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
Background Integrase interactor 1 （INI1）,which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex,has been identified as a tumor suppressor in many tumors.Nonetheless,the role of INI1 in ...Background Integrase interactor 1 （INI1）,which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex,has been identified as a tumor suppressor in many tumors.Nonetheless,the role of INI1 in gastric tumor progression is not known exactly.The aim of this research was to investigate the effect of INI1 in the carcinogenesis and progression of gastric cancer.Methods Gastric tumor tissues with different differentiation levels from clinical gastric carcinoma samples and adjacent control normal tissues were taken. Expression levels of INI1 were detected by quantitative reverse transcriptation-polymerase chain reaction （RT-PCR） and Western blotting.Gastric cancer cell line SGC7901 was transfected with INI1 eukaryotic expressing vector INI1-GFP.Cell proliferation activities were assessed by MTT; cell count and cell cycle were detected by flow cytometry （FCM）; cell apoptosis were measured by TUNEL and FCM; cell migration and invasiveness were evaluated by wound healing and transwell assays.Expression levels of INI1 and proliferation-related genes including p16,p21,cyclin D1 and cyclin A,apoptosis genes p53,B-cell non-Hodgkin lymphoma-2 （Bc/-2）,Bcl-2-associated x protein （Bax） and caspase-3,and invasion-related genes including intercellular adhesion molecule 1 （ICAM1）,matrix metalloproteinase 2 （MMP2）,MMP9 and tissue inhibitor of matrix metalloproteinase 1 （T/MP1）,were detected by quantitative RT-PCR and Western blotting.Results INI1 expression levels were lower in gastric carcinoma compared with adjacent control normal tissues.Overexpression of INI1 in SGC7901 cells inhibited its proliferation and invasiveness,but increased anoikis and Go/G1 cell number.INI1-GFP transfection upregulated expression of INI1 and proliferation related genes p16 and p21,apoptosis genes p53 and Bax,and invasion-related genes TIMP1; cyclin D1,cyclin A,Bcl2,ICAM1,MMP2 and MMP9 were downregulated,and there was no significant change in caspase 3 levels.Conclusion INI1 plays a key role in gastric carcinogenesis by affecting proliferation,apoptosis and invasion.展开更多
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
Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflu...Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflux and AUXIN RESISTANT 1(AUX1)for auxin influx.However,how the auxin gradient is established remains elusive.Here we identified a new mutant with a short root,strong auxin distribution in the lateral root cap and an impaired gravitropic response.The causal gene encoded an Arabidopsis homolog of the human unconventional prefoldin RPB5 interactor(URI).At URI interacted with prefoldin 2(PFD2)and PFD6,twoβ-type PFD members that modulate actin and tubulin patterning in roots.The auxin reporter DR5_(rev):GFP showed that asymmetric auxin redistribution after gravistimulation is disordered in aturi-1 root tips.Treatment with the endomembrane protein trafficking inhibitor brefeldin A indicated that recycling of the auxin transporter PIN2 is disrupted in aturi-1 roots as well as in pfd mutants.We propose that At URI cooperates with PFDs to recycle PIN2and modulate auxin distribution.展开更多
Interactors introduced in another recent paper by the authors are fourth order tensors which describes the interacting effects on the effective elastic properties of damaged solids due to spatial distribution of defe...Interactors introduced in another recent paper by the authors are fourth order tensors which describes the interacting effects on the effective elastic properties of damaged solids due to spatial distribution of defects. Based on the concept of interactors, an interacting model for the effective elastic properties of anisotropically damaged solids is proposed. As an application of the proposed model, the anisotropic effective elastic properties of an isotropic matrix material with circular holes in periodically rectangular distribution are theoretically estimated up to third order terms of the porosity and the estimation is compared with numerical results.展开更多
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.
基金This work was supported by grants from the National Natural Science Foundation of China （No.81072033）,Hebei Provincial Natural Science Foundation （No. C2010000619）, Extra Characteristic Foundation of Colleges and Universities in Hebei Province （No.[2005]52）,Health Department of Hebei Province （No.20100119）.
文摘Background Integrase interactor 1 （INI1）,which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex,has been identified as a tumor suppressor in many tumors.Nonetheless,the role of INI1 in gastric tumor progression is not known exactly.The aim of this research was to investigate the effect of INI1 in the carcinogenesis and progression of gastric cancer.Methods Gastric tumor tissues with different differentiation levels from clinical gastric carcinoma samples and adjacent control normal tissues were taken. Expression levels of INI1 were detected by quantitative reverse transcriptation-polymerase chain reaction （RT-PCR） and Western blotting.Gastric cancer cell line SGC7901 was transfected with INI1 eukaryotic expressing vector INI1-GFP.Cell proliferation activities were assessed by MTT; cell count and cell cycle were detected by flow cytometry （FCM）; cell apoptosis were measured by TUNEL and FCM; cell migration and invasiveness were evaluated by wound healing and transwell assays.Expression levels of INI1 and proliferation-related genes including p16,p21,cyclin D1 and cyclin A,apoptosis genes p53,B-cell non-Hodgkin lymphoma-2 （Bc/-2）,Bcl-2-associated x protein （Bax） and caspase-3,and invasion-related genes including intercellular adhesion molecule 1 （ICAM1）,matrix metalloproteinase 2 （MMP2）,MMP9 and tissue inhibitor of matrix metalloproteinase 1 （T/MP1）,were detected by quantitative RT-PCR and Western blotting.Results INI1 expression levels were lower in gastric carcinoma compared with adjacent control normal tissues.Overexpression of INI1 in SGC7901 cells inhibited its proliferation and invasiveness,but increased anoikis and Go/G1 cell number.INI1-GFP transfection upregulated expression of INI1 and proliferation related genes p16 and p21,apoptosis genes p53 and Bax,and invasion-related genes TIMP1; cyclin D1,cyclin A,Bcl2,ICAM1,MMP2 and MMP9 were downregulated,and there was no significant change in caspase 3 levels.Conclusion INI1 plays a key role in gastric carcinogenesis by affecting proliferation,apoptosis and invasion.
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 an initiation fund from the Shandong Agricultural University,the National Natural Science Foundation of China(31570291)the Shandong“Foreign experts double hundred”Program(WST2017008)the Natural Science Foundation of Shandong Province(ZR2021MC175)。
文摘Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflux and AUXIN RESISTANT 1(AUX1)for auxin influx.However,how the auxin gradient is established remains elusive.Here we identified a new mutant with a short root,strong auxin distribution in the lateral root cap and an impaired gravitropic response.The causal gene encoded an Arabidopsis homolog of the human unconventional prefoldin RPB5 interactor(URI).At URI interacted with prefoldin 2(PFD2)and PFD6,twoβ-type PFD members that modulate actin and tubulin patterning in roots.The auxin reporter DR5_(rev):GFP showed that asymmetric auxin redistribution after gravistimulation is disordered in aturi-1 root tips.Treatment with the endomembrane protein trafficking inhibitor brefeldin A indicated that recycling of the auxin transporter PIN2 is disrupted in aturi-1 roots as well as in pfd mutants.We propose that At URI cooperates with PFDs to recycle PIN2and modulate auxin distribution.
文摘Interactors introduced in another recent paper by the authors are fourth order tensors which describes the interacting effects on the effective elastic properties of damaged solids due to spatial distribution of defects. Based on the concept of interactors, an interacting model for the effective elastic properties of anisotropically damaged solids is proposed. As an application of the proposed model, the anisotropic effective elastic properties of an isotropic matrix material with circular holes in periodically rectangular distribution are theoretically estimated up to third order terms of the porosity and the estimation is compared with numerical results.
