目的检测动力蛋白激活蛋白2(dynactin 2,DCTN2)和肿瘤高表达细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)在胃癌组织和癌旁组织中的表达差异性,探讨DCTN2和CREPT表达与胃癌患者的临床病理特...目的检测动力蛋白激活蛋白2(dynactin 2,DCTN2)和肿瘤高表达细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)在胃癌组织和癌旁组织中的表达差异性,探讨DCTN2和CREPT表达与胃癌患者的临床病理特征及预后的关系。方法纳入2014年3月~2015年9月于徐州医科大学附属医院行胃癌切除术并经组织病理学证实为胃癌的患者90例,采用免疫组织化学法检测DCTN2和CREPT在胃癌组织及癌旁组织中的表达水平,采用Spearman法分析胃癌组织中DCTN2与CREPT的表达相关性;分析DCTN2与CREPT表达水平与胃癌患者临床病理特征的关系;采用Kaplan-Meier法及Log-rank法分析DCTN2和CREPT的表达水平对胃癌患者生存期的影响;通过单因素及多因素COX回归分析影响胃癌患者预后的独立危险因素。结果胃癌组织中DCTN2和CREPT的表达水平显著高于癌旁组织,且两者的表达呈正相关(P<0.05);DCTN2和CREPT的表达水平与胃癌分化程度、TNM分期、肿瘤浸润深度、淋巴结转移情况显著相关(P<0.05),与患者的性别、年龄、远处转移情况无明显相关性(P>0.05);预后分析结果显示,DCTN2高表达组胃癌患者的术后1、3、5年累积生存率及总生存率均显著低于DCTN2低表达组,CREPT高表达组胃癌患者的术后5年累积生存率和总生存率均显著低于CREPT低表达组(P<0.05)。单因素及多因素COX回归分析结果显示,DCTN2和CREPT均是影响胃癌患者预后的独立危险因素。结论DCTN2和CREPT在胃癌组织中均显著高表达,其高表达与胃癌患者的不良预后密切相关,有望成为胃癌诊断的潜在标志物和治疗新靶点,并有助于判断胃癌患者的预后。展开更多
目的探讨CREPT(cell-cycle related and expression-elevated protein in tumor)与Cyclin D1在肾透明细胞癌(RCCC)及癌旁组织的表达情况。方法采用实时荧光定量PCR检测40例RCCC及其相应癌旁组织中CREPT和Cyclin D1的mRNA表达,采用Wester...目的探讨CREPT(cell-cycle related and expression-elevated protein in tumor)与Cyclin D1在肾透明细胞癌(RCCC)及癌旁组织的表达情况。方法采用实时荧光定量PCR检测40例RCCC及其相应癌旁组织中CREPT和Cyclin D1的mRNA表达,采用Western blot检测20例RCCC及其相应癌旁组织中CREPT和Cyclin D1蛋白表达。结果荧光定量PCR相对定量发现,CREPT和Cyclin D1在RCCC组织中的表达明显高于相应癌旁组织(1.000±0.000 vs 3.034±1.868,P=0.000;1.000±0.000 vs 5.611±4.089,P=0.000);Western blot检测显示CREPT蛋白和Cyclin D1蛋白在RCCC组织中的表达显著高于癌旁组织(0.618±0.139 vs1.062±0.090,P=0.000;0.712±0.165 vs 1.066±0.165,P=0.000);在RCCC中CREPT与Cyclin D1的mRNA及蛋白表达水平均呈正相关性(r=0.875,P=0.000;r=0.786,P=0.000)。结论 CREPT在RCCC中过表达提示该基因可能为RCCC的癌基因,可能作为未来RCCC诊断的标志物和基因治疗的靶点。展开更多
目的:检测CREPT(cell-cycle related and expression-elevated protein in tumor)与细胞周期蛋白D(CyclinD3)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,分析二者在促进NSCLC发生发展过程中的作用。方法:免疫组织化学...目的:检测CREPT(cell-cycle related and expression-elevated protein in tumor)与细胞周期蛋白D(CyclinD3)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,分析二者在促进NSCLC发生发展过程中的作用。方法:免疫组织化学法检测CREPT、CyclinD3在271例NSCLC及相应正常组织中的表达情况,统计学分析二者间的相关性及其在临床病理因素中的差异性。结果:CREPT和CyclinD3定位于细胞核与细胞质中,在NSCLC组织中高表达,癌组织与正常组织中阳性表达率差异性极显著(P<0.001)。CREPT、CyclinD3在腺癌、鳞癌间表达无统计学差异(P=0.638,P=0.503);在总NSCLC、腺癌、鳞癌的病理分级间,表达均具有极显著性差异(P<0.001);在总NSCLC组织TNM分期、淋巴结是否转移间的表达差异性显著(P=0.025,P=0.001;P=0.003,P=0.026);在总NSCLC、腺癌、鳞癌中,CREPT和CyclinD3表达秩相关系数r分别是0.458、0.393、0.468;P<0.001,二者具有正相关性。结论:CREPT和CyclinD3是NSCLC组织与相应正常组织表达的差异蛋白,二者的过表达可能在NSCLC的发生发展过程中起作用。展开更多
