Protein homeostasis is the basis of normal life activities,and the proteasomefamily plays an extremely important function in this process.The proteasome 2os is a concentric circle structure with twoαrings and twoβri...Protein homeostasis is the basis of normal life activities,and the proteasomefamily plays an extremely important function in this process.The proteasome 2os is a concentric circle structure with twoαrings and twoβrings overlapped.The proteasome 2os can perform both ATP-dependent and non-ATP-dependent ubiquitination proteasome degradation by binding to various subunits(such as 19S,11S,and 200 PA),which is performed by its active subunitβ1,β2,andβ5.The proteasome can degrade misfolded,excess proteins tomaintain homeostasis.At the same time,it can be utilized by tumors to degrade over-proliferate and unwanted proteins to support their growth.Proteasomes can affect the development of tumors from several aspects including tumor signaling pathways such as NF-kB and p53,cell cycle,immune regulation,and drug resistance.Proteasome-encoding genes have been found to be overexpressed in a variety of tumors,providing a potential novel target for cancer therapy.In addition,proteasome inhibitors such as bortezomib,carfilzomib,and ixazomib have been put into clinical application as the first-line treatment of multiple myeloma.More and more studies have shown that it also has different therapeutic effects in other tumors such as hepatocellular carcinoma,non-small cell lung cancer,glioblastoma,and neuroblastoma.However,proteasome inhibitors are not much effective due to their tolerance and singleness in other tumors.Therefore,further studies on their mechanisms of action and drug interactions are needed to investigate their therapeutic potential.展开更多
BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on th...BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.展开更多
Per-and polyfluoroalkyl substances(PFASs)can induce a range of adverse health effects,with the precise molecularmechanisms remaining elusive.Extracellular vesicles(EVs)have demonstrated their potential to elucidate un...Per-and polyfluoroalkyl substances(PFASs)can induce a range of adverse health effects,with the precise molecularmechanisms remaining elusive.Extracellular vesicles(EVs)have demonstrated their potential to elucidate unknown molecular mechanisms.Building upon the close alignment of their biological functions with the observed health effects of PFASs,this study innovatively focuses on proteomic insights from EVs into the molecular mechanisms underlying the systemic health effects of PFASs.Through rat exposure experiments and proteomics technology,it not only demonstrated the occurrence of PFASs in EVs but also revealed the alterations in the serum EVs and the expression of their protein cargos following mixed exposure to PFASs,leading to changes in related pathways.These changes encompass various biological processes,including proteasome activity,immune response,cytoskeletal organization,oxidative stress,cell signaling,and nervous system function.Particularly noteworthy is the uncovering of the activation of the proteasome pathway,highlighting significant key contributing proteins.These novel findings provide a new perspective for exploring the molecularmechanism underlying the systemic health effects of PFASs and offer reliable screening for potential biomarkers.Additionally,comparisons with serum confirmed the potential of serum EVs as biological responders and measurable endpoints for evaluating PFASs-induced toxicity.展开更多
The proteasome,an evolutionarily conserved proteolytic complex comprising the 20S core particle and 19S regulatory particles,performs both shared and distinct functions across various tissues and organs.Spermatogenesi...The proteasome,an evolutionarily conserved proteolytic complex comprising the 20S core particle and 19S regulatory particles,performs both shared and distinct functions across various tissues and organs.Spermatogenesis,a highly complex developmental process,relies on proteasome activity at multiple stages to regulate protein turnover.In this study,we selected the 20S subunit PSMA1 and 19S regulatory subunit PSMD2 to investigate the potential functions of the proteasome in spermatogenesis.Using Psma1-EGFP and Psmd2-mCherry knock-in mouse models,we confirmed the expression of both subunits in all spermatogenic cell types,with pronounced presence in early germ cell development.To further clarify their functional significance,we specifically knocked out Psma1 and Psmd2 in germ cells.Deletion of either PSMA1 or PSMD2 led to disrupted spermatogenesis,characterized by the complete absence of sperm in the epididymis.Subsequent analysis indicated that loss of these proteasome components impaired meiotic initiation.Psma1 and Psmd2 knockout germ cells showed accumulation of DMRT1,a key regulator of mitosis-to-meiosis transition,leading to a reduction in STRA8 levels and consequent disruption of meiosis initiation.This study sheds light on the molecular mechanisms that govern meiotic initiation and identifies potential genes associated with male infertility.展开更多
