BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CC...BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.展开更多
The heavy biofouling on electrochemical sensor surface poses a formidable challenge for biosensing in human blood.Herein,we designed a multilayer filtering-sensing sandwich patch that served as a versatile platform to...The heavy biofouling on electrochemical sensor surface poses a formidable challenge for biosensing in human blood.Herein,we designed a multilayer filtering-sensing sandwich patch that served as a versatile platform to surmount the substantial fouling constraints for detection in human blood.The patch integrated two functional layers:(i) Inspired by dialysis phenomenon,a filtering-mass transfer hydrophilic membrane with heterogeneous nanostructure was used to filter large-size substances(like cells,bacteria and microorganisms,etc.) and continuously pass through the rest of the biological fluid(like proteins,metabolites and inorganic salts,etc.).(ii) the polypeptide composite hydrogel(r GO/PEPG) on the screenprinted electrode(SPE) surface,with the modulation of-COOH and-NH_2 groups,endowed a strong hydrophilic layer with electric neutrality to further facilitate the antifouling ability.Notably,the integration of the filtering porous membrane with the antifouling hydrogel ensures the strong antifouling ability of the electrochemical sensor in complex human blood.Furthermore,the self-healing property of the r GO/PEPG,relying on the physical π-π stacking forces,aligns the electrochemical sensor with practical needs.The constructed antifouling biosensor based on the filtering-sensing sandwich patch was successfully applied for the sensitive detection of cortisol in human blood,with an acceptable accuracy comparable to the enzyme-linked immunosorbent assay(ELISA) method.The strategy presented herein represent a promising advance along the road to construct effective antifouling biosensing devices with robust operation in diverse complex body fluids.展开更多
基金Supported by Shandong Provincial Natural Science Foundation,No.ZR2023MH329Project of Shandong Province Higher Educational Youth Innovation Science and Technology Program,No.2023KJ263and Natural Science Foundation of Gansu Province,China,No.22JR5RA953.
文摘BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.
基金supported by the National Natural Science Foundation of China (Nos.22174082,22374085)the Key Research and Development Program of Shandong Province (No.2021ZDSYS30)Natural Science Foundation of Shandong Province,China (No.ZR2024QB059)。
文摘The heavy biofouling on electrochemical sensor surface poses a formidable challenge for biosensing in human blood.Herein,we designed a multilayer filtering-sensing sandwich patch that served as a versatile platform to surmount the substantial fouling constraints for detection in human blood.The patch integrated two functional layers:(i) Inspired by dialysis phenomenon,a filtering-mass transfer hydrophilic membrane with heterogeneous nanostructure was used to filter large-size substances(like cells,bacteria and microorganisms,etc.) and continuously pass through the rest of the biological fluid(like proteins,metabolites and inorganic salts,etc.).(ii) the polypeptide composite hydrogel(r GO/PEPG) on the screenprinted electrode(SPE) surface,with the modulation of-COOH and-NH_2 groups,endowed a strong hydrophilic layer with electric neutrality to further facilitate the antifouling ability.Notably,the integration of the filtering porous membrane with the antifouling hydrogel ensures the strong antifouling ability of the electrochemical sensor in complex human blood.Furthermore,the self-healing property of the r GO/PEPG,relying on the physical π-π stacking forces,aligns the electrochemical sensor with practical needs.The constructed antifouling biosensor based on the filtering-sensing sandwich patch was successfully applied for the sensitive detection of cortisol in human blood,with an acceptable accuracy comparable to the enzyme-linked immunosorbent assay(ELISA) method.The strategy presented herein represent a promising advance along the road to construct effective antifouling biosensing devices with robust operation in diverse complex body fluids.
