Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana....Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana. Whether TCP1 can directly bind to the promoter region of DWF4, however, has not been experimentally demonstrated. Here we provide our biochemical and genetic evidence that TCP1 mediates the expression of DWF4 by directly associating with the two GGNCCC motifs in the promoter region of DWF4. The expression levels of DWF4 are positively correlated to TCP1 abundance in planta. Electrophoretic mobility shift assays （EMSAs） using various synthetic DNA fragments suggest that the GGNCCC core sequence is critical for TCP1 binding. DNA sequences flanking the GGNCCC motifs are less important for the association of TCPI. Using DWF4p-GUS transgenic plants as an assay system, it is clearly indicated that these motifs are required for the positive regulation of DWF4 transcription by TCP1. More significantly, whole genome microarray analyses indicate that TCP1 can directly or indirectly regulate the expression of many other genes known to be important for normal plant growth and development.展开更多
目的:探讨沉默TCP1α蛋白对人胰腺导管腺癌细胞的影响。方法:采用SiRNA干扰技术沉默两种人胰腺癌细胞(PANC-1和Mia Pa Ca-2)中TCP1α蛋白基因,Western blot检测3种TCP1α蛋白(TCP1α-1、TCP1α-2、TCP1α-3)表达情况,验证SiRNA沉默效果...目的:探讨沉默TCP1α蛋白对人胰腺导管腺癌细胞的影响。方法:采用SiRNA干扰技术沉默两种人胰腺癌细胞(PANC-1和Mia Pa Ca-2)中TCP1α蛋白基因,Western blot检测3种TCP1α蛋白(TCP1α-1、TCP1α-2、TCP1α-3)表达情况,验证SiRNA沉默效果。以体外增殖实验(MTT实验)和Transwell迁移侵袭实验验证抑制TCP1α蛋白表达对两种胰腺癌细胞体外增殖、迁移及侵袭等生物学行为的影响。Western blot检测PD98059和CHX共同处理的基因沉默人胰腺癌细胞的TCPIα蛋白表达情况。结果:沉默TCP1α基因后两种人胰腺癌细胞增殖和迁移侵袭能力显著降低。结论:抑制TCP1α蛋白表达能明显降低人胰腺导管腺癌细胞的增殖侵袭及迁移能力,且能诱发人胰腺导管腺癌细胞的急性凋亡。展开更多
AIM:To investigate the value of chaperonin containing TCP1,subunit 3(CCT3) to predict the prognosis of patients with hepatocellular carcinoma(HCC) and determine its function in HCC progression.METHODS:CCT3 expression ...AIM:To investigate the value of chaperonin containing TCP1,subunit 3(CCT3) to predict the prognosis of patients with hepatocellular carcinoma(HCC) and determine its function in HCC progression.METHODS:CCT3 expression levels were examined in human non-cancerous liver tissues and a variety of HCC cell lines by quantitative real-time PCR and immunoblotting.CCT3 expression was suppressed by small interfering RNA.The effects of reducing CCT3 expression in HCC cells were tested.The3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide(MTT) assay,cell counting experiment,cell cycle assay,apoptosis assay and invasion assay were employed to evaluate cell functions in vitro.Immunohistochemistry was performed on HCC specimens.In addition,CCT3 expression in HCC specimens was also assessed at the protein and mRNA level.Associations between clinicopathological characteristics and prognosis were analyzed,along with the possible mechanisms involved in CCT3's function in HCC progression.RESULTS:The expression levels of CCT3 mRNA and protein were upregulated in HCC cell lines in contrast to adjacent non-cancerous tissues.Reducing CCT3 expression not only suppressed cell proliferation in cell counts,MTT assay,cell cycle assay and induced cell apoptosis(P < 0.05 vs negative control),but also inhibited the tumor cell invasion capacity in vitro {P< 0.01 vs negative control).Overexpression of CCT3 in the nuclei of cancer cells in HCC specimens(58of 104 patients,55.8%) was associated with poor prognosis in HCC patients(3-year survival rate,55.5%vs 84.2%,P = 0.020) after hepatectomy.Mechanistic analyses showed that signal transducer and activator of transcription 3(STAT3) activation was decreased even when stimulated by interleukin-6 after knocking down CCT3 in the HepG2 cell line.CONCLUSION:Overexpression of CCT3 in the nuclei of cancerous cells is associated with HCC progression.CCT3 may be a target that affects the activation of STAT3 in HCC.展开更多
TCP transcription factors are essential for plant growth and environmental response,but little is known about their specific roles in growth,hormone response,and stress response in tea plants.Here,CsTCP5 and CsTCP18 w...TCP transcription factors are essential for plant growth and environmental response,but little is known about their specific roles in growth,hormone response,and stress response in tea plants.Here,CsTCP5 and CsTCP18 were cloned from the tea variety'Longjing 43',and subcellular localization revealed that both CsTCP5 and CsTCP18 proteins were present in the nucleus.Exogenous abscisic acid(ABA)and methyl jasmonate(MeJA)had different effects on CsTCP5 and CsTCP18 expression,but both genes were repressed by drought and low-temperature treatment.Yeast-two hybrid(Y2H)and bimolecular fluorescence complementation(BiFC)assays demonstrated that neither transcription factor was transcriptionally active but that each interacted with members of the CsMYB and CsJAZ families.Transgenic Arabidopsis plants overexpressing CsTCP5 had smaller cotyledons and fewer lateral roots compared with wild-type(WT)and empty vector(EV)Arabidopsis,and their root growth and germination rate were inhibited by ABA and MeJA treatment.Lateral root numbers also decreased significantly in CsTCP18-OE Arabidopsis after MeJA treatment.These results demonstrate that CsTCP5 and CsTCP18 have regulatory effects on cotyledon development,lateral root growth,and germination rate,and these effects can be modulated by ABA and MeJA.Under drought stress,the CsTCP5-OE and CsTCP18-OE lines exhibited greater survival,lower plant height,smaller rosette leaves,delayed flowering,increased activities of antioxidant enzymes,decreased MDA content,and increased proline content compared with WT and EV Arabidopsis.These findings suggest that CsTCP5 and CsTCP18 are important for tea plant growth,interact with ABA and MeJA pathways,and play roles in stress response.展开更多