Objectives:Proteasomes,multi-subunit proteases,are key actors of cellular protein catabolism and a number of regulatory processes.The detection of subtle proteasome functioning in tumors may contribute to our understa...Objectives:Proteasomes,multi-subunit proteases,are key actors of cellular protein catabolism and a number of regulatory processes.The detection of subtle proteasome functioning in tumors may contribute to our understanding of the mechanisms of cancer development.The current study aimed to identify the role of low molecular mass protein 2(LMP2),a proteasome immune subunit,in the development of mouse colon 26(C26)adenocarcinoma.Methods:The functions of the LMP2 subunit in tumor development in Balb/c mice were studied using its irreversible inhibitor KZR-504.LMP2 activity was detected by the hydrolysis of the fluorogenic substrate Ac-Pro-Ala-Leu-AMC.Western blotting and Quantitative Reverse Transcription Polymerase Chain Reaction(qRT-PCR)were used.We applied fluorescent tests for cell proliferation and apoptosis.M2 macrophages were obtained by polarization of mouse bone marrow-derived macrophages using the corresponding cytokines.Results:KZR-504 showed high specificity only for the LMP2 subunit and had no negative effect on C26 cells in culture.However,KZR-504 suppressed the formation of tumor conglomerates(by 74%,p<0.001)after C26 cell transplantation in vivo,inhibited the expression of chitinase-<3-like protein 3(Chil3)gene(by 90%,p<0.001),a key marker of immunosuppressive M2 macrophages,in the tumor<microenvironment,and reduced the tumor weight compared to the control(by 48%,p<0.01).KZR-504 also suppressed<the expression of Chil3(by 68%,p<0.05)and arginase-1(Arg1)(by 90%,p<0.001),another marker gene,in M2<<macrophages and violated M0-M2 macrophage polarization in culture.Conclusion:We discovered earlier unknown functions of the proteasome LMP2 subunit to facilitate the formation of tumor conglomerates and maintain Chil3 and Arg1 expression in immunosuppressive M2 macrophages.Our work demonstrates that the proteasome LMP2 subunit can be a target for antitumor treatment.展开更多
Objective To construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotropnic factor （GDNF）, and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by...Objective To construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotropnic factor （GDNF）, and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by transfecting it into bone marrow stromal cells （BMSCs）. Methods pLenti6/V5-GDNF plasmid was set up by double restriction enzyme digestion and ligation, and then the plasmid was transformed into Top10 cells. Purified pLenti6/V5-GDNF plasmids from the positive clones and the packaging mixture were cotransfected to the 293FT packaging cell line by Lipofectamine2000 to produce lentivirus, then the concentrated virus was transduced to BMSCs. Overexpression of GDNF in BMSCs was tested by RT-PCR, ELISA and immunocytochemistry, and its neuroprotection for lactacystin-damaged PC12 cells was evaluated by MTT assay. Results Virus stock of GDNF was harvested with the titer of 5.6×10^5 TU/mL. After tmnsduction, GDNF-BMSCs successfully secreted GDNF to supematant with nigher concentration （800 pg/mL） than BMSCs did （less than 100 pg/mL）. The supematant of GDNF-BMSCs could significantly alleviate the damage of PC12 cells induced by lactacystin （10 μmol/L）. Conclusion Overexpression of lentivirus-mediated GDNF in the BMSCs cells can effectively protect PC12 cells from the injury by the proteasome inhibitor.展开更多
Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycl...Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aimed to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human ovarian cancer SK-OV-3 cells. The results suggested that YSY-01A significantly （P〈0.05） inhibited cellular proliferation of SK-OV-3 cells in a concentration-dependent and time-dependent manner. Furthermore, YSY-01A induced a G2/M cell cycle arrest of SK-OV-3 cells. Further investigation revealed that YSY-01A significantly （P〈0.05） changed the expression levels of a series of cell cycle related protein, such as cyclin B1, cdc2, and p-cdc2 （T14）. Meanwhile, YSY-01A could inhibit the TNF-a-induced NF-kB nuclear translocation and lead to the increase of 1kBa as well as the decrease of IKK and Gadd45a In conclusion, YSY-01A showed remarkable anti-cancer activity on SK-OV-3 cells, and its molecular mechanisms were related to G2/M cell cycle arrest.展开更多
Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycl...Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aims to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human colorectal adenocarcinoma cells HT-29. The results suggested that YSY-01A significantly （P0.05） inhibited cellular proliferation of HT-29 cells in a time and concentration-dependent manner. Furthermore, YSY-01A suppressed the G 2 /M transition of HT-29 cells, whereas the mitotic inhibitor paclitaxel induced M phase accumulation. Further investigation revealed that YSY-01A significantly （P0.05） up-regulated the expression levels of a series of cell cycle related protein, such as cyclin B1, Wee1, p-cdc2 （Tyr15）, p53, p21, and p27. The HT-29 cells only exhibited typical cytotoxic symptom when YSY-01A concentration reached 0.5 μM （P0.05）, which was above the dose we used in the mechanism research. In conclusion, YSY-01A showed remarkable anti-cancer activity on HT-29 cells, and its molecular mechanisms are related to G 2 /M cell cycle transition arrest.展开更多