CREPT是一种新近发现的癌基因。研究显示,CREPT在肿瘤的发生发展起到了重要的作用,它能够调控许多与肿瘤发生发展相关基因的表达,能够增强细胞的增殖和迁移能力。抑制CREPT的表达后,肿瘤细胞的生长、迁移能力受到抑制,并能够诱导细胞的...CREPT是一种新近发现的癌基因。研究显示,CREPT在肿瘤的发生发展起到了重要的作用,它能够调控许多与肿瘤发生发展相关基因的表达,能够增强细胞的增殖和迁移能力。抑制CREPT的表达后,肿瘤细胞的生长、迁移能力受到抑制,并能够诱导细胞的凋亡。研究还发现,CREPT表达与肿瘤的临床指标密切相关,如病理分级、淋巴结转移、复发及生存期等。CREPT包括RPR结构域(regulation of nuclear premRNA domain,RPR),能够与RNA聚合酶Ⅱ相互作用进一步影响下游基因转录调控,提示它可能是很有潜力的药物治疗靶点。本文就CREPT的近年来的研究进展及其与肿瘤的关系进行综述。展开更多
目的探讨肾癌组织中肿瘤细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)的表达与临床病理特征和生存率的关系。方法纳入90例于2014至2016年间在北京大学人民医院接受了根治性肾切除术并经组织...目的探讨肾癌组织中肿瘤细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)的表达与临床病理特征和生存率的关系。方法纳入90例于2014至2016年间在北京大学人民医院接受了根治性肾切除术并经组织学证实的肾癌患者,运用免疫组织化学方法检测这些患者的癌组织和癌旁组织中CREPT的表达,并结合临床病理资料分析CREPT表达水平与患者TNM分期和Fuhrman分级的关系。通过Kaplan-Meier生存分析及多因素COX回归分析CREPT表达与预后的关系。结果46.7%(42/90)的患者癌组织中CREPT表达为高水平,而在癌旁组织中所有患者CREPT表达均为低水平,组间比较差异有统计学意义(P<0.05)。CREPT的表达水平与TNM分期(P=0.001)和Fuhrman分级(P<0.001)有关联,但与性别(P=0.149)、年龄(P=0.605)、肿瘤大小(P=0.673)和组织学类型(P=0.756)无关。截至2018年12月,有85例患者完成随访。Kaplan-Meier分析显示,CREPT高表达患者总生存时间和无瘤生存时间均低于CREPT低表达患者,差异有统计学意义(P<0.05)。多因素COX回归分析显示,患者总生存时间及无瘤生存时间与CREPT的表达水平有关(P<0.05)。结论CREPT在肾癌组织中呈高表达,表达水平与临床分期、分级以及患者预后密切相关。展开更多
AIM To investigate expression of cell cycle-related and expression-elevated protein in tumor(CREPT) in colorectal cancer(CRC) and determine its prognostic value in response to 5-fluorouracil(5-FU).METHODS The relative...AIM To investigate expression of cell cycle-related and expression-elevated protein in tumor(CREPT) in colorectal cancer(CRC) and determine its prognostic value in response to 5-fluorouracil(5-FU).METHODS The relative expression of CREPT in CRC tumor samples was determined using immunohistochemistry. The protein content in cell lines was analyzed by immunoblotting. Cell viability was measured with the CCK-8 assay. Cell cycle and apoptosis analyses were performed with flow cytometry.RESULTS CREPT was overexpressed in CRC tissues and correlated with histological grade. Clinicopathological analysis indicated that CREPT was positively related to tumor progression. Exogenous expression of CREPT stimulated cell proliferation and accelerated the cell cycle. More importantly, high expression of CREPT sensitized CRC cells to 5-FU treatment. Furthermore, we demonstrated that 5-FU elicited significant apoptosis in CREPT-positive cells.CONCLUSION Aberrant overexpression of CREPT contributes to tumorigenesis of CRC by promoting cell proliferation and accelerating the cell cycle, and confers sensitivity to 5-FU. CREPT is a potential prognostic biomarker for 5-FU in CRC.展开更多
