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1.
Smetana K Klamová H Mikulenková D Pluskalová M Hrkal Z 《European journal of histochemistry : EJH》2006,50(2):119-124
Human myeloblasts were studied in bone marrow of patients suffering from chronic phase of chronic myeloid leukaemia to provide more information on the nucleolar diameter in these early granulocytic progenitors. These cells are a convenient model for such study since the number of myeloblasts in diagnostic bone marrow smears of investigated patients is larger than in not-leukemic persons because of the increased granulopoiesis. The nucleolar diameter was measured in myeloblasts after various cytochemical procedures such as methods for visualisation of RNA, DNA and proteins of AgNORs using digitized images and image processing. The results clearly demonstrated that values of the nucleolar diameter depended on the procedures used for visualising nucleoli. It seems to be also clear that a close relationship exists between the diameter of nucleoli and their number since the larger the number of nucleoli per cell the smaller their mean size. However, one of multiple nucleoli present in the nucleus is usually significantly larger. Moreover, the possibility exists that the variability of nucleolar diameter of leukemic myeloblasts and thus the heterogeneity of these cells might depend on various stages of the cell cycle as supported by nucleolar measurements on aging leukemic myeloblasts (K 562 cells) in vitro. Since the staining density of small and large nucleoli did not differ substantially after staining for RNA, it seems to be likely that the nucleolar size is directly related to the total RNA content in myeloblasts. In addition, karyometry combined with RNA cytochemistry still appears to be an useful tool to study nucleoli at the single cell level. 相似文献
2.
To the intranucleolar translocation of AgNORs in leukemic early granulocytic and plasmacytic precursors 总被引:2,自引:2,他引:0
Smetana K Klamová H Pluskalová M Stöckbauer P Hrkal Z 《Histochemistry and cell biology》2006,125(1-2):165-170
Early leukemic granulocytic and plasmacytic precursors were studied in vitro and in vivo to provide an information on the
intranucleolar distribution of AgNORs (silver stained nucleolus organizer regions). In most of these cells AgNORs appeared
as clusters of silver stained particles distributed in the whole nucleolar body. On the other hand, in some leukemic early
granulocytic precursors, i.e., in myeloblasts and promyelocytes enlarged AgNORs were translocated in the nucleolar peripheral
part. In addition, the number of translocated AgNORs at the nucleolar periphery was significantly smaller. Such translocation
of a reduced number of AgNORs was easily produced by experimental aging, i.e., starving of cultured leukemic early granulocytic precursors (HL-60 and K562 cells) in vitro and seems to be reversible. Similar translocation of a reduced number of AgNORs was also produced by aging of leukemic plasmacytic precursors. Thus,
the translocation of the reduced number of AgNORs to the nucleolar periphery in some blastic leukemic hematopoietic cells
might be an useful marker of their aging at the single cell level. However, more studies in this direction are required in
the future. 相似文献
3.
Banfalvi G Klaisz M Ujvarosi K Trencsenyi G Rozsa D Nagy G 《Apoptosis : an international journal on programmed cell death》2007,12(12):2271-2283
Exponentially growing human erythroleukemia K562 cells were synchronized by centrifugal elutriation prior to and after Co60 γ-irradiation (4 Gy). Forward scatter flow cytometry used for size analysis revealed the increase of an early apoptotic cell
population ranging from lower (0.05 C-value) to higher DNA content (∼1 C) as the cells progressed through the S phase. The
increase in cellular DNA content expressed in C-values correlated with apoptotic chromatin changes manifested as many small
apoptotic bodies in early S phase and larger but less numerous disintegrated apoptotic bodies in late S phase. Most significant
changes after exposure to γ-irradiation took place in early S phase resulting in an increase of nuclear size by more than
50%. Cell fractions containing irradiated cells showed enhanced growth arrest at 2.4 C-value, which was accompanied by apoptosis.
Apoptotic cell cycle arrest near to the G1/G0 checkpoint and apoptotic changes indicate that the radiation resistance of K562 cells is related to the bypass of the early
stage of the p53 apoptotic pathway. Apoptotic changes in chromatin structure induced by γ-irradiation indicate that these
injury-specific changes can be identified and distinguished from chromatin changes induced by UV radiation or heavy metals. 相似文献
4.
羟基脲(Hydroxyurea)对细胞周期与珠蛋白基因表达的影响(简报) 总被引:7,自引:1,他引:7
Although the hydroxyurea (HU) has been extensively studied, little is known of its molecular mechanism in controlling the expression of human globin gene and in modulating the progression of cell-cycle in K 562 cell. In the present study, the effect of hydroxyurea on proliferative kinetics of K 562 cells was examined by monitoring the number of cells during a period of 8 day's cell culture. Our results showed that there was a dose related decrease in cell growth when K562 cells were incubated with HU. Moreover, cell-cycle analysis demonstrated that HU had profound effect on cell-cycle distribution. In the case of the induced K 562 cells, there was an increased accumulation of cells in S phase and a decreased fraction of cells in G 1 and G 2 + M phase. Furthermore, HU could induce the expression of human beta-globin gene in the induced K 562 cells. Our results indicate that HU has a potential to inhibit the proliferation of K 562 cells and to stimulate the terminal differentiation of this cell. 相似文献
5.