[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene ...[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).展开更多
As a novel proteasome inhibitor, remarkable proliferation inhibitory effect of compound YSY-01A was shown on tumor cells in previous studies. However, few studies has reported its effect on gastric cancer and related ...As a novel proteasome inhibitor, remarkable proliferation inhibitory effect of compound YSY-01A was shown on tumor cells in previous studies. However, few studies has reported its effect on gastric cancer and related mechanism. We evaluated the anti-proliferative effect of compound YSY-01A using MGC-803 cells and its anti-tumor effect using xenograft nu-BALB/c mouse model. Cell proliferation inhibition was assessed by SRB assay. Related protein expression levels were determined by Western blot assay. We observed that the compound YSY-01A had a significant proliferation inhibitory effect on MGC-803 cells in vitro. Experiment in vivo showed that the compound YSY-01A had a remarkable growth inhibitory effect on MGC-803 cells xenograft tumor when it was used either alone or in combination with the conventional chemotherapeutic agent 5-fluorouracil （5-FU）. Furthermore, YSY-01A and 5-FU had a synergistic effect on xenograft tumor. Results of molecular experiment showed that the compound YSY-01A had a remarkable inhibitory effect on TNF-c~ and IFN induced NF-KB nuclear translocation. At the same time, the compound YSY-01A could reduce the expression of IKK-~, IL-I~ and iNOS, while it significantly enhanced the expression of COX-2 in MGC-803 ceils. Taken together, compound YSY-01A had an impressive tumor inhibitory effect, and it worked in NF-KB-related pathway, suggesting that the compound YSY-01A was an effective therapeutic drug for patients with gastric cancer. Higher tumor cell growth inhibition after the treatment in a combination with 5-FU indicated that combining YSY-01A with 5-FU might be more effective for displaying tumor cell growth inhibitory effects on gastric cancer cells.展开更多
The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of...The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of the two protein complexes in terms of their size and amino acid sequences. Eight groups of subunits from the COP9 signalosome and the proteasome lid complex of different organisms are collected from all the databases at the NCBI website. The corresponding subunits of COP9 signalosome and proteasome lid complex share at least 12% amino acid identity and some conserved regions, and the conserved sites spread evenly over the entire length of the subunits, suggesting that the two complexes have a common evolutionary ancestor. Phylogenetic analyses based on the amino acid sequences of the corresponding subunits of two protein complexes indicate that every tree consists of two clades. The subunits from one of the two protein complexes of different organisms are grouped into one of the two clades respectively. The sequences of single-cell organisms are always the basal groups to that of multi-cell animal and plant species. These results imply that the duplication/divergence events of COP9 signalosome and regulatory lid of the proteasome genes have occurred before the divergence of single-cell and multi-cell eukaryotes, and the genes of the two complexes are independently evolved. The analyses of dN/dS correlation show significant Pearson's correlations between 21 and 15 pairs of subunit-encoding sequences within the COP9 signalosome and the proteasome lid complex respectively, suggesting that those subunits pairs might have related functions and interacted with one another, and resulted in co-evolution.展开更多
On the basis of the Michael-addition mechanism of classical proteasome inhibitors, six dipeptide vinyl sulfonamide and dipeptide vinyl sulfonate derivatives were designed and synthesized. Moreover, an efficient method...On the basis of the Michael-addition mechanism of classical proteasome inhibitors, six dipeptide vinyl sulfonamide and dipeptide vinyl sulfonate derivatives were designed and synthesized. Moreover, an efficient method for the synthesis of g-amino vinyl sulfonamides, key intermediates to the target molecules, was developed via the Wittig-Horner reaction of peptide aldehyde with Wittig reagents derived from methanesulfonamides.展开更多
Five theories shed lights on the potential mechanisms of aging:somatic mutations,telomere loss,mitochondrial defects,and accumulation of altered proteins inside proteasomes.The existence of a program of aging is not y...Five theories shed lights on the potential mechanisms of aging:somatic mutations,telomere loss,mitochondrial defects,and accumulation of altered proteins inside proteasomes.The existence of a program of aging is not yet identified,but overlaps with a program for risks of death.On the other hand,organisms are programmed for survival,which ultimately fails.This failure results in aging,notabily,focusing on alterations of specific genes.Irregular examinations,dysfunctions,insufficient use of fluoride,and removable partial dentures,are favoring the formation of caries and periodontal pathologies.Oral lesions are due to local trauma,related gingival recession,and formation of pockets.They are associated to insufficient removal of food/plaque.Epithelial thinning,and reduction of extracellular matrix components,lead to plications and foldings of the mucosal surface,and subsequently to bacterial colonization.Geriatric dentistry(or gerodontology)is an increasing field of dentistry,mostly associated with the growing percentage of patients over 80+years.展开更多
