小鼠肾组织中肾细胞凋亡,cleaved caspase-3和ER应激相关蛋白表达减少,Bcl-2的表达增加(P<0.05)。结论抑制USP14可改善内质网应激和细胞凋亡,降低IRE1/JNK通路蛋白表达,从而减轻肾IR损伤。

Ubiquitin-specific protease 24 promotes EV71 infection by restricting K63-linked polyubiquitination of TBK1

TANK-binding kinase 1(TBK1)is an essential protein kinase for activation of interferon regulatory factor 3(IRF3)and induction of the type I interferons(IFN-I).Although the biochemical regulation of TBK1 activation has been studied,little is known about how enterovirus 71(EV71)employs the deubiquitinases(DUBs)to regulate TBK1 activation for viral immune evasion.Here,we found that EV71 infection upregulated the expression of ubiquitinspecific protease 24(USP24).Further studies revealed that USP24 physically interacted with TBK1,and can reduce K63-linked polyubiquitination of TBK1.Knockdown of USP24 upregulated TBK1 K63-linked polyubiquitination,promoted the phosphorylation and nuclear translocation of IRF3,and in turn improved IFN-I production during EV71 infection.As a consequence,USP24 knockdown dramatically inhibited EV71 infection.This study revealed USP24 as a novel regulator of TBK1 activation,which promotes the understanding of immune evasion mechanisms of EV71 and could provide a potential strategy for treatment of EV71 infection.

去泛素化酶泛素特异性肽酶14促进胰腺癌细胞的增殖和迁移能力

胰腺癌仍然是最致命的癌症之一,而且较少发现有效的治疗方案。虽然在许多肿瘤细胞,包括胰腺癌细胞中观察到泛素特异性蛋白酶14(ubiquitin specific peptidase 14,USP14)的过度表达,但它在胰腺癌中的确切作用仍未得到较好的阐明。我们研究了USP14在胰腺癌中的生物学功能及其分子机制。对癌症基因组图谱数据库(The Cancer Genome Atlas,TCGA)的分析显示USP14在胰腺癌组织中高度表达,进一步探究发现,其表达水平与患者的预后呈负相关。利用shRNAUSP14慢病毒建立了稳定的USP14敲低胰腺癌细胞系,通过CCK8、克隆形成实验、划痕和Transwell试验发现,USP14敲低抑制了胰腺癌细胞的增殖和迁移能力。在胰腺癌细胞SW1990和MIAPaCa2中蛋白质印迹结果表明,下调USP14的表达导致细胞周期蛋白D3(cyclin D3)水平下降,而过表达USP14会增加细胞周期蛋白D3表达水平。此外,免疫共沉淀证明,USP14与细胞周期蛋白D3相互作用,泛素化结果显示,USP14的过表达降低了细胞周期蛋白D3的泛素化水平。并且在SW1990胰腺癌细胞中,CRISPR/Cas9介导的USP14敲除导致细胞周期蛋白D3水平下降。这些发现表明,USP14通过与细胞周期蛋白D3的相互作用促进了胰腺癌细胞的增殖和迁移能力,凸显了USP14作为胰腺癌的潜在治疗靶点。

Circular RNA LPAR3通过miR-143-5p/USP14通路调控食管癌细胞糖酵解的研究

目的探究调控食管癌细胞糖酵解的机制。方法逆转录实时定量PCR(quantitative real-time,qRT-PCR)法分别检测环状RNA溶血磷脂酸受体(circular RNA lysophosphatidic acid receptor 3,CircLPAR3),微小RNA-143-5(microRNA-143-5p,miR-143-5p)和泛素特异性蛋白酶14(ubiquitin-specific protease 14,USP14)的表达水平;使用Seahorse XF 24细胞外通量分析仪测量细胞外酸化率(extracellular acidification rate,ECAR)和耗氧率(oxygen comsumpition rate,OCR);Western印迹法检测糖酵解途径关键酶HK2的表达水平;使用Starbase数据库预测CircLPAR3与miR-143-5p以及miR-143-5p与USP14的靶向结合位点,双荧光素酶试验验证CircLPAR3和miR-143-5p以及miR-143-5p与USP14的靶向关系。结果与正常人食管上皮细胞(normal human esophageal epithelial cells,Het-1A)组比较,食管癌细胞系中CircLPAR3高表达,且KYSE450细胞系的表达最高,同时miR-143-5p低表达而USP14高表达。与Het-1A组比较,KYSE450组HK2的蛋白表达水平增加,同时细胞ECAR增加,整体糖酵解OCR减少;敲低CircLPAR3的表达导致细胞HK2的蛋白表达水平减少,ECAR减少,整体糖酵解OCR增加;在抑制CircLPAR3的表达情况下,抑制miR-143-5p的表达能够部分逆转细胞的糖酵解途径;双荧光素酶实验验证了CircLPAR3和miR-143-5p的靶向关系,以及miR-143-5p和USP14的靶向关系。结论CircLPAR3能够通过调控miR-143-5p/USP14通路调节食管癌细胞糖酵解途径。