CREPT and p15RS are two recently identified homologous proteins that regulate cell proliferation in an opposite way and are closely related to human cancer development.Both CREPT and p15RS consist of an N-terminal RPR...CREPT and p15RS are two recently identified homologous proteins that regulate cell proliferation in an opposite way and are closely related to human cancer development.Both CREPT and p15RS consist of an N-terminal RPR domain and a C-terminal domain with high sequence homology.The transcription enhancement by CREPT is attributed to its interaction with RNA polymerase II(Pol II).Here we provide biochemical and structural evidence to support and extend this molecular mechanism.Through fluorescence polarization analysis,we show that the RPR domains of CREPT and p15RS(CREPT-RPR and p15RS-RPR)bind to different Pol II C-terminal domain(CTD)phosphoisoforms with similar affinity and specificity.We also determined the crystal structure of p15RS-RPR.Sequence and structural comparisons with RPR domain of Rtt103,a homolog of CREPT and p15RS in yeast,reveal structural basis for the similar binding profile of CREPT-RPR and p15RS-RPR with Pol II CTD.We also determined the crystal structure of the C-terminal domain of CREPT(CREPT-CTD),which is a long rod-like dimer and each monomer adopts a coiled-coil structure.We propose that dimerization through the C-terminal domain enhances the binding strength between CREPT or p15RS with Pol II by increasing binding avidity.Our results collectively reveal the respective roles of N-terminal RPR domain and C-terminal domain of CREPT and p15RS in recognizing RNA Pol II.展开更多
文摘目的检测动力蛋白激活蛋白2(dynactin 2,DCTN2)和肿瘤高表达细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)在胃癌组织和癌旁组织中的表达差异性,探讨DCTN2和CREPT表达与胃癌患者的临床病理特征及预后的关系。方法纳入2014年3月~2015年9月于徐州医科大学附属医院行胃癌切除术并经组织病理学证实为胃癌的患者90例,采用免疫组织化学法检测DCTN2和CREPT在胃癌组织及癌旁组织中的表达水平,采用Spearman法分析胃癌组织中DCTN2与CREPT的表达相关性;分析DCTN2与CREPT表达水平与胃癌患者临床病理特征的关系;采用Kaplan-Meier法及Log-rank法分析DCTN2和CREPT的表达水平对胃癌患者生存期的影响;通过单因素及多因素COX回归分析影响胃癌患者预后的独立危险因素。结果胃癌组织中DCTN2和CREPT的表达水平显著高于癌旁组织,且两者的表达呈正相关(P<0.05);DCTN2和CREPT的表达水平与胃癌分化程度、TNM分期、肿瘤浸润深度、淋巴结转移情况显著相关(P<0.05),与患者的性别、年龄、远处转移情况无明显相关性(P>0.05);预后分析结果显示,DCTN2高表达组胃癌患者的术后1、3、5年累积生存率及总生存率均显著低于DCTN2低表达组,CREPT高表达组胃癌患者的术后5年累积生存率和总生存率均显著低于CREPT低表达组(P<0.05)。单因素及多因素COX回归分析结果显示,DCTN2和CREPT均是影响胃癌患者预后的独立危险因素。结论DCTN2和CREPT在胃癌组织中均显著高表达,其高表达与胃癌患者的不良预后密切相关,有望成为胃癌诊断的潜在标志物和治疗新靶点,并有助于判断胃癌患者的预后。