YSY01-A, as a novel proteasome inhibitor, has shown remarkable proliferation inhibitory effect on certain types of tumor cells. However, few studies have reported its effect on non-small cell lung cancer （NSCLC）, an...YSY01-A, as a novel proteasome inhibitor, has shown remarkable proliferation inhibitory effect on certain types of tumor cells. However, few studies have reported its effect on non-small cell lung cancer （NSCLC）, and its underlying mechanism remains unknown. In our present study, we aimed to figure out the inhibitory effects as well as the mechanism of proteasome inhibitor YSY01-A against A549 cells both individually and in combination with cisplatin. A549 cell proliferation inhibition was assessed by SRB assay. Its related protein expression levels were determined by western blot assay. Moreover, the change of intracellular cisplatin accumulation was examined by ICP-MS assay. The results suggested that YSY01-A significantly （P〈0.001） inhibited the proliferation of A549 cells （IC50 was 36.2 nM for 72 h） in a concentration-dependent and time-dependent manner. Compared with the negative control group, YSY01-A （60 nM, 48 h） down-regulated PI3K/Akt pathway in A549 cells by increasing the expression level of PTEN （P〈0.01）, and decreasing the expression level of PI3K （P〈0.001） and p-Akt/Akt （P〈0.001）. When combined with cisplatin, YSY01-A of different concentrations （5, 10, 20 nM） could significantly increase the inhibition effects on A549 cells compared with the cisplatin alone treatment, showing a synergistic effect. At the same time, YSY01-A could remarkably block the cisplatin-induced down-regulation of hCTR1 in a concentration-dependent manner and increase cisplatin uptake from 2.01 to 2.47 fold （P〈0.001）. In conclusion, compound YSY01-A could significantly inhibit proliferation of NSCLC A549 cells, showing a strong synergistic effect when combined with cisplatin. Down-regulation of PI3K/Akt pathway might be the mechanism of inhibitory effect of YSY0 l-A, and the combination with cisplatin might increase the expression of CTR1 and intracellular cisplatin accumulation.展开更多
There are two degradation systems in mammalian cells, autophagy/lysosomal pathway and ubiquitin-proteasome pathway. Proteasome is consist of multiple protein subunits and plays important roles in degradation of short-...There are two degradation systems in mammalian cells, autophagy/lysosomal pathway and ubiquitin-proteasome pathway. Proteasome is consist of multiple protein subunits and plays important roles in degradation of short-lived cellular proteins. Recent studies reveal that proteasomal degradation system is also involved in signal transduction and regulation of various cellular functions. Dysfunction or dysregulation of proteasomal function may thus be an important pathogenic mechanism in certain neurological disorders. This paper reviews the biological functions of proteasome in signal transduction and its potential roles in neurodegenerative diseases.展开更多
YSY-01A has shown proliferation inhibitory activity to certain types of tumor cells by inhibiting proteasome. How- ever, its effect on autophagy, which is related with the ubiquitin proteasome pathway （UPP）, remains...YSY-01A has shown proliferation inhibitory activity to certain types of tumor cells by inhibiting proteasome. How- ever, its effect on autophagy, which is related with the ubiquitin proteasome pathway （UPP）, remains unclear. Our study aimed to find out its effect on autophagy and possible molecular mechanisms. The results suggested that YSY-0 l A significantly （P〈0.001） inhibited proliferation of PC-3M cells （IC50 was 287 nM for 48 h） in a concentration-dependent and time-dependent manner. YSY-01A （100 nM, 3 h） also induced autophagy in PC-3M ceils through increasing the expression of P53 （P〈0.001）, Beclin-1 （P〈0.001） and LC3 （P〈0.001）, and decreasing the expression of p-roTOR （P〈0.001） as compared with the negative control group. Autophagy stayed at a final stage in PC-3M cells after treated with YSY-01A （400 nM） for 12 h. Meanwhile inhibition of autophagy with chloroquine increased the sensitivity to YSY-01A in PC-3M cells. In conclusion, YSY-01A showed high proliferation inhibitory activity of PC-3M cells and it could induce autophagy in PC-3M cells. Inhibiting autophagy increased the cytotoxic activity of YSY-01A in PC-3M cells.展开更多
基金supported by the National Natural Science Foundation of China(No.82172619)the Natural Science Foundation of Chongqing,China(No.CSTC2021jscx-gksb-No023)the Medical and Industrial Integration Project(China)(No.2022CDJYGRH-002).