黄芪注射液对Balb/c裸鼠银屑病模型的皮损程度及Caspase-14、SOCS1、STAT3水平的影响

目的:研究黄芪注射液对Balb/c裸鼠银屑病模型的皮损程度及Caspase-14、SOCS1、STAT3水平的影响。方法:选取60只Balb/c裸鼠随机数字表分为A、B、C 3组,每组各20只,A组小鼠作为空白对照,B组小鼠作为模型组,C组小鼠作为治疗组。3组小鼠均治疗2周,分析治疗后3组小鼠银屑病PASI评分、小鼠病损皮肤厚度、炎性因子、血清Caspase-14、SOCS1、STAT3水平对比之间的差异。结果:经过治疗后,B组患者的PASI评分显著高于C组,差异具有统计学意义(P<0.05);3组小鼠的治疗后病损皮肤测量比较,差异有统计学意义(P<0.05),通过两两比较,相比空白对照组,B、C两组小鼠的病损皮肤厚度均显著升高,且治疗组小鼠的病损皮肤厚度显著低于模型组,差异具有统计学意义(P<0.05);3组小鼠的治疗后炎性因子之间比较,差异有统计学意义(P<0.05),通过两两比较,相比空白对照组,B、C两组小鼠的炎性因子IL-17、IL-22、 IL-23均显著升高,且治疗组小鼠的炎性因子IL-17、IL-22、 IL-23显著低于模型组,差异有统计学意义(P<0.05);3组小鼠的治疗后血清Caspase-14、SOCS1、STAT3水平比较,差异有统计学意义(P<0.05),通过两两比较,相比空白对照组,B、C两组小鼠的炎性因子血清Caspase-14、SOCS1水平均显著降低,STAT3水平显著升高,且治疗组小鼠的炎性因子aspase-14、SOCS1水平显著低于模型组,STAT3水平显著高于模型组,差异有统计学意义(P<0.05)。结论:黄芪注射液对Balb/c裸鼠银屑病模型的皮损程度有效改善,其可能机制认为是通过对Caspase-14、SOCS1表达水平的降低,降低小鼠的皮损部位的角化程度,改善局部表层增生情况,同时提升STAT3水平,增强局部细胞增殖水平,对于银屑病的康复具有积极意义。

Ubiquitin-specific protease 47 regulates intestinal inflammation through deubiquitination of TRAF6 in epithelial cells

Deubiquitinates(DUBs) alter the stabilities, localizations or activities of substrates by removing their ubiquitin conjugates,which are closely related to the development of inflammatory response. Here, we show that ubiquitin-specific protease 47(USP47) prevents inflammation development in inflammatory bowel disease(IBD). Compared with wild-type mice, Usp47 knockout mice are more susceptible to dextran sodium sulfate(DSS)-induced acute and chronic colitis with higher inflammatory cytokines expression and severe intestinal tissue damage. Chimeric mouse experiments suggest that non-hematopoietic cells mainly contribute to the phenotype. And, DSS-induced colitis of the Usp47 knockout mice depends on commensal bacteria.Mechanistically, down-regulation of USP47 aggravates the activation of NF-κB signaling pathway by increasing the K63-linked poly-ubiquitination of tumor necrosis factor receptor-associated factor 6(TRAF6) in intestinal epithelial cells. Furthermore, the expression of USP47, negatively correlated with the degree of inflammation, is lower at colonic inflammatory lesions than that non-inflammatory sites from the intestine from ulcerative colitis(UC) and Crohn's disease(CD) patients. These data, taken together, indicate that USP47 regulates intestinal inflammation through de-ubiquitination of K63-linked poly-ubiquitination TRAF6 in intestinal epithelial cells.