(Molecular Plant8(3):482–485;March 2015;https://doi.org/10.1016/j.molp.2014.11.018)The supplemental figure S3(panel A)was amended since the picture of the yeast control spot TCP14-AD/M5RGL2BD(-W-L medium)has been use...(Molecular Plant8(3):482–485;March 2015;https://doi.org/10.1016/j.molp.2014.11.018)The supplemental figure S3(panel A)was amended since the picture of the yeast control spot TCP14-AD/M5RGL2BD(-W-L medium)has been used in place of TCP15-AD/M5GAI-BD.The authors apologize for not detecting this error prior to publication and for any inconvenience that may have caused.展开更多
Dear Editor,Molecular chaperones are critical mediators of oncogenesis and necessary for cell survival.1 The cytoplasmic chaperone TRiC(Tcomplex protein-1 ring complex,also known as CCT)comprises two back-to-back stac...Dear Editor,Molecular chaperones are critical mediators of oncogenesis and necessary for cell survival.1 The cytoplasmic chaperone TRiC(Tcomplex protein-1 ring complex,also known as CCT)comprises two back-to-back stacked rings,with each ring containing eight subunits(CCT1–CCT8).2 And the effects of CCT subunits on tumors may be different.However,the roles of the CCT1 subunit(also known as TCP1)in hepatocellular carcinoma(HCC)and its molecular mechanism have not been investigated thoroughly.Understanding these details can provide new ideas and strategies for treating HCC.展开更多
BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 s...BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 subunit 3(CCT3)expression and its clinical implications,and its effects on LSCC cell growth.METHODS Systematic data on CCT3 mRNA expression were collected from biomedical databases,and integrated further based on the standardized mean difference and the summary receiver operating characteristic curve.Single-cell RNA-seq data were mined to validate the expression level of CCT3 mRNA.In-house immunohistochemistry was performed to explore the CCT3 protein levels of clinical LSCC samples and their relationship with clinical parameters.The growth function of LSCC cell was analyzed using CRISPR knockout screening.CCT3-related signaling pathway analyses were conducted using gene set enrichment analysis.Protein-protein interaction network construction was performed to identify hub genes.RESULTS CCT3 mRNA was significantly overexpressed in 269 LSCC tissues cases across multiple independent datasets(standardized mean difference=32,area under the curve=0.93);At the translational level,the in-house immunohistochemical analysis further demonstrated the consistent upregulation of CCT3 protein in 88 cases of LSCC samples(58 non-LSCC samples vs 30 LSCC samples,P=1.4e^(-14)).Analysis of clinical parameters showed no significant differences among subgroup.Functional characterization with clustered regularly interspaced short palindromic repeats--mediated gene knockout revealed that depletion of CCT3 potently suppressed LSCC cell viability in vitro.Gene set enrichment analysis indicated that CCT3 was markedly associated with several key oncogenic pathways,including extracellular matrix receptor interaction and cell cycle regulation pathways.CONCLUSION CCT3 upregulation in LSCC may influence cellular growth by regulating related pathways,indicating its potential as a biomarker and therapeutic target for LSCC.展开更多
基金supported by the National Basic Research Program of China(Grant No.2011CB915401)the National Natural Science Foundation of China(Grant Nos. 90917019,91117008 and 91317311)
文摘Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana. Whether TCP1 can directly bind to the promoter region of DWF4, however, has not been experimentally demonstrated. Here we provide our biochemical and genetic evidence that TCP1 mediates the expression of DWF4 by directly associating with the two GGNCCC motifs in the promoter region of DWF4. The expression levels of DWF4 are positively correlated to TCP1 abundance in planta. Electrophoretic mobility shift assays （EMSAs） using various synthetic DNA fragments suggest that the GGNCCC core sequence is critical for TCP1 binding. DNA sequences flanking the GGNCCC motifs are less important for the association of TCPI. Using DWF4p-GUS transgenic plants as an assay system, it is clearly indicated that these motifs are required for the positive regulation of DWF4 transcription by TCP1. More significantly, whole genome microarray analyses indicate that TCP1 can directly or indirectly regulate the expression of many other genes known to be important for normal plant growth and development.