文摘Protein homeostasis is the basis of normal life activities,and the proteasomefamily plays an extremely important function in this process.The proteasome 2os is a concentric circle structure with twoαrings and twoβrings overlapped.The proteasome 2os can perform both ATP-dependent and non-ATP-dependent ubiquitination proteasome degradation by binding to various subunits(such as 19S,11S,and 200 PA),which is performed by its active subunitβ1,β2,andβ5.The proteasome can degrade misfolded,excess proteins tomaintain homeostasis.At the same time,it can be utilized by tumors to degrade over-proliferate and unwanted proteins to support their growth.Proteasomes can affect the development of tumors from several aspects including tumor signaling pathways such as NF-kB and p53,cell cycle,immune regulation,and drug resistance.Proteasome-encoding genes have been found to be overexpressed in a variety of tumors,providing a potential novel target for cancer therapy.In addition,proteasome inhibitors such as bortezomib,carfilzomib,and ixazomib have been put into clinical application as the first-line treatment of multiple myeloma.More and more studies have shown that it also has different therapeutic effects in other tumors such as hepatocellular carcinoma,non-small cell lung cancer,glioblastoma,and neuroblastoma.However,proteasome inhibitors are not much effective due to their tolerance and singleness in other tumors.Therefore,further studies on their mechanisms of action and drug interactions are needed to investigate their therapeutic potential.
基金Supported by National Natural Science Foundation of China,No.82160762Guangxi Zhuang Autonomous Region Administration of Traditional Chinese Medicine Scientific Research Project,No.GXZYA20230267+2 种基金China Undergraduate Innovation and Entrepreneurship Training Program,No.S202410598060XChina Undergraduate Innovation and Entrepreneurship Training Program,No.X202410598360Future Academic Star of Guangxi Medical University,No.WLXSZX24074.
文摘BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.
基金supported by the Key Research and Development Plan of Zhejiang Province(No.2021C03176)Zhejiang Provincial Natural Science Foundation of China(No.LY23B070001)+2 种基金the Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province(No.20230009)the National Natural Science Foundation of China(No.22106032)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB0750000).
文摘Per-and polyfluoroalkyl substances(PFASs)can induce a range of adverse health effects,with the precise molecularmechanisms remaining elusive.Extracellular vesicles(EVs)have demonstrated their potential to elucidate unknown molecular mechanisms.Building upon the close alignment of their biological functions with the observed health effects of PFASs,this study innovatively focuses on proteomic insights from EVs into the molecular mechanisms underlying the systemic health effects of PFASs.Through rat exposure experiments and proteomics technology,it not only demonstrated the occurrence of PFASs in EVs but also revealed the alterations in the serum EVs and the expression of their protein cargos following mixed exposure to PFASs,leading to changes in related pathways.These changes encompass various biological processes,including proteasome activity,immune response,cytoskeletal organization,oxidative stress,cell signaling,and nervous system function.Particularly noteworthy is the uncovering of the activation of the proteasome pathway,highlighting significant key contributing proteins.These novel findings provide a new perspective for exploring the molecularmechanism underlying the systemic health effects of PFASs and offer reliable screening for potential biomarkers.Additionally,comparisons with serum confirmed the potential of serum EVs as biological responders and measurable endpoints for evaluating PFASs-induced toxicity.
基金supported by the National Science Fund for Distinguished Young Scholars (81925015)Science and Technology Project of Guangzhou (2023A03J0886,2023A03J0871)National Natural Science Foundation of China (82030039,32400709)。
文摘The proteasome,an evolutionarily conserved proteolytic complex comprising the 20S core particle and 19S regulatory particles,performs both shared and distinct functions across various tissues and organs.Spermatogenesis,a highly complex developmental process,relies on proteasome activity at multiple stages to regulate protein turnover.In this study,we selected the 20S subunit PSMA1 and 19S regulatory subunit PSMD2 to investigate the potential functions of the proteasome in spermatogenesis.Using Psma1-EGFP and Psmd2-mCherry knock-in mouse models,we confirmed the expression of both subunits in all spermatogenic cell types,with pronounced presence in early germ cell development.To further clarify their functional significance,we specifically knocked out Psma1 and Psmd2 in germ cells.Deletion of either PSMA1 or PSMD2 led to disrupted spermatogenesis,characterized by the complete absence of sperm in the epididymis.Subsequent analysis indicated that loss of these proteasome components impaired meiotic initiation.Psma1 and Psmd2 knockout germ cells showed accumulation of DMRT1,a key regulator of mitosis-to-meiosis transition,leading to a reduction in STRA8 levels and consequent disruption of meiosis initiation.This study sheds light on the molecular mechanisms that govern meiotic initiation and identifies potential genes associated with male infertility.
基金funded by the Government program of basic research in Koltzov Institute of Developmental Biology of the Russian Academy of Sciences,number 0088-2024-0009the Ministry of Science and Higher Education of the Russian Federation,project number 075-15-2020-773(NPS).