去泛素化酶USP14调控H_2O_2诱导的H9c2细胞氧化应激损伤

目的:观察抑制泛素特异性蛋白酶14(ubiquitin-specific protease 14,USP14)的活性对H_2O_2诱导的H9c2细胞氧化应激损伤的影响。方法:体外培养H9c2心肌细胞,用H_2O_2(25μmol/L)处理2 h,建立心肌细胞氧化应激损伤模型。将细胞分为对照(control,CON)组、模型组(H_2O_2组)、USP14抑制剂IU1处理组(IU1组)和IU1处理后建模组(IU1+H_2O_2组)。MTS法检测H9c2心肌细胞活力;流式细胞术检测细胞内活性氧簇(ROS)的产生和细胞存活率;Western blot法检测丝裂原活化蛋白激酶家族相关蛋白的表达变化。结果:与CON组相比,H_2O_2组细胞活力、细胞存活率显著降低,细胞内ROS含量、Bax/Bcl-2、P53、p-ERK1/2、p-JNK和p-P38的蛋白水平显著增加(P<0.05);与H_2O_2组比较,IU1+H_2O_2组细胞活力、细胞存活率显著增加,细胞内ROS含量、Bax/Bcl-2、P53、p-ERK1/2、p-JNK和p-P38蛋白水平显著降低(P<0.05)。结论:抑制USP14活性可以减轻氧化应激条件下H9c2心肌细胞的损伤,其机制可能与抑制丝裂原活化蛋白激酶信号活化和下调凋亡相关蛋白有关。

USP14及其抑制剂调控肿瘤的发生与发展

泛素化特异性蛋白酶14(ubiquitin-specific proteases14,USP14)是泛素特异性蛋白酶家族(ubiquitin-specific proteases, USPs)的成员之一。研究发现,USP14可解离靶蛋白质上的泛素标签,通过调控雄性激素受体(androgen receptor, AR)、细胞周期相关蛋白质和凋亡相关蛋白质等多种靶蛋白质的表达水平及活性,在恶性肿瘤的发生发展中发挥重要作用。USP14在多种恶性肿瘤中异常高表达,对肿瘤的调控机制复杂多样,涵盖了肿瘤的基本生命特征,例如细胞增殖、凋亡、炎症反应、自噬及耐药性等。其通过多种信号通路抑制肿瘤细胞凋亡及耐药性;促进炎症反应及肿瘤细胞的增殖活性。同时,在大多数肿瘤中USP14与不良预后密切相关。因此,USP14可作为肿瘤治疗的新型药物靶点,对其抑制剂的开发是抗肿瘤药物研究的新方向。目前,针对USP14的特异性抑制剂主要包括IU1及其类似物IU1-47、IU1-206、IU1-248和b-AP15。因为晶体结构上的差异,IU1的类似物对USP14活性的抑制能力是IU1的10倍。同时研究表明,USP14的抑制剂具有强大的抗肿瘤作用,可通过多种途径来诱导肿瘤细胞死亡,是肿瘤靶向治疗的重要药物。以上研究表明,深入研究USP14及其抑制剂与肿瘤发生发展的相关性极其重要。因此,本文着重对USP14的结构和其在肿瘤中的调控机制进行了综述,总结了USP14抑制剂的研究进展。并进一步探讨USP14相关研究中存在的问题与挑战,以期为未来的研究提供新的方向和策略。

The association between mutations in ubiquitin-specific protease 26(USP26)and male infertility:a systematic review and meta-analysis

During recent decades,the association between mutations in ubiquitin-specific protease 26(USP26)and male infertility remains doubtful.We conducted this meta-analysis to evaluate the association between mutations in USP26 and male infertility according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)2020 guidelines.It was registered in the International Prospective Register of Systematic Reviews(PROSPERO;CRD42021225251).PubMed,Web of Science,and Scopus were systematically searched for comparative clinical studies,which were written in English and provided eligible data.Studies were included when they compared USP26 mutations in azoospermic,oligozoospermic,and asthenozoospermic patients with controls with normal sperm parameter values or whose partners had experienced spontaneous pregnancy.Pooled odds ratio(OR)with 95%confidence interval(CI)was calculated with random effect models.Overall,twelve studies with 3927 infertility patients and 4648 healthy controls were included.The association between overall USP26 mutations and infertility was not significant(OR=1.60,95%CI:0.51-5.01).For specific mutations,the pooled ORs were 1.65(95%CI:1.02-2.69)for cluster mutation(including 370-371insACA,494T>C,and 1423C>T),1.80(95%CI:0.35-9.15)for c.576G>A,1.43(95%CI:0.79-2.56)for c.1090C>T,and 3.59(95%CI:2.30-5.59)for c.1737G>A.Our results suggest that several mutations(cluster mutation,c.1737G>A)may play roles in male infertility,while others(c.576G>A and c.1090C>T)do not show notable associations with male infertility.More high-quality clinical researches are needed for validation.