文摘目的:探讨沉默TCP1α蛋白对人胰腺导管腺癌细胞的影响。方法:采用SiRNA干扰技术沉默两种人胰腺癌细胞(PANC-1和Mia Pa Ca-2)中TCP1α蛋白基因,Western blot检测3种TCP1α蛋白(TCP1α-1、TCP1α-2、TCP1α-3)表达情况,验证SiRNA沉默效果。以体外增殖实验(MTT实验)和Transwell迁移侵袭实验验证抑制TCP1α蛋白表达对两种胰腺癌细胞体外增殖、迁移及侵袭等生物学行为的影响。Western blot检测PD98059和CHX共同处理的基因沉默人胰腺癌细胞的TCPIα蛋白表达情况。结果:沉默TCP1α基因后两种人胰腺癌细胞增殖和迁移侵袭能力显著降低。结论:抑制TCP1α蛋白表达能明显降低人胰腺导管腺癌细胞的增殖侵袭及迁移能力,且能诱发人胰腺导管腺癌细胞的急性凋亡。
基金Supported by Beijing Key Laboratory Special Fund,No.Z141107004414042
文摘AIM:To investigate the value of chaperonin containing TCP1,subunit 3(CCT3) to predict the prognosis of patients with hepatocellular carcinoma(HCC) and determine its function in HCC progression.METHODS:CCT3 expression levels were examined in human non-cancerous liver tissues and a variety of HCC cell lines by quantitative real-time PCR and immunoblotting.CCT3 expression was suppressed by small interfering RNA.The effects of reducing CCT3 expression in HCC cells were tested.The3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide(MTT) assay,cell counting experiment,cell cycle assay,apoptosis assay and invasion assay were employed to evaluate cell functions in vitro.Immunohistochemistry was performed on HCC specimens.In addition,CCT3 expression in HCC specimens was also assessed at the protein and mRNA level.Associations between clinicopathological characteristics and prognosis were analyzed,along with the possible mechanisms involved in CCT3's function in HCC progression.RESULTS:The expression levels of CCT3 mRNA and protein were upregulated in HCC cell lines in contrast to adjacent non-cancerous tissues.Reducing CCT3 expression not only suppressed cell proliferation in cell counts,MTT assay,cell cycle assay and induced cell apoptosis(P < 0.05 vs negative control),but also inhibited the tumor cell invasion capacity in vitro {P< 0.01 vs negative control).Overexpression of CCT3 in the nuclei of cancer cells in HCC specimens(58of 104 patients,55.8%) was associated with poor prognosis in HCC patients(3-year survival rate,55.5%vs 84.2%,P = 0.020) after hepatectomy.Mechanistic analyses showed that signal transducer and activator of transcription 3(STAT3) activation was decreased even when stimulated by interleukin-6 after knocking down CCT3 in the HepG2 cell line.CONCLUSION:Overexpression of CCT3 in the nuclei of cancerous cells is associated with HCC progression.CCT3 may be a target that affects the activation of STAT3 in HCC.
基金supported by the National Key R&D Program of China(2020YFE0202900)the National Natural Science Foundation of China(31972460,32160729)+2 种基金the China Agriculture Research System of MOF and MARA(CARS-19)Science and Technology Support Project of Guizhou Province(ZK[2021]154)the Priority Academic Program Development of Jiangsu Higher Education Institutions.