文摘目的探讨CREPT(cell-cycle related and expression-elevated protein in tumor)与Cyclin D1在肾透明细胞癌(RCCC)及癌旁组织的表达情况。方法采用实时荧光定量PCR检测40例RCCC及其相应癌旁组织中CREPT和Cyclin D1的mRNA表达,采用Western blot检测20例RCCC及其相应癌旁组织中CREPT和Cyclin D1蛋白表达。结果荧光定量PCR相对定量发现,CREPT和Cyclin D1在RCCC组织中的表达明显高于相应癌旁组织(1.000±0.000 vs 3.034±1.868,P=0.000;1.000±0.000 vs 5.611±4.089,P=0.000);Western blot检测显示CREPT蛋白和Cyclin D1蛋白在RCCC组织中的表达显著高于癌旁组织(0.618±0.139 vs1.062±0.090,P=0.000;0.712±0.165 vs 1.066±0.165,P=0.000);在RCCC中CREPT与Cyclin D1的mRNA及蛋白表达水平均呈正相关性(r=0.875,P=0.000;r=0.786,P=0.000)。结论 CREPT在RCCC中过表达提示该基因可能为RCCC的癌基因,可能作为未来RCCC诊断的标志物和基因治疗的靶点。
文摘目的:检测CREPT(cell-cycle related and expression-elevated protein in tumor)与细胞周期蛋白D(CyclinD3)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,分析二者在促进NSCLC发生发展过程中的作用。方法:免疫组织化学法检测CREPT、CyclinD3在271例NSCLC及相应正常组织中的表达情况,统计学分析二者间的相关性及其在临床病理因素中的差异性。结果:CREPT和CyclinD3定位于细胞核与细胞质中,在NSCLC组织中高表达,癌组织与正常组织中阳性表达率差异性极显著(P<0.001)。CREPT、CyclinD3在腺癌、鳞癌间表达无统计学差异(P=0.638,P=0.503);在总NSCLC、腺癌、鳞癌的病理分级间,表达均具有极显著性差异(P<0.001);在总NSCLC组织TNM分期、淋巴结是否转移间的表达差异性显著(P=0.025,P=0.001;P=0.003,P=0.026);在总NSCLC、腺癌、鳞癌中,CREPT和CyclinD3表达秩相关系数r分别是0.458、0.393、0.468;P<0.001,二者具有正相关性。结论:CREPT和CyclinD3是NSCLC组织与相应正常组织表达的差异蛋白,二者的过表达可能在NSCLC的发生发展过程中起作用。
文摘CREPT是一种新近发现的癌基因。研究显示,CREPT在肿瘤的发生发展起到了重要的作用,它能够调控许多与肿瘤发生发展相关基因的表达,能够增强细胞的增殖和迁移能力。抑制CREPT的表达后,肿瘤细胞的生长、迁移能力受到抑制,并能够诱导细胞的凋亡。研究还发现,CREPT表达与肿瘤的临床指标密切相关,如病理分级、淋巴结转移、复发及生存期等。CREPT包括RPR结构域(regulation of nuclear premRNA domain,RPR),能够与RNA聚合酶Ⅱ相互作用进一步影响下游基因转录调控,提示它可能是很有潜力的药物治疗靶点。本文就CREPT的近年来的研究进展及其与肿瘤的关系进行综述。
文摘目的探讨肾癌组织中肿瘤细胞周期相关蛋白(cell-cycle-related and expression-elevated protein in tumor,CREPT)的表达与临床病理特征和生存率的关系。方法纳入90例于2014至2016年间在北京大学人民医院接受了根治性肾切除术并经组织学证实的肾癌患者,运用免疫组织化学方法检测这些患者的癌组织和癌旁组织中CREPT的表达,并结合临床病理资料分析CREPT表达水平与患者TNM分期和Fuhrman分级的关系。通过Kaplan-Meier生存分析及多因素COX回归分析CREPT表达与预后的关系。结果46.7%(42/90)的患者癌组织中CREPT表达为高水平,而在癌旁组织中所有患者CREPT表达均为低水平,组间比较差异有统计学意义(P<0.05)。CREPT的表达水平与TNM分期(P=0.001)和Fuhrman分级(P<0.001)有关联,但与性别(P=0.149)、年龄(P=0.605)、肿瘤大小(P=0.673)和组织学类型(P=0.756)无关。截至2018年12月,有85例患者完成随访。Kaplan-Meier分析显示,CREPT高表达患者总生存时间和无瘤生存时间均低于CREPT低表达患者,差异有统计学意义(P<0.05)。多因素COX回归分析显示,患者总生存时间及无瘤生存时间与CREPT的表达水平有关(P<0.05)。结论CREPT在肾癌组织中呈高表达,表达水平与临床分期、分级以及患者预后密切相关。
基金Supported by The Key Project Grant from the National Natural Science Foundation of China,No.81372167,No.81572729,No.81402293,No.81372372 and No.81230044The National Key Research and Development Program of China,No.2016YFA0500301
