To explore the existence and distribution of nucleophosmin in the nuclear matrix and its co-localization with the other related gene products following HMBA treatment in the human hepatocarcinoma SMMC-7721 cells,the n...To explore the existence and distribution of nucleophosmin in the nuclear matrix and its co-localization with the other related gene products following HMBA treatment in the human hepatocarcinoma SMMC-7721 cells,the nuclear matrix of SMMC-7721 cells was extracted pre/post HMBA induced differentiation.2D PAGE proteomics analyses showed that nucleophosmin existed in the fractions of nuclear matrix proteins and was down-regulated after HMBA treatment with further confirmation by Western blot analysis.The immunofluorescence observation revealed that nucleophosmin located in the nuclear matrix,HMBA treatment altered its expression level and distribution profile.The co-localization of nucleophosmin with cancer-related genes and the products of oncogenes or tumor repression genes,including c-fos,c-myc,p53 and Rb,using laser scanning confocal microscopy,were evaluated,and substantial differences were observed following HMBA treatment.The results implies that nucleophosmin,as a nuclear matrix protein,the level of its expression and the colocalization with cancer-related gene products may play an important role during the differentiation of SMMC-7721 cell.展开更多
The behavior of UBF (upstream binding factor) and nucleophosmin in HeLa and HeLa-Bcl-2 cells during apoptosis induced by TNF-α, emetine, and their mixture was investigated. A pronounced apoptosis was achieved only in...The behavior of UBF (upstream binding factor) and nucleophosmin in HeLa and HeLa-Bcl-2 cells during apoptosis induced by TNF-α, emetine, and their mixture was investigated. A pronounced apoptosis was achieved only in HeLa cells treated with a mixture of the inducers. Immunoblotting analysis of UBF and nucleophosmin in samples containing different portions of cells with apoptotic nuclei was carried out. It showed that UBF was proteolytically cleaved giving a stable 76-kDa fragment. Increasing content of the fragment during apoptosis correlated with the level of cells containing apoptotic nuclei and with a decrease in the content of full-sized UBF. Determination of N- and C-terminal sequences of UBF and 76-kDa fragment allowed us not only to characterize UBF at the protein level, but also to describe the site of the apoptosis-specific proteolysis. Nucleophosmin did not undergo proteolytic cleavage during apoptosis and its content was unchanged even in a sample containing 100% of cells with apoptotic nuclei. However in cells reached terminal stages of apoptosis, the balance between mono- and oligomeric forms of nucleophosmin changed due to depletion of monomeric forms and appearance of two additional oligomeric forms with lower molecular weight.展开更多
Nucleophosmin/B23 (NPM) is a universally expressed nucleolar phosphoprotein that participates in proliferation, apoptosis, ribosome assembly, and centrosome duplication; however, the role of NPM in cell cycle regulati...Nucleophosmin/B23 (NPM) is a universally expressed nucleolar phosphoprotein that participates in proliferation, apoptosis, ribosome assembly, and centrosome duplication; however, the role of NPM in cell cycle regulation is not well characterized. We investigated the mechanism by which NPM is involved in cell cycle regulation. NPM was knocked down using siRNA in HepG2 hepatoblastoma cells. NPM translocation following actinomycin D (ActD) treatment was investigated using immunofluorescent staining. Expression of NPM and other factors involved in cell cycle regulation was examined by Western blotting. Cell cycle distribution was measured using flow cytometry to detect 5-ethynyl-2′-deoxyuridine (EdU) incorporation. Cell proliferation was quantified by the MTT assay. Knockdown of NPM increased the percentage of HepG2 cells in S phase and led to decreased expression of P53 and P21Cip1/WAF1. S-phase arrest in HepG2 cells was significantly enhanced by ActD treatment. Furthermore, knockdown of NPM abrogated ActD-induced G2/M phase cell cycle arrest. Taken together, these data demonstrate that inhibition of NPM has a significant effect on the cell cycle.展开更多
