目的探讨出核因子(chromosome maintenance region 1,CRM1)抑制剂KPT-185对人前列腺癌细胞生长、凋亡、迁徙的影响及影响产生的原因。方法采用MTT、细胞集落形成能力测定法检测KPT-185对前列腺癌细胞生长的影响;划痕愈合实验检测KPT-18...目的探讨出核因子(chromosome maintenance region 1,CRM1)抑制剂KPT-185对人前列腺癌细胞生长、凋亡、迁徙的影响及影响产生的原因。方法采用MTT、细胞集落形成能力测定法检测KPT-185对前列腺癌细胞生长的影响;划痕愈合实验检测KPT-185对前列腺癌细胞迁徙能力的影响;流式细胞仪检测KPT-185对细胞周期的影响;人组蛋白(Histone)/DNA酶联免疫法和膜联蛋白V-FITC/PI双染法流式细胞术检测细胞凋亡;免疫印迹检测蛋白表达。结果 KPT-185能极大地抑制人前列腺癌细胞的生长,使其细胞周期阻滞于G0~G1期,并导致细胞凋亡。KPT-185引起的细胞生长抑制和凋亡是通过其抑制CRM1的核输出,使抑癌蛋白p53和p21保留于核内而发挥作用的。结论KPT-185是一种新型的CRM1抑制剂,有望成为治疗前列腺癌的候选药物。展开更多
Mantle cell lymphoma （MCL） is an aggressive histotype of B-cell non-Hodgkin lymphoma. The disease has no known cure, which prompts the urgent need for novel therapeutic agents. Chromosomal region maintenance 1 （CRM...Mantle cell lymphoma （MCL） is an aggressive histotype of B-cell non-Hodgkin lymphoma. The disease has no known cure, which prompts the urgent need for novel therapeutic agents. Chromosomal region maintenance 1 （CRM1） may play a role in human neoplasia and serve as a novel target of cancer treatment. This study summarizes MCL pathogenesis and determines the involvement of CRM1 in the regulation of several vital signaling pathways contributing to MCL pathogenesis, including the pathways of cell cycle progression, DNA damage response, phosphoinositide kinase-3, nuclear factor-kB activation, and chromosomal stability. A preclinical study is also presented to compare the CRNI1 status in MCL cell lines and primary MCL cells with normal B cells, as well as the therapeutic efficiency of CRM1 inhibition in MCL in vitro and in vivo, which make these agents potential targets of novel MCL treatments.展开更多
目的:研究靶向调控运输蛋白(Chromosome region maintenance1,CRM1)在创伤性脑损伤中的表达变化。方法:采用Western blot和免疫组织化学荧光法检测脑损伤后CRM1在脑中的表达变化。结果:(1)假手术组皮质CRM1蛋白水平低,在损伤后12小时逐...目的:研究靶向调控运输蛋白(Chromosome region maintenance1,CRM1)在创伤性脑损伤中的表达变化。方法:采用Western blot和免疫组织化学荧光法检测脑损伤后CRM1在脑中的表达变化。结果:(1)假手术组皮质CRM1蛋白水平低,在损伤后12小时逐步增加,在损伤后3天达到高峰,之后恢复到正常水平。非手术对侧皮层中CRM1表达没有明显改变。(2)免疫组化荧光测定:损伤后CRM1表达水平不但上升,且有入核的亚定位变化,主要定位在神经元中。而假手术组和对侧则主要定位在胞浆中。结论:推测CRM1在创伤性脑损伤后与神经元凋亡有关。展开更多
AIM To evaluate the predictive value of the expression of chromosomal maintenance(CRM)1 and cyclindependent kinase(CDK)5 in gastric cancer(GC) patients after gastrectomy.METHODS A total of 240 GC patients who received...AIM To evaluate the predictive value of the expression of chromosomal maintenance(CRM)1 and cyclindependent kinase(CDK)5 in gastric cancer(GC) patients after gastrectomy.METHODS A total of 240 GC patients who received standard gastrectomy were enrolled in the study. The expression level of CRM1 and CDK5 was detected by immunohistochemistry. The correlations between CRM1 and CDK5 expression and clinicopathological factors were explored. Univariate and multivariate survival analyses were used to identify prognostic factors for GC. Receiver operating characteristic analysis was used to compare the accuracy of the prediction of clinical outcome by the parameters. RESULTS The expression of CRM1 was significantly related to size of primary tumor(P = 0.005), Borrmann type(P = 0.006), degree of differentiation(P = 0.004), depth of invasion(P = 0.008), lymph node metastasis(P = 0.013), TNM stage(P = 0.002) and distant metastasis(P = 0.015). The expression of CDK5 was significantly related to sex(P = 0.048) and Lauren's classification(P = 0.011). Multivariate Cox regression analysis identified that CRM1 and CDK5 co-expression status was an independent prognostic factor for overall survival(OS) of patients with GC. Integration of CRM1 and CDK5 expression could provide additional prognostic value for OS compared with CRM1 or CDK5 expression alone(P = 0.001).CONCLUSION CRM1 and CDK5 co-expression was an independent prognostic factors for GC. Combined CRM1 and CDK5 expression could provide a prognostic model for OS of GC.展开更多