文摘Objectives:Proteasomes,multi-subunit proteases,are key actors of cellular protein catabolism and a number of regulatory processes.The detection of subtle proteasome functioning in tumors may contribute to our understanding of the mechanisms of cancer development.The current study aimed to identify the role of low molecular mass protein 2(LMP2),a proteasome immune subunit,in the development of mouse colon 26(C26)adenocarcinoma.Methods:The functions of the LMP2 subunit in tumor development in Balb/c mice were studied using its irreversible inhibitor KZR-504.LMP2 activity was detected by the hydrolysis of the fluorogenic substrate Ac-Pro-Ala-Leu-AMC.Western blotting and Quantitative Reverse Transcription Polymerase Chain Reaction(qRT-PCR)were used.We applied fluorescent tests for cell proliferation and apoptosis.M2 macrophages were obtained by polarization of mouse bone marrow-derived macrophages using the corresponding cytokines.Results:KZR-504 showed high specificity only for the LMP2 subunit and had no negative effect on C26 cells in culture.However,KZR-504 suppressed the formation of tumor conglomerates(by 74%,p<0.001)after C26 cell transplantation in vivo,inhibited the expression of chitinase-<3-like protein 3(Chil3)gene(by 90%,p<0.001),a key marker of immunosuppressive M2 macrophages,in the tumor<microenvironment,and reduced the tumor weight compared to the control(by 48%,p<0.01).KZR-504 also suppressed<the expression of Chil3(by 68%,p<0.05)and arginase-1(Arg1)(by 90%,p<0.001),another marker gene,in M2<<macrophages and violated M0-M2 macrophage polarization in culture.Conclusion:We discovered earlier unknown functions of the proteasome LMP2 subunit to facilitate the formation of tumor conglomerates and maintain Chil3 and Arg1 expression in immunosuppressive M2 macrophages.Our work demonstrates that the proteasome LMP2 subunit can be a target for antitumor treatment.
基金This work was supported by the Natural Science Foundation of Shanghai Municipality(No.03ZR14016).
文摘Objective To construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotropnic factor （GDNF）, and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by transfecting it into bone marrow stromal cells （BMSCs）. Methods pLenti6/V5-GDNF plasmid was set up by double restriction enzyme digestion and ligation, and then the plasmid was transformed into Top10 cells. Purified pLenti6/V5-GDNF plasmids from the positive clones and the packaging mixture were cotransfected to the 293FT packaging cell line by Lipofectamine2000 to produce lentivirus, then the concentrated virus was transduced to BMSCs. Overexpression of GDNF in BMSCs was tested by RT-PCR, ELISA and immunocytochemistry, and its neuroprotection for lactacystin-damaged PC12 cells was evaluated by MTT assay. Results Virus stock of GDNF was harvested with the titer of 5.6×10^5 TU/mL. After tmnsduction, GDNF-BMSCs successfully secreted GDNF to supematant with nigher concentration （800 pg/mL） than BMSCs did （less than 100 pg/mL）. The supematant of GDNF-BMSCs could significantly alleviate the damage of PC12 cells induced by lactacystin （10 μmol/L）. Conclusion Overexpression of lentivirus-mediated GDNF in the BMSCs cells can effectively protect PC12 cells from the injury by the proteasome inhibitor.
基金Eleventh Five-Year Plan for National Science and Technology Major Project(Grant No.2009ZX0930-010)National Science Foundation of China(Grant No.81172915)a grant from Major New Drugs Research and Development Platform of Peking University(Grant No.2009ZX09301-010)
文摘Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aimed to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human ovarian cancer SK-OV-3 cells. The results suggested that YSY-01A significantly （P〈0.05） inhibited cellular proliferation of SK-OV-3 cells in a concentration-dependent and time-dependent manner. Furthermore, YSY-01A induced a G2/M cell cycle arrest of SK-OV-3 cells. Further investigation revealed that YSY-01A significantly （P〈0.05） changed the expression levels of a series of cell cycle related protein, such as cyclin B1, cdc2, and p-cdc2 （T14）. Meanwhile, YSY-01A could inhibit the TNF-a-induced NF-kB nuclear translocation and lead to the increase of 1kBa as well as the decrease of IKK and Gadd45a In conclusion, YSY-01A showed remarkable anti-cancer activity on SK-OV-3 cells, and its molecular mechanisms were related to G2/M cell cycle arrest.