J R Roberts E K Rowinsky R C Donehower J Robertson D C Allison 《The journal of histochemistry and cytochemistry》1989,37(11):1659-1665
We used reliable and relatively inexpensive equipment to make sequential sets of measurements of antitubulin immunofluorescence, Feulgen staining, and autoradiography on the same cells. This was done to evaluate tubulin conformations, DNA content, and [3H]-thymidine incorporation in cell lines sensitive (HL60) and resistant (K562) to the novel anti-tubulin chemotherapeutic agent taxol. Numbers of cells with microtubule bundles have been found to correlate with sensitivity to taxol by clonogenic assay for several leukemic cell lines. We have found that cells with "asters" produced by taxol exposure are in mitosis and that cells with taxol-induced "bundles" are in G0/G1, S, and G2 phases. We further found that S-phase cells with microtubule bundles in both sensitive (HL60) and resistant (K562) cell lines were able to incorporate [3H]-thymidine after 4-hr exposure to taxol. As microtubule bundles and asters occur in cells of the same cell cycle phases in both lines, we conclude that the greater frequency of cells with microtubule bundles reported for sensitive cells after taxol treatment cannot result from drug exclusion nor from different effects of the drug on cell microtubules in these two leukemic lines. 相似文献
6.
The three-dimensional (3D) organization of rDNA-containing chromatin was studied in structurally well preserved, interphase nuclei ofPisum sativum root tips by in situ hybridization using a biotinylated cDNA probe to the 18, 5.8 and 25 S rDNA sequences. The probe was detected by immunofluorescence and optical section images recorded either by video imaging or by using a confocal laser scanning microscope. Detailed 3D reconstructions were made of 12 nucleoli by projection of confocal optical sections. The probe labelled four perinucleolar heterochromatin sites, one pair 1.0–2.1 µm in diameter and the other 0.5–1.0 µm diameter. It also labelled intranucleolar structures including 300–500 nm spots emanating from the perinucleolar sites into the body of the nucleolus. The intranucleolar labelled structures emanating from the perinucleolar sites lay in discrete domains. Medium power observations of 22 fields of cells (6–30 cells per field) were made by optical sectioning using a video camera and computer deblurring. The arrangement of the perinucleolar sites was modelled in each cell and the arrangements examined for nonrandomness. The sites tended to be spaced out around the nucleolar periphery approximating a regular tetrahedral arrangement as if to minimize clustering and the large sites appeared to lie in a plane perpendicular to the root axis. Cells with multiple nucleoli did not have any preferred distribution of sites between nucleoli. These observations are discussed in terms of current models of rDNA organization. 相似文献
7.
Smetana K Zápotocky M Starková J Trka J 《European journal of histochemistry : EJH》2008,52(3):143-148
The present study was designed to provide more information on nucleoli in apoptotic cells,which were represented in the present study by cultured leukemic myeloblasts (Kasumi-1 cells). The apoptotic process in these cells was produced by trichostatin A (TSA) that is a histone deacetylase inhibitor with strong cytostatic effects. The selected TSA concentration added to cultures facilitated to study apoptotic and not-apoptotic cells in one and the same specimen. The nucleolar diameter and density were determined using computer assisted measurement and densitometry in specimens stained for RNA. In comparison with not-apoptotic cells, in apoptotic cells, nucleolar mean diameter did not change significantly and nucleolar RNA density was also not apparently different. On the other hand, the cytoplasmic RNA density in apoptotic cells was markedly reduced. Thus it seemed to be possible that the transcribed RNA remained "frozen"within the nucleolus but its transport to the cytoplasm decreased or stopped. However, the possibility of the RNA degradation in the cytoplasm of apoptotic cells based on the present study cannot be eliminated. At this occasion it should be added that AgNORs reflecting nucleolar biosynthetic and cell proliferation activity in apoptotic cells decreased in number or disappeared. The presented results also indicated that large nucleoli intensely stained for RNA need not be necessarily related to the high nucleolar biosynthetic or cell proliferation activity and may be also present in apoptotic cells responding to the cytostatic treatment. 相似文献
8.
Shuhua Pan Jun Hu Tingting Zheng Xinyuan Liu Yong Ju Chuanlian Xu 《Cytotechnology》2015,67(5):821-829
Oleanolic acid (OA) derivatives exhibit numerous pleiotropic effects in many cancers. The present study aimed to investigate the molecular mechanisms of 5′-amino-oleana-2,12-dieno[3,2-d]pyrimidin-28-oic acid (compound 4) and oleana-2,12-dieno[2,3-d]isoxazol-28-oic acid (compound 5) inducing apoptosis in human leukemia K562 cell. We investigated the effects of the compounds on K562 cell growth, apoptosis and cell cycle. The compounds showed strong inhibitory effects on K562 cell viability in a dose-dependent manner determined by the 3-(4,5-dimethylthiazoyl)-2,5-diphenyltetrazolium bromide assay and significantly increased chromatin condensation and apoptotic bodies in K562 cells. Flow cytometry assay suggested that the compounds induced inhibition of K562 cell proliferation associated with G1 phase arrest. In addition, the compounds inhibited Akt1 recruiting to membrane in CHO cells which express Akt1-EGFP constitutively and down-regulated the expression of pAkt1 in K562 cell. These results suggested that the compounds can efficiently inhibit proliferation and induce apoptosis perhaps involved in inactivation of Akt1. The OA derivatives may be potential chemotherapeutic agents for the treatment of human cancer. 相似文献
9.