目的:研究NF-κBp50、JAB1和CRM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及其意义。方法:采用免疫组化方法检测94例DLBCL中NF-κBp50、JAB1和CRM1的表达。结果:NF-κBp50、JAB1和CRM1在94例DLBCL的阳性表...目的:研究NF-κBp50、JAB1和CRM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及其意义。方法:采用免疫组化方法检测94例DLBCL中NF-κBp50、JAB1和CRM1的表达。结果:NF-κBp50、JAB1和CRM1在94例DLBCL的阳性表达率分别为70.2%(66/94)、77.7%(73/94)和69.1%(65/94)。NF-κBp50在DL-BCL中非生发中心型(non-GCB)中表达明显高于生发中心型(GCB),差异有统计学意义(P<0.01),但JAB1、CRM1与DLBCL分型无关(P>0.05)。NF-κBp50、JAB1和CRM1与DLBCL患者性别、年龄、发病部位均无关(P>0.05)。DL-BCL中NF-κBp50与JAB1、NF-κBp50与CRM1的表达均呈正相关(rs=0.209,P<0.05;rs=0.270,P<0.01)。结论:NF-κp50在DLBCL不同亚型之间表达的差异,可能有助于DLBCL亚型的诊断和预后的判断。JAB1和CRM1对NF-κBp50表达具有正向调节作用。展开更多
Cellular homeostasis crucially relies on the correct nucleocytoplasmic distribution of a vast number of proteins and RNA molecules,which are shuttled in and out of the nucleus by specialized transport receptors.The nu...Cellular homeostasis crucially relies on the correct nucleocytoplasmic distribution of a vast number of proteins and RNA molecules,which are shuttled in and out of the nucleus by specialized transport receptors.The nuclear export receptor XPO1,also called CRM1,mediates the translocation of hundreds of proteins and several classes of RNA to the cytoplasm,and thus regulates critical signaling pathways and cellular functions.The normal function of XPO1 appears to be often disrupted in malignant cells due to gene mutations or,most commonly,aberrant overexpression.Due to its important physiological roles and its frequent alteration in human tumors,XPO1 is a promising target for cancer therapy.XPO1 inhibitors have undergone extensive testing as therapeutic agents in preclinical models of cancer,with promising results.One of these inhibitors,Selinexor,is currently being evaluated in multiple clinical trials of different types of solid tumors and hematological malignancies.Here,we review several key aspects of XPO1 function,as well as the mechanisms that may lead to its alteration in cancer,and provide an update on the status of XPO1 inhibitors being developed as drugs for cancer therapy,including the definitive results of the first clinical trials with Selinexor that have been recently published.展开更多
Objectives: Multi-drug resistance (MDR) to chemotherapy remains a major obstacle to overcome in the successful treatment of patients with cancers. It was recently discovered that Leptomycin B (LMB) reduces the paclita...Objectives: Multi-drug resistance (MDR) to chemotherapy remains a major obstacle to overcome in the successful treatment of patients with cancers. It was recently discovered that Leptomycin B (LMB) reduces the paclitaxel-induced MDR in CCRF-CEM/Taxol cells. However, the mechanism remains unclear. Here we sought to explore the mechanism of LMB to reduce the MDR induced by paclitaxel. Results: LMB has remarkable cytotoxic effects in both sensitive CCRF-CEM and resistant CCRF-CEM/Taxol cell lines. The paclitaxel-induced MDR