基金Eleventh Five-Year Plan for National Science and Technology Major Project (Grant No.2009ZX0930-010)National Science Foundation of China (NSFC81172915)A grant from Major New Drugs Research and Development Platform of Peking University (Grant No.2009ZX09301-010)
文摘Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aims to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human colorectal adenocarcinoma cells HT-29. The results suggested that YSY-01A significantly （P0.05） inhibited cellular proliferation of HT-29 cells in a time and concentration-dependent manner. Furthermore, YSY-01A suppressed the G 2 /M transition of HT-29 cells, whereas the mitotic inhibitor paclitaxel induced M phase accumulation. Further investigation revealed that YSY-01A significantly （P0.05） up-regulated the expression levels of a series of cell cycle related protein, such as cyclin B1, Wee1, p-cdc2 （Tyr15）, p53, p21, and p27. The HT-29 cells only exhibited typical cytotoxic symptom when YSY-01A concentration reached 0.5 μM （P0.05）, which was above the dose we used in the mechanism research. In conclusion, YSY-01A showed remarkable anti-cancer activity on HT-29 cells, and its molecular mechanisms are related to G 2 /M cell cycle transition arrest.
基金Supported by National Basic Research Program of China(2005CB121005)~~
文摘[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).
基金Eleventh Five-Year Plan for National Science and Technology Major Project(Grant No.2009ZX0930-010)National Science Foundation of China(Grant No.81172915)a grant from Major New Drug Research and Development Platform of PekingUniversity(Grant No.2009ZX09301-010)
文摘As a novel proteasome inhibitor, remarkable proliferation inhibitory effect of compound YSY-01A was shown on tumor cells in previous studies. However, few studies has reported its effect on gastric cancer and related mechanism. We evaluated the anti-proliferative effect of compound YSY-01A using MGC-803 cells and its anti-tumor effect using xenograft nu-BALB/c mouse model. Cell proliferation inhibition was assessed by SRB assay. Related protein expression levels were determined by Western blot assay. We observed that the compound YSY-01A had a significant proliferation inhibitory effect on MGC-803 cells in vitro. Experiment in vivo showed that the compound YSY-01A had a remarkable growth inhibitory effect on MGC-803 cells xenograft tumor when it was used either alone or in combination with the conventional chemotherapeutic agent 5-fluorouracil （5-FU）. Furthermore, YSY-01A and 5-FU had a synergistic effect on xenograft tumor. Results of molecular experiment showed that the compound YSY-01A had a remarkable inhibitory effect on TNF-c~ and IFN induced NF-KB nuclear translocation. At the same time, the compound YSY-01A could reduce the expression of IKK-~, IL-I~ and iNOS, while it significantly enhanced the expression of COX-2 in MGC-803 ceils. Taken together, compound YSY-01A had an impressive tumor inhibitory effect, and it worked in NF-KB-related pathway, suggesting that the compound YSY-01A was an effective therapeutic drug for patients with gastric cancer. Higher tumor cell growth inhibition after the treatment in a combination with 5-FU indicated that combining YSY-01A with 5-FU might be more effective for displaying tumor cell growth inhibitory effects on gastric cancer cells.
文摘The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of the two protein complexes in terms of their size and amino acid sequences. Eight groups of subunits from the COP9 signalosome and the proteasome lid complex of different organisms are collected from all the databases at the NCBI website. The corresponding subunits of COP9 signalosome and proteasome lid complex share at least 12% amino acid identity and some conserved regions, and the conserved sites spread evenly over the entire length of the subunits, suggesting that the two complexes have a common evolutionary ancestor. Phylogenetic analyses based on the amino acid sequences of the corresponding subunits of two protein complexes indicate that every tree consists of two clades. The subunits from one of the two protein complexes of different organisms are grouped into one of the two clades respectively. The sequences of single-cell organisms are always the basal groups to that of multi-cell animal and plant species. These results imply that the duplication/divergence events of COP9 signalosome and regulatory lid of the proteasome genes have occurred before the divergence of single-cell and multi-cell eukaryotes, and the genes of the two complexes are independently evolved. The analyses of dN/dS correlation show significant Pearson's correlations between 21 and 15 pairs of subunit-encoding sequences within the COP9 signalosome and the proteasome lid complex respectively, suggesting that those subunits pairs might have related functions and interacted with one another, and resulted in co-evolution.
基金Natural Science Foundation of China for the financial support(Grant No.30772626)
文摘On the basis of the Michael-addition mechanism of classical proteasome inhibitors, six dipeptide vinyl sulfonamide and dipeptide vinyl sulfonate derivatives were designed and synthesized. Moreover, an efficient method for the synthesis of g-amino vinyl sulfonamides, key intermediates to the target molecules, was developed via the Wittig-Horner reaction of peptide aldehyde with Wittig reagents derived from methanesulfonamides.