Deubiquitinase ubiquitin-specific protease 3 (USP3) inhibits HIV-1 replication via promoting APOBEC3G (A3G) expression in both enzyme activity-dependent and -independent manners

Background: Ubiquitination plays an essential role in many biological processes, including viral infection, and can be reversed by deubiquitinating enzymes (DUBs). Although some studies discovered that DUBs inhibit or enhance viral infection by various mechanisms, there is lack of information on the role of DUBs in virus regulation, which needs to be further investigated.Methods: Immunoblotting, real-time polymerase chain reaction,in vivo/in vitro deubiquitination, protein immunoprecipitation, immunofluorescence, and co-localization biological techniques were employed to examine the effect of ubiquitin-specific protease 3 (USP3) on APOBEC3G (A3G) stability and human immunodeficiency virus (HIV) replication. To analyse the relationship between USP3 and HIV disease progression, we recruited 20 HIV-infected patients to detect the levels of USP3 and A3G in peripheral blood and analysed their correlation with CD4^(+) T-cell counts. Correlation was estimated by Pearson correlation coefficients (for parametric data).Results: The results demonstrated that USP3 specifically inhibits HIV-1 replication in an A3G-dependent manner. Further investigation found that USP3 stabilized 90% to 95% of A3G expression by deubiquitinating Vif-mediated polyubiquitination and blocking its degradation in an enzyme-dependent manner. It also enhances the A3G messenger RNA (mRNA) level by binding to A3G mRNA and stabilizing it in an enzyme-independent manner. Moreover, USP3 expression was positively correlated with A3G expression (r= 0.5110) and CD4^(+) T-cell counts (r= 0.5083) in HIV-1-infected patients.Conclusions: USP3 restricts HIV-1 viral infections by increasing the expression of the antiviral factor A3G. Therefore, USP3 may be an important target for drug development and serve as a novel therapeutic strategy against viral infections.

Effects of astragalus injection on the skin lesion degree and Caspase-14,SOCS1 and STAT3 levels of psoriasis model in Balb/c nude mice

Objective: To investigate the effect of Astragalus Injection on the Skin Lesion Degree and Caspase-14, SOCS1 and STAT3 Levels of Psoriasis Model in Balb/c Nude Mice. Methods:Sixty Balb/c nude mice were randomly divided into groups A, B and C with 20 mice in each group. Group A mice were used as blank control, group B mice as model group and group C mice as treatment group. The PASI score of psoriasis, skin thickness, inflammatory factors, serum levels of Caspase-14, SOCS1 and STAT3 in three groups of mice were analyzed after 2 weeks of treatment. Result: After treatment, the P ASI score of group B was significantly higher than that of group C, with statistical significance (P < 0.05);there was statistical significance in the measurements of lesion skin of three groups of mice after treatment (P <0.05). Compared with the blank control group, the thickness of lesion skin in group B and C was significantly higher, and the thickness of lesion skin in treatment group was significantly lower than that in control group (P < 0.05). Compared with the blank control group, the inflammatory factors IL-17, IL-22 and IL-23 in the B and C groups were significantly increased, and the inflammatory factors IL-17, IL-22 and IL-23 in the treatment group were significantly lower than those in the model group. The levels of serum C aspase-14, SOCS1 and STAT3 in three groups of mice were significantly different after treatment (P < 0.05). Compared with the blank control group, the levels of serum C aspase-14 and SOCS1 in B and C groups were significantly lower and the levels of STAT3 were significantly higher, and the levels of inflammatory factors aspase-14 and SO in treatment group were significantly higher than those in control group. The level of CS1 was significantly lower than that of model group, and the level of STAT3 was significantly higher than that of model group (P <0.05). Conclusion: Astragalus membranaceus injection can effectively improve the degree of psoriasis in Balb/c nude mice. Its possible mechanism is that it can decrease the expression of Caspase-14 and SOCS1, reduce the degree of keratosis in the lesion site of mice, improve the local surface hyperplasia, increase the level of STAT3 and enhance the level of local cell proliferation, which is of positive significance for the rehabilitation of psoriasis.

去泛素化酶USP14对铁死亡的调控作用

铁死亡是一种依赖铁的新型非凋亡细胞死亡形式,在调节细胞代谢、细胞命运决定以及疾病的发生和发展中扮演着关键角色。去泛素化酶14(ubiquitin-specific protease 14,USP14)作为泛素-蛋白酶体和自噬途径的重要调节因子,除了参与细胞周期、免疫反应、信号转导等经典生命活动的调节,对铁死亡相关途径的关键蛋白发挥了精细调控作用。USP14作为一个潜在的抗肿瘤药物靶点,目前已经有一系列靶向抑制剂被开发出来。本文主要综述了USP14的活性调控机制、在铁死亡相关疾病中的生理作用以及其小分子抑制剂的研究进展,并对相关研究中的问题和挑战进行了深入探讨,旨在为未来的研究提供新的方向和策略。