was reduced by 0.013 μm of LMB. Lower concentration of LMB regulated cell cycle progress, in situ expressions of P-gp, MRP1, and LRP, expression of CRM1, and localization of P-gp and CRM1 in CCRF-CEM/taxol cells. Study Design: Cytotoxicity of LMB on cancerous cell lines was determined by MTT assay. Cell cycle progress and in situ expressions of P-gp, MRP1, and LRP were analyzed by flow cytometry. Expression of CRM1 in the cells was examined by Western blot. And co-localization between P-gp and CRM1 was determined by laser confocal microscopy. Conclusion: The paclitaxel-induced MDR of CCRFCEM/Taxol cells was reduced by lower concentration of LMB. The mechanisms might be related to decreasing in situ expression of drug transporter proteins, promoting cell cycle progress, and altering co-localization between P-gp and CRM1 in the resistant cells.展开更多
The function of the herpes simplex virus type 1 （HSV-1） UL4 protein is still elusive. Our objective is to investigate the subcellular transport mechanism of the UL4 protein. In this study, fluorescence microscopy wa...The function of the herpes simplex virus type 1 （HSV-1） UL4 protein is still elusive. Our objective is to investigate the subcellular transport mechanism of the UL4 protein. In this study, fluorescence microscopy was employed to investigate the subcellular localization of UL4 and characterize the transport mechanism in living cells. By constructing a series of deletion mutants fused with enhanced yellow fluorescent protein （EYFP）, the nuclear export signals （NES） of UL4 were for the first time mapped to amino acid residues 178 to 186. In addition, the N-terminal 19 amino acids are identified to be required for the granule-like cytoplasmic pattem of UL4. Furthermore, the UL4 protein was demonstrated to be exported to the cytoplasm through the NES in a chromosomal region maintenance 1 （CRM1）-dependent manner involving RanGTP hydrolysis展开更多
文摘目的探讨出核因子(chromosome maintenance region 1,CRM1)抑制剂KPT-185对人前列腺癌细胞生长、凋亡、迁徙的影响及影响产生的原因。方法采用MTT、细胞集落形成能力测定法检测KPT-185对前列腺癌细胞生长的影响;划痕愈合实验检测KPT-185对前列腺癌细胞迁徙能力的影响;流式细胞仪检测KPT-185对细胞周期的影响;人组蛋白(Histone)/DNA酶联免疫法和膜联蛋白V-FITC/PI双染法流式细胞术检测细胞凋亡;免疫印迹检测蛋白表达。结果 KPT-185能极大地抑制人前列腺癌细胞的生长,使其细胞周期阻滞于G0~G1期,并导致细胞凋亡。KPT-185引起的细胞生长抑制和凋亡是通过其抑制CRM1的核输出,使抑癌蛋白p53和p21保留于核内而发挥作用的。结论KPT-185是一种新型的CRM1抑制剂,有望成为治疗前列腺癌的候选药物。
基金supported in part by grants from Fujian Provincial Department of Science & Technology(2009-CXB-57/ 2011J01252)Bureau of Science & Technology of Xiamen,China (3502Z20094012)
文摘Mantle cell lymphoma （MCL） is an aggressive histotype of B-cell non-Hodgkin lymphoma. The disease has no known cure, which prompts the urgent need for novel therapeutic agents. Chromosomal region maintenance 1 （CRM1） may play a role in human neoplasia and serve as a novel target of cancer treatment. This study summarizes MCL pathogenesis and determines the involvement of CRM1 in the regulation of several vital signaling pathways contributing to MCL pathogenesis, including the pathways of cell cycle progression, DNA damage response, phosphoinositide kinase-3, nuclear factor-kB activation, and chromosomal stability. A preclinical study is also presented to compare the CRNI1 status in MCL cell lines and primary MCL cells with normal B cells, as well as the therapeutic efficiency of CRM1 inhibition in MCL in vitro and in vivo, which make these agents potential targets of novel MCL treatments.