Background Nucleophosmin plays a critical role in embryonic development. This study aimed to examine the expression pattern of nucleophosmin in glandular epithelium of human endometrium during the menstrual cycle. Met...Background Nucleophosmin plays a critical role in embryonic development. This study aimed to examine the expression pattern of nucleophosmin in glandular epithelium of human endometrium during the menstrual cycle. Methods Endometrial tissues used for this study were obtained from 46 non-pregnant patients who underwent hysterectomy which had been performed to treat benign diseases. Nucleophosmin expression was assessed by in situ hybridization and immunohistochemistry. Results At the early-, mid- and late-proliferative phase, nucleophosmin mRNA was highly expressed in glandular epithelium of human endometrium. At the secretory phase, the expression of nucleophosmin mRNA was reduced in glandular epithelium in early-secretory phase, and the expression in mid- and late-secretory phases was not detected. Similarly, nucleophosmin protein was strongly expressed in endometrial glands throughout the proliferative phase, but was gradually reduced during secretory phase. Conclusion Nucleophosmin mRNA and protein are expressed in glandular epithelium of human endometrium throucIhout the menstrual cycle.展开更多
文摘To explore the existence and distribution of nucleophosmin in the nuclear matrix and its co-localization with the other related gene products following HMBA treatment in the human hepatocarcinoma SMMC-7721 cells,the nuclear matrix of SMMC-7721 cells was extracted pre/post HMBA induced differentiation.2D PAGE proteomics analyses showed that nucleophosmin existed in the fractions of nuclear matrix proteins and was down-regulated after HMBA treatment with further confirmation by Western blot analysis.The immunofluorescence observation revealed that nucleophosmin located in the nuclear matrix,HMBA treatment altered its expression level and distribution profile.The co-localization of nucleophosmin with cancer-related genes and the products of oncogenes or tumor repression genes,including c-fos,c-myc,p53 and Rb,using laser scanning confocal microscopy,were evaluated,and substantial differences were observed following HMBA treatment.The results implies that nucleophosmin,as a nuclear matrix protein,the level of its expression and the colocalization with cancer-related gene products may play an important role during the differentiation of SMMC-7721 cell.
文摘The behavior of UBF (upstream binding factor) and nucleophosmin in HeLa and HeLa-Bcl-2 cells during apoptosis induced by TNF-α, emetine, and their mixture was investigated. A pronounced apoptosis was achieved only in HeLa cells treated with a mixture of the inducers. Immunoblotting analysis of UBF and nucleophosmin in samples containing different portions of cells with apoptotic nuclei was carried out. It showed that UBF was proteolytically cleaved giving a stable 76-kDa fragment. Increasing content of the fragment during apoptosis correlated with the level of cells containing apoptotic nuclei and with a decrease in the content of full-sized UBF. Determination of N- and C-terminal sequences of UBF and 76-kDa fragment allowed us not only to characterize UBF at the protein level, but also to describe the site of the apoptosis-specific proteolysis. Nucleophosmin did not undergo proteolytic cleavage during apoptosis and its content was unchanged even in a sample containing 100% of cells with apoptotic nuclei. However in cells reached terminal stages of apoptosis, the balance between mono- and oligomeric forms of nucleophosmin changed due to depletion of monomeric forms and appearance of two additional oligomeric forms with lower molecular weight.
基金supported by grants from theNational Natural Science Foundation of China (No.81172345 and No. 30973506)the National High Technology Research and Development Program of China (863 Program) (2006AA02A404 to Y-X.Z.)
