Severe acute respiratory syndrome coronavirus(SARS-Co V) encodes eight accessory proteins, the functions of which are not yet fully understood. SARS-Co V protein 6(P6) is one of the previously studied accessory protei...Severe acute respiratory syndrome coronavirus(SARS-Co V) encodes eight accessory proteins, the functions of which are not yet fully understood. SARS-Co V protein 6(P6) is one of the previously studied accessory proteins that have been documented to enhance viral replication and suppress host interferon(IFN) signaling pathways. Through yeast two-hybrid screening, we identified eight potential cellular P6-interacting proteins from a human spleen c DNA library. For further investigation, we targeted the IFN signaling pathway-mediating protein, N-Myc(and STAT) interactor(Nmi). Its interaction with P6 was confirmed within cells. The results showed that P6 can promote the ubiquitin-dependent proteosomal degradation of Nmi. This study revealed a new mechanism of SARS-Co V P6 in limiting the IFN signaling to promote SARS-Co V survival in host cells.展开更多
BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by ...BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by inducing apoptosis.However,the mechanism by which matrine regulates antiapoptotic proteins in CRC remains unclear.AIM To identify apoptotic proteins from proteomics and investigate the role of matrine in impeding CRC apoptosis by regulating these proteins.METHODS Tumor and adjacent normal tissues were collected from 52 patients with CRC who underwent surgery between January and December 2021.Data-independent acquisition quantitative proteomic analysis was performed to identify differentially expressed apoptotic proteins.The selected apoptotic proteins were identified through their association with tumor-node-metastasis(TNM)stage and prognosis,then confirmed by immunohistochemical(IHC)staining in validation cohort.In vitro,the role of matrine or apoptotic proteins on cancer cells were analyzed.RESULTS Compared to normal tissues,88 anti-apoptotic proteins from proteomic results were selected.Among them,Shankassociated RH domain interactor(SHARPIN)was identified because of its relationship with TNM stage and overall survival in TCGA database.In the IHC-confirmed cohort,SHARPIN was highly expressed in CRC tissues and localized in the cytoplasm.Higher SHARPIN expression was associated with TNM stage,carbohydrate antigen 153 levels,and gross type compared to low expression.SHARPIN knockdown promoted apoptosis,significantly upregulated the expression of Bcl-2 associated agonist of cell death,Bcl-2 associated X protein,caspase 3,and caspase 8,and downregulated B-cell lymphoma-2(P<0.05).Importantly,matrine treatment promoted apoptosis and reversed the proliferation,invasion,and migration of CRC cells by repressing SHARPIN.CONCLUSION SHARPIN was identified as an upregulated anti-apoptotic protein in CRC,and matrine exhibited anticancer effects by downregulating its expression.Thus,matrine appears to be a promising drug for CRC.展开更多
BACKGROUND Expression of the full-length isoform of Abelson interactor 1(ABI1),ABI1-p65,is increased in colorectal carcinoma(CRC)and is thought to be involved in one or more steps leading to tumor progression or metas...BACKGROUND Expression of the full-length isoform of Abelson interactor 1(ABI1),ABI1-p65,is increased in colorectal carcinoma(CRC)and is thought to be involved in one or more steps leading to tumor progression or metastasis.The ABI1 splice isoform-L(ABI1-SiL)has conserved WAVE2-binding and SH3 domains,lacks the homeodomain homologous region,and is missing the majority of PxxP-and Pro-rich domains found in full-length ABI1-p65.Thus,ABI1-SiL domain structure suggests that the protein may regulate CRC cell morphology,adhesion,migration,and metastasis via interactions with the WAVE2 complex pathway.AIM To investigate the potential role and underlying mechanisms associated with ABI1-SiL-mediated regulation of CRC.METHODS ABI1-SiL mRNA expression in CC tissue and cell lines was measured using both qualitative reverse transcriptase-polymerase chain reaction(RT-PCR)and realtime quantitative RT-PCR.A stably ABI1-SiL overexpressing SW480 cell model was constructed using Lipofectamine 2000,and cells selected with G418.Image J software,CCK8,and transwell assays were used to investigate SW480 cell surface area,proliferation,migration,and invasion.Immunoprecipitation,Western blot,and co-localization assays were performed to explore intermolecular interactions between ABI1-SiL,WAVE2,and ABI1-p65 proteins.RESULTS ABI1-SiL was expressed in normal colon tissue and was significantly decreased in CRC cell lines and tissues.Overexpression of ABI1-SiL in SW480 cells significantly increased the cell surface area and inhibited the adhesive and migration properties of the cells,but did not alter their invasive capacity.Similar to ABI1-p65,ABI1-SiL still binds WAVE2,and the ABI1-p65 isoform in SW480 cells.Furthermore,co-localization assays confirmed these intermolecular interactions.CONCLUSION These results support a model in which ABI1-SiL plays an anti-oncogenic role by competitively binding to WAVE2 and directly interacting with phosphorylated and non-phosphorylated ABI1-p65,functioning as a dominant-negative form of ABI1-p65.展开更多
基金supported by China NSFC grants (#31170152 and 81130083)
文摘Severe acute respiratory syndrome coronavirus(SARS-Co V) encodes eight accessory proteins, the functions of which are not yet fully understood. SARS-Co V protein 6(P6) is one of the previously studied accessory proteins that have been documented to enhance viral replication and suppress host interferon(IFN) signaling pathways. Through yeast two-hybrid screening, we identified eight potential cellular P6-interacting proteins from a human spleen c DNA library. For further investigation, we targeted the IFN signaling pathway-mediating protein, N-Myc(and STAT) interactor(Nmi). Its interaction with P6 was confirmed within cells. The results showed that P6 can promote the ubiquitin-dependent proteosomal degradation of Nmi. This study revealed a new mechanism of SARS-Co V P6 in limiting the IFN signaling to promote SARS-Co V survival in host cells.