文摘目的:研究靶向调控运输蛋白(Chromosome region maintenance1,CRM1)在创伤性脑损伤中的表达变化。方法:采用Western blot和免疫组织化学荧光法检测脑损伤后CRM1在脑中的表达变化。结果:(1)假手术组皮质CRM1蛋白水平低,在损伤后12小时逐步增加,在损伤后3天达到高峰,之后恢复到正常水平。非手术对侧皮层中CRM1表达没有明显改变。(2)免疫组化荧光测定:损伤后CRM1表达水平不但上升,且有入核的亚定位变化,主要定位在神经元中。而假手术组和对侧则主要定位在胞浆中。结论:推测CRM1在创伤性脑损伤后与神经元凋亡有关。
基金Supported by National Natural Science Foundation of China,No.81441123(to Huang CM),No.31640053(to Lin Y)National Key Clinical Specialty Discipline Construction Program of China,No.[2012]649+1 种基金Key Scientific and Technological Project of Fujian Province,China,No.2014Y0025(to Huang CM)Natural Science Foundation of Fujian Province,China,No.2014J01322(to Xie JW),No.2016Y0029(to Lin Y)
文摘AIM To evaluate the predictive value of the expression of chromosomal maintenance(CRM)1 and cyclindependent kinase(CDK)5 in gastric cancer(GC) patients after gastrectomy.METHODS A total of 240 GC patients who received standard gastrectomy were enrolled in the study. The expression level of CRM1 and CDK5 was detected by immunohistochemistry. The correlations between CRM1 and CDK5 expression and clinicopathological factors were explored. Univariate and multivariate survival analyses were used to identify prognostic factors for GC. Receiver operating characteristic analysis was used to compare the accuracy of the prediction of clinical outcome by the parameters. RESULTS The expression of CRM1 was significantly related to size of primary tumor(P = 0.005), Borrmann type(P = 0.006), degree of differentiation(P = 0.004), depth of invasion(P = 0.008), lymph node metastasis(P = 0.013), TNM stage(P = 0.002) and distant metastasis(P = 0.015). The expression of CDK5 was significantly related to sex(P = 0.048) and Lauren's classification(P = 0.011). Multivariate Cox regression analysis identified that CRM1 and CDK5 co-expression status was an independent prognostic factor for overall survival(OS) of patients with GC. Integration of CRM1 and CDK5 expression could provide additional prognostic value for OS compared with CRM1 or CDK5 expression alone(P = 0.001).CONCLUSION CRM1 and CDK5 co-expression was an independent prognostic factors for GC. Combined CRM1 and CDK5 expression could provide a prognostic model for OS of GC.