文摘Five theories shed lights on the potential mechanisms of aging:somatic mutations,telomere loss,mitochondrial defects,and accumulation of altered proteins inside proteasomes.The existence of a program of aging is not yet identified,but overlaps with a program for risks of death.On the other hand,organisms are programmed for survival,which ultimately fails.This failure results in aging,notabily,focusing on alterations of specific genes.Irregular examinations,dysfunctions,insufficient use of fluoride,and removable partial dentures,are favoring the formation of caries and periodontal pathologies.Oral lesions are due to local trauma,related gingival recession,and formation of pockets.They are associated to insufficient removal of food/plaque.Epithelial thinning,and reduction of extracellular matrix components,lead to plications and foldings of the mucosal surface,and subsequently to bacterial colonization.Geriatric dentistry(or gerodontology)is an increasing field of dentistry,mostly associated with the growing percentage of patients over 80+years.
基金Eleventh Five-Year Plan for National Science and Technology Major Project(Grant No.2009ZX0930010)National Science Foundation of China(Grant No.81172915)a grant from Major New Drug Research and Development Platform of Peking University(GrantNo.2009ZX09301010)
文摘YSY01-A, as a novel proteasome inhibitor, has shown remarkable proliferation inhibitory effect on certain types of tumor cells. However, few studies have reported its effect on non-small cell lung cancer （NSCLC）, and its underlying mechanism remains unknown. In our present study, we aimed to figure out the inhibitory effects as well as the mechanism of proteasome inhibitor YSY01-A against A549 cells both individually and in combination with cisplatin. A549 cell proliferation inhibition was assessed by SRB assay. Its related protein expression levels were determined by western blot assay. Moreover, the change of intracellular cisplatin accumulation was examined by ICP-MS assay. The results suggested that YSY01-A significantly （P〈0.001） inhibited the proliferation of A549 cells （IC50 was 36.2 nM for 72 h） in a concentration-dependent and time-dependent manner. Compared with the negative control group, YSY01-A （60 nM, 48 h） down-regulated PI3K/Akt pathway in A549 cells by increasing the expression level of PTEN （P〈0.01）, and decreasing the expression level of PI3K （P〈0.001） and p-Akt/Akt （P〈0.001）. When combined with cisplatin, YSY01-A of different concentrations （5, 10, 20 nM） could significantly increase the inhibition effects on A549 cells compared with the cisplatin alone treatment, showing a synergistic effect. At the same time, YSY01-A could remarkably block the cisplatin-induced down-regulation of hCTR1 in a concentration-dependent manner and increase cisplatin uptake from 2.01 to 2.47 fold （P〈0.001）. In conclusion, compound YSY01-A could significantly inhibit proliferation of NSCLC A549 cells, showing a strong synergistic effect when combined with cisplatin. Down-regulation of PI3K/Akt pathway might be the mechanism of inhibitory effect of YSY0 l-A, and the combination with cisplatin might increase the expression of CTR1 and intracellular cisplatin accumulation.
基金This work was supported by the National Natural Science Foundation of China （No. 30470587, No. 30600197）.
文摘There are two degradation systems in mammalian cells, autophagy/lysosomal pathway and ubiquitin-proteasome pathway. Proteasome is consist of multiple protein subunits and plays important roles in degradation of short-lived cellular proteins. Recent studies reveal that proteasomal degradation system is also involved in signal transduction and regulation of various cellular functions. Dysfunction or dysregulation of proteasomal function may thus be an important pathogenic mechanism in certain neurological disorders. This paper reviews the biological functions of proteasome in signal transduction and its potential roles in neurodegenerative diseases.
基金Eleventh Five-Year Plan for National Science and Technology Major Project(Grant No.2009ZX0930-010)National Science Foundation of China(Grant No.81172915)a grant from Major New Drugs Research and Development Platform of Peking University(Grant No.2009ZX09301-010)
文摘YSY-01A has shown proliferation inhibitory activity to certain types of tumor cells by inhibiting proteasome. How- ever, its effect on autophagy, which is related with the ubiquitin proteasome pathway （UPP）, remains unclear. Our study aimed to find out its effect on autophagy and possible molecular mechanisms. The results suggested that YSY-0 l A significantly （P〈0.001） inhibited proliferation of PC-3M cells （IC50 was 287 nM for 48 h） in a concentration-dependent and time-dependent manner. YSY-01A （100 nM, 3 h） also induced autophagy in PC-3M ceils through increasing the expression of P53 （P〈0.001）, Beclin-1 （P〈0.001） and LC3 （P〈0.001）, and decreasing the expression of p-roTOR （P〈0.001） as compared with the negative control group. Autophagy stayed at a final stage in PC-3M cells after treated with YSY-01A （400 nM） for 12 h. Meanwhile inhibition of autophagy with chloroquine increased the sensitivity to YSY-01A in PC-3M cells. In conclusion, YSY-01A showed high proliferation inhibitory activity of PC-3M cells and it could induce autophagy in PC-3M cells. Inhibiting autophagy increased the cytotoxic activity of YSY-01A in PC-3M cells.