目的:通过,IPA诱导K562细胞分化过程中干预细胞铁代谢探讨白血病细胞铁与细胞分化的关系及对EGR1mRNA表达的影响。方法:应用体外细胞培养技术通过细胞形态,细胞化学染色观察细胞生长分化情况;用FCM、RT—PCR等技术检细胞周期、细胞表面分化抗原CD33、CD14及EGR1mRNA的表达。结果:在,IPA诱导K562细胞分化过程中铁剥夺可明显抑制K562细胞生长,并可阻止,IPA诱导K562细胞分化,使K562细胞停止在S期。铁剥夺可降低,TPA诱导K562细胞分化过程中EGR1mRNA的表达。讨论:铁剥夺明显抑制K562细胞生长、阻止TPA诱导K562细胞分化,故铁剥夺剂(DFO)可能作为一种辅助抗癌药用于白血病的化疗,但由于它能阻止白血病细胞的分化,故不宜用于白血病的诱导分化治疗。铁剥夺使K562细胞分化过程中E—GR1mRNA表达降低可能参与了阻止TPA诱导K562细胞的分化过程。 相似文献
10.
James M. Jamison Jacques Gilloteaux Laszlo Perlaky Marc Thiry Karel Smetana Deborah Neal Karen McGuire Jack L. Summers 《The journal of histochemistry and cytochemistry》2010,58(7):635-651
Ascorbate and menadione (Apatone) in a ratio of 100:1 kills tumor cells by autoschizis. In this study, vitamin-induced changes in nucleolar structure were evaluated as markers of autoschizis. Human bladder carcinoma (T24) cells were overlain with vitamins or with culture medium. Supernatants were removed at 1-hr intervals from 1 to 4 hr, and the cells were washed with PBS and prepared for assay. Apatone produced marked alterations in nucleolar structure including redistribution of nucleolar components, formation of ring-shaped nucleoli, condensation and increase of the proportion of perinucleolar chromatin, and the enlargement of nucleolar fibrillar centers. Immunogold labeling of the nucleolar rRNA revealed a granular localization in treated and sham-treated cells, and immunogold labeling of the rDNA revealed a shift from the fibrillar centers to the condensed perinucleolar chromatin. Fibrillarin staining shifted from the fibrillar centers and adjacent regions to a more homogeneous staining of the entire nucleolus and was consistent with the percentage of autoschizic cells detected by flow cytometry. Because autoschizis entails sequential reactivation of DNase I and DNase II, and because the fibrillarin redistribution following DNase I and Apatone treatment is identical, it appears that the nucleolar and fibrillarin changes are markers of autoschizis. (J Histochem Cytochem 58:635–651, 2010) 相似文献
11.
A comparative study was performed of the arrangement of different nucleolar components during differentiation of trophoblast cell populations in the junctional zone of placenta (glycogen cells and trophospongium) and in the secondary giant cells. Each cell type is characterized by specific interrelation of nucleolar components. Some glycogen cells show signs of segregation of nucleolar components: strands of nucleolar components with fibrillar centers (FCs) are displaced to the periphery of the nucleolus and contact with the perinucleolar chromatin. Large reticular nucleoli in trophospongium cells contain many FCs which are gathered into several "chains" by strands of dense fibrillar component. Such a "chain" has also been found in nucleoli of secondary giant cells, with greater number of FCs in each "chain". Relationship between the arrangement of nucleolar components and the level of cell differentiation is discussed. 相似文献
12.
Barbie DA Kudlow BA Frock R Zhao J Johnson BR Dyson N Harlow E Kennedy BK 《Molecular and cellular biology》2004,24(2):595-607
In primary mammalian cells, DNA replication initiates in a small number of perinucleolar, lamin A/C-associated foci. During S-phase progression in proliferating cells, replication foci distribute to hundreds of sites throughout the nucleus. In contrast, we find that the limited perinucleolar replication sites persist throughout S phase as cells prepare to exit the cell cycle in response to contact inhibition, serum starvation, or replicative senescence. Proteins known to be involved in DNA synthesis, such as PCNA and DNA polymerase delta, are concentrated in perinucleolar foci throughout S phase under these conditions. Moreover, chromosomal loci are redirected toward the nucleolus and overlap with the perinucleolar replication foci in cells poised to undergo cell cycle exit. These same loci remain in the periphery of the nucleus during replication under highly proliferative conditions. These results suggest that mammalian cells undergo a large-scale reorganization of chromatin during the rounds of DNA replication that precede cell cycle exit. 相似文献
13.
14.
Chromatin folding in the interphase nucleus is not known. We compared the pattern of chromatin condensation in Indian