文摘目的:研究NF-κBp50、JAB1和CRM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及其意义。方法:采用免疫组化方法检测94例DLBCL中NF-κBp50、JAB1和CRM1的表达。结果:NF-κBp50、JAB1和CRM1在94例DLBCL的阳性表达率分别为70.2%(66/94)、77.7%(73/94)和69.1%(65/94)。NF-κBp50在DL-BCL中非生发中心型(non-GCB)中表达明显高于生发中心型(GCB),差异有统计学意义(P<0.01),但JAB1、CRM1与DLBCL分型无关(P>0.05)。NF-κBp50、JAB1和CRM1与DLBCL患者性别、年龄、发病部位均无关(P>0.05)。DL-BCL中NF-κBp50与JAB1、NF-κBp50与CRM1的表达均呈正相关(rs=0.209,P<0.05;rs=0.270,P<0.01)。结论:NF-κp50在DLBCL不同亚型之间表达的差异,可能有助于DLBCL亚型的诊断和预后的判断。JAB1和CRM1对NF-κBp50表达具有正向调节作用。
基金We thank our colleagues at the UPV/EHU Dr.Sonia Bañuelos(Dept.of Biochemistry and Molecular Biology),Dr.Gorka Prieto(Dept.of Communication Engineering)and Dr.Asier Fullaondo(Dept.of Genetics,Physical Anthropology and Animal Physiology)for their help in preparing Figures 2 and 3.Supported by grants from the Spanish Government MINECO-FEDER(SAF2014-57743-R)the Basque Country Government(IT634-13)the University of the Basque Country(UFI11/20),as well as a fellowship from the Basque Country Government(to MS)。
文摘Cellular homeostasis crucially relies on the correct nucleocytoplasmic distribution of a vast number of proteins and RNA molecules,which are shuttled in and out of the nucleus by specialized transport receptors.The nuclear export receptor XPO1,also called CRM1,mediates the translocation of hundreds of proteins and several classes of RNA to the cytoplasm,and thus regulates critical signaling pathways and cellular functions.The normal function of XPO1 appears to be often disrupted in malignant cells due to gene mutations or,most commonly,aberrant overexpression.Due to its important physiological roles and its frequent alteration in human tumors,XPO1 is a promising target for cancer therapy.XPO1 inhibitors have undergone extensive testing as therapeutic agents in preclinical models of cancer,with promising results.One of these inhibitors,Selinexor,is currently being evaluated in multiple clinical trials of different types of solid tumors and hematological malignancies.Here,we review several key aspects of XPO1 function,as well as the mechanisms that may lead to its alteration in cancer,and provide an update on the status of XPO1 inhibitors being developed as drugs for cancer therapy,including the definitive results of the first clinical trials with Selinexor that have been recently published.
文摘Objectives: Multi-drug resistance (MDR) to chemotherapy remains a major obstacle to overcome in the successful treatment of patients with cancers. It was recently discovered that Leptomycin B (LMB) reduces the paclitaxel-induced MDR in CCRF-CEM/Taxol cells. However, the mechanism remains unclear. Here we sought to explore the mechanism of LMB to reduce the MDR induced by paclitaxel. Results: LMB has remarkable cytotoxic effects in both sensitive CCRF-CEM and resistant CCRF-CEM/Taxol cell lines. The paclitaxel-induced MDR was reduced by 0.013 μm of LMB. Lower concentration of LMB regulated cell cycle progress, in situ expressions of P-gp, MRP1, and LRP, expression of CRM1, and localization of P-gp and CRM1 in CCRF-CEM/taxol cells. Study Design: Cytotoxicity of LMB on cancerous cell lines was determined by MTT assay. Cell cycle progress and in situ expressions of P-gp, MRP1, and LRP were analyzed by flow cytometry. Expression of CRM1 in the cells was examined by Western blot. And co-localization between P-gp and CRM1 was determined by laser confocal microscopy. Conclusion: The paclitaxel-induced MDR of CCRFCEM/Taxol cells was reduced by lower concentration of LMB. The mechanisms might be related to decreasing in situ expression of drug transporter proteins, promoting cell cycle progress, and altering co-localization between P-gp and CRM1 in the resistant cells.
基金the Major State Basic Research Development Program of China(2010CB530105 and 2011CB504802)the National Natural Science Foundation of China(30900059,30870120 and 81000736)the Start-up Fund of the Hundred Talents Program of the Chinese Academy of Sciences(20071010-141)
文摘The function of the herpes simplex virus type 1 （HSV-1） UL4 protein is still elusive. Our objective is to investigate the subcellular transport mechanism of the UL4 protein. In this study, fluorescence microscopy was employed to investigate the subcellular localization of UL4 and characterize the transport mechanism in living cells. By constructing a series of deletion mutants fused with enhanced yellow fluorescent protein （EYFP）, the nuclear export signals （NES） of UL4 were for the first time mapped to amino acid residues 178 to 186. In addition, the N-terminal 19 amino acids are identified to be required for the granule-like cytoplasmic pattem of UL4. Furthermore, the UL4 protein was demonstrated to be exported to the cytoplasm through the NES in a chromosomal region maintenance 1 （CRM1）-dependent manner involving RanGTP hydrolysis