文摘AIM To investigate expression of cell cycle-related and expression-elevated protein in tumor(CREPT) in colorectal cancer(CRC) and determine its prognostic value in response to 5-fluorouracil(5-FU).METHODS The relative expression of CREPT in CRC tumor samples was determined using immunohistochemistry. The protein content in cell lines was analyzed by immunoblotting. Cell viability was measured with the CCK-8 assay. Cell cycle and apoptosis analyses were performed with flow cytometry.RESULTS CREPT was overexpressed in CRC tissues and correlated with histological grade. Clinicopathological analysis indicated that CREPT was positively related to tumor progression. Exogenous expression of CREPT stimulated cell proliferation and accelerated the cell cycle. More importantly, high expression of CREPT sensitized CRC cells to 5-FU treatment. Furthermore, we demonstrated that 5-FU elicited significant apoptosis in CREPT-positive cells.CONCLUSION Aberrant overexpression of CREPT contributes to tumorigenesis of CRC by promoting cell proliferation and accelerating the cell cycle, and confers sensitivity to 5-FU. CREPT is a potential prognostic biomarker for 5-FU in CRC.
基金supported by Ministry of Science and Technology(2010CB912402 and 2011CB910502)Ministry of Health(2012ZX1000-1009) and the Fok Ying Tung Education Foundation to Wang XinQuan+1 种基金the National Natural Science Foundation of China(81230044,81372167 and 31071225)the Tsinghua Science Foundation(20121080018)to Chang ZhiJie
文摘CREPT and p15RS are two recently identified homologous proteins that regulate cell proliferation in an opposite way and are closely related to human cancer development.Both CREPT and p15RS consist of an N-terminal RPR domain and a C-terminal domain with high sequence homology.The transcription enhancement by CREPT is attributed to its interaction with RNA polymerase II(Pol II).Here we provide biochemical and structural evidence to support and extend this molecular mechanism.Through fluorescence polarization analysis,we show that the RPR domains of CREPT and p15RS(CREPT-RPR and p15RS-RPR)bind to different Pol II C-terminal domain(CTD)phosphoisoforms with similar affinity and specificity.We also determined the crystal structure of p15RS-RPR.Sequence and structural comparisons with RPR domain of Rtt103,a homolog of CREPT and p15RS in yeast,reveal structural basis for the similar binding profile of CREPT-RPR and p15RS-RPR with Pol II CTD.We also determined the crystal structure of the C-terminal domain of CREPT(CREPT-CTD),which is a long rod-like dimer and each monomer adopts a coiled-coil structure.We propose that dimerization through the C-terminal domain enhances the binding strength between CREPT or p15RS with Pol II by increasing binding avidity.Our results collectively reveal the respective roles of N-terminal RPR domain and C-terminal domain of CREPT and p15RS in recognizing RNA Pol II.