文摘Nucleophosmin/B23 (NPM) is a universally expressed nucleolar phosphoprotein that participates in proliferation, apoptosis, ribosome assembly, and centrosome duplication; however, the role of NPM in cell cycle regulation is not well characterized. We investigated the mechanism by which NPM is involved in cell cycle regulation. NPM was knocked down using siRNA in HepG2 hepatoblastoma cells. NPM translocation following actinomycin D (ActD) treatment was investigated using immunofluorescent staining. Expression of NPM and other factors involved in cell cycle regulation was examined by Western blotting. Cell cycle distribution was measured using flow cytometry to detect 5-ethynyl-2′-deoxyuridine (EdU) incorporation. Cell proliferation was quantified by the MTT assay. Knockdown of NPM increased the percentage of HepG2 cells in S phase and led to decreased expression of P53 and P21Cip1/WAF1. S-phase arrest in HepG2 cells was significantly enhanced by ActD treatment. Furthermore, knockdown of NPM abrogated ActD-induced G2/M phase cell cycle arrest. Taken together, these data demonstrate that inhibition of NPM has a significant effect on the cell cycle.
文摘目的探讨核仁磷酸蛋白1突变(nucleophosmin 1-mutated,NPM1m)的分型、变异等位基因频率(variant allele frequency,VAF)、共突变等对急性髓系白血病(acute myeloid leukemia,AML)患者预后的影响。方法回顾性分析2017年6月至2024年9月就诊于兰州大学第一医院血液科86例NPM1m-AML患者的临床资料及基因突变信息与生存情况,相关性分析采用Spearman相关系数,生存分析采用Kaplan-Meier曲线和Log-rank检验,使用Cox回归分析探究生存影响因素。结果NPM1m分型为罕见分型(Rares型)的患者总生存期(overall survival,OS)较ABD型缩短(中位OS:164天vs416天,P=0.043)。NPM1m VAF值与初诊外周血白细胞计数、乳酸脱氢酶(lactate dehydrogenase,LDH)含量呈正相关(P<0.05),VAF≥0.37时OS缩短(中位OS:164天vs 730天,P=0.003)。合并骨髓增生异常相关基因(myelodysplasia-related gene,MR)突变较MR野生型预后差(中位OS:45天vs 395天,P<0.001)、FMS样酪氨酸激酶3内部串联重复(FMS-like tyrosine kinase 3-internal tandem duplication,FLT3-ITD)及DNMT3A三突变较非三突变预后差(中位OS:173 d vs 483 d,P=0.007)。合并PTPN11突变改善预后(中位OS:395 d vs 240 d,P=0.027),而合并N/KRAS突变较野生型患者呈现出预后更好的趋势(中位OS:662 d vs 189 d,P=0.070),但差异无统计学意义。多因素Cox分析显示LDH(HR=1.002,95%CI:1.000~1.003,P=0.005)、NPM1m VAF值(HR=2.415,95%CI:1.208~4.829,P=0.013)、Rares型(HR=3.037,95%CI:1.134~8.136,P=0.027)、MR突变(HR=5.283,95%CI:1.991~14.017,P<0.001)为患者OS相关的独立危险因素。结论NPM1m-AML的预后分层需结合NPM1m的分子异质性,Rares型、VAF≥0.37、合并FLT3-ITD及DNMT3A突变、MR突变是不良预后因素,合并PTPN11突变改善患者预后,合并N/KRAS显示出预后更好的趋势。LDH、NPM1m VAF值、Rares型、MR突变是影响NPM1m-AML患者OS的独立危险因素。
文摘Background Nucleophosmin plays a critical role in embryonic development. This study aimed to examine the expression pattern of nucleophosmin in glandular epithelium of human endometrium during the menstrual cycle. Methods Endometrial tissues used for this study were obtained from 46 non-pregnant patients who underwent hysterectomy which had been performed to treat benign diseases. Nucleophosmin expression was assessed by in situ hybridization and immunohistochemistry. Results At the early-, mid- and late-proliferative phase, nucleophosmin mRNA was highly expressed in glandular epithelium of human endometrium. At the secretory phase, the expression of nucleophosmin mRNA was reduced in glandular epithelium in early-secretory phase, and the expression in mid- and late-secretory phases was not detected. Similarly, nucleophosmin protein was strongly expressed in endometrial glands throughout the proliferative phase, but was gradually reduced during secretory phase. Conclusion Nucleophosmin mRNA and protein are expressed in glandular epithelium of human endometrium throucIhout the menstrual cycle.