文摘TCP transcription factors are essential for plant growth and environmental response,but little is known about their specific roles in growth,hormone response,and stress response in tea plants.Here,CsTCP5 and CsTCP18 were cloned from the tea variety'Longjing 43',and subcellular localization revealed that both CsTCP5 and CsTCP18 proteins were present in the nucleus.Exogenous abscisic acid(ABA)and methyl jasmonate(MeJA)had different effects on CsTCP5 and CsTCP18 expression,but both genes were repressed by drought and low-temperature treatment.Yeast-two hybrid(Y2H)and bimolecular fluorescence complementation(BiFC)assays demonstrated that neither transcription factor was transcriptionally active but that each interacted with members of the CsMYB and CsJAZ families.Transgenic Arabidopsis plants overexpressing CsTCP5 had smaller cotyledons and fewer lateral roots compared with wild-type(WT)and empty vector(EV)Arabidopsis,and their root growth and germination rate were inhibited by ABA and MeJA treatment.Lateral root numbers also decreased significantly in CsTCP18-OE Arabidopsis after MeJA treatment.These results demonstrate that CsTCP5 and CsTCP18 have regulatory effects on cotyledon development,lateral root growth,and germination rate,and these effects can be modulated by ABA and MeJA.Under drought stress,the CsTCP5-OE and CsTCP18-OE lines exhibited greater survival,lower plant height,smaller rosette leaves,delayed flowering,increased activities of antioxidant enzymes,decreased MDA content,and increased proline content compared with WT and EV Arabidopsis.These findings suggest that CsTCP5 and CsTCP18 are important for tea plant growth,interact with ABA and MeJA pathways,and play roles in stress response.
文摘(Molecular Plant8(3):482–485;March 2015;https://doi.org/10.1016/j.molp.2014.11.018)The supplemental figure S3(panel A)was amended since the picture of the yeast control spot TCP14-AD/M5RGL2BD(-W-L medium)has been used in place of TCP15-AD/M5GAI-BD.The authors apologize for not detecting this error prior to publication and for any inconvenience that may have caused.
基金supported by the Joint Funds for the Innovation of Science and Technology,Fujian province(2017Y9054 and 2017Y9100)the Special Financial Found of Fujian Provincial Department of Science and Technology(Min 2015-1297)+2 种基金the Construction Project of Fujian Medical Center of Hematology(Min 2017-04)the National and Fujian Provincial Key Clinical Specialty Discipline Construction Program of Chinaand the Foundation of Fujian Key Laboratory of Hematology(2009J1004).
文摘Dear Editor,Molecular chaperones are critical mediators of oncogenesis and necessary for cell survival.1 The cytoplasmic chaperone TRiC(Tcomplex protein-1 ring complex,also known as CCT)comprises two back-to-back stacked rings,with each ring containing eight subunits(CCT1–CCT8).2 And the effects of CCT subunits on tumors may be different.However,the roles of the CCT1 subunit(also known as TCP1)in hepatocellular carcinoma(HCC)and its molecular mechanism have not been investigated thoroughly.Understanding these details can provide new ideas and strategies for treating HCC.
基金Supported by the National Natural Science Foundation of China,No.82160213 and No.U22A2022the Guangxi Natural Science Foundation,No.2023GXNSFAA026029,No.2024GXNSFBA010059,and No.2024GXNSFAA010079。
文摘BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 subunit 3(CCT3)expression and its clinical implications,and its effects on LSCC cell growth.METHODS Systematic data on CCT3 mRNA expression were collected from biomedical databases,and integrated further based on the standardized mean difference and the summary receiver operating characteristic curve.Single-cell RNA-seq data were mined to validate the expression level of CCT3 mRNA.In-house immunohistochemistry was performed to explore the CCT3 protein levels of clinical LSCC samples and their relationship with clinical parameters.The growth function of LSCC cell was analyzed using CRISPR knockout screening.CCT3-related signaling pathway analyses were conducted using gene set enrichment analysis.Protein-protein interaction network construction was performed to identify hub genes.RESULTS CCT3 mRNA was significantly overexpressed in 269 LSCC tissues cases across multiple independent datasets(standardized mean difference=32,area under the curve=0.93);At the translational level,the in-house immunohistochemical analysis further demonstrated the consistent upregulation of CCT3 protein in 88 cases of LSCC samples(58 non-LSCC samples vs 30 LSCC samples,P=1.4e^(-14)).Analysis of clinical parameters showed no significant differences among subgroup.Functional characterization with clustered regularly interspaced short palindromic repeats--mediated gene knockout revealed that depletion of CCT3 potently suppressed LSCC cell viability in vitro.Gene set enrichment analysis indicated that CCT3 was markedly associated with several key oncogenic pathways,including extracellular matrix receptor interaction and cell cycle regulation pathways.CONCLUSION CCT3 upregulation in LSCC may influence cellular growth by regulating related pathways,indicating its potential as a biomarker and therapeutic target for LSCC.