基金Supported by National Key Development Plan for Precision Medicine Research,No.2017YFC0910002.
文摘BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by inducing apoptosis.However,the mechanism by which matrine regulates antiapoptotic proteins in CRC remains unclear.AIM To identify apoptotic proteins from proteomics and investigate the role of matrine in impeding CRC apoptosis by regulating these proteins.METHODS Tumor and adjacent normal tissues were collected from 52 patients with CRC who underwent surgery between January and December 2021.Data-independent acquisition quantitative proteomic analysis was performed to identify differentially expressed apoptotic proteins.The selected apoptotic proteins were identified through their association with tumor-node-metastasis(TNM)stage and prognosis,then confirmed by immunohistochemical(IHC)staining in validation cohort.In vitro,the role of matrine or apoptotic proteins on cancer cells were analyzed.RESULTS Compared to normal tissues,88 anti-apoptotic proteins from proteomic results were selected.Among them,Shankassociated RH domain interactor(SHARPIN)was identified because of its relationship with TNM stage and overall survival in TCGA database.In the IHC-confirmed cohort,SHARPIN was highly expressed in CRC tissues and localized in the cytoplasm.Higher SHARPIN expression was associated with TNM stage,carbohydrate antigen 153 levels,and gross type compared to low expression.SHARPIN knockdown promoted apoptosis,significantly upregulated the expression of Bcl-2 associated agonist of cell death,Bcl-2 associated X protein,caspase 3,and caspase 8,and downregulated B-cell lymphoma-2(P<0.05).Importantly,matrine treatment promoted apoptosis and reversed the proliferation,invasion,and migration of CRC cells by repressing SHARPIN.CONCLUSION SHARPIN was identified as an upregulated anti-apoptotic protein in CRC,and matrine exhibited anticancer effects by downregulating its expression.Thus,matrine appears to be a promising drug for CRC.
基金National Natural Science Foundation of China,No.30872923 and No.81672853and Peking University People’s Hospital Scientific Research Development Found,No.RDH2020-11.
文摘BACKGROUND Expression of the full-length isoform of Abelson interactor 1(ABI1),ABI1-p65,is increased in colorectal carcinoma(CRC)and is thought to be involved in one or more steps leading to tumor progression or metastasis.The ABI1 splice isoform-L(ABI1-SiL)has conserved WAVE2-binding and SH3 domains,lacks the homeodomain homologous region,and is missing the majority of PxxP-and Pro-rich domains found in full-length ABI1-p65.Thus,ABI1-SiL domain structure suggests that the protein may regulate CRC cell morphology,adhesion,migration,and metastasis via interactions with the WAVE2 complex pathway.AIM To investigate the potential role and underlying mechanisms associated with ABI1-SiL-mediated regulation of CRC.METHODS ABI1-SiL mRNA expression in CC tissue and cell lines was measured using both qualitative reverse transcriptase-polymerase chain reaction(RT-PCR)and realtime quantitative RT-PCR.A stably ABI1-SiL overexpressing SW480 cell model was constructed using Lipofectamine 2000,and cells selected with G418.Image J software,CCK8,and transwell assays were used to investigate SW480 cell surface area,proliferation,migration,and invasion.Immunoprecipitation,Western blot,and co-localization assays were performed to explore intermolecular interactions between ABI1-SiL,WAVE2,and ABI1-p65 proteins.RESULTS ABI1-SiL was expressed in normal colon tissue and was significantly decreased in CRC cell lines and tissues.Overexpression of ABI1-SiL in SW480 cells significantly increased the cell surface area and inhibited the adhesive and migration properties of the cells,but did not alter their invasive capacity.Similar to ABI1-p65,ABI1-SiL still binds WAVE2,and the ABI1-p65 isoform in SW480 cells.Furthermore,co-localization assays confirmed these intermolecular interactions.CONCLUSION These results support a model in which ABI1-SiL plays an anti-oncogenic role by competitively binding to WAVE2 and directly interacting with phosphorylated and non-phosphorylated ABI1-p65,functioning as a dominant-negative form of ABI1-p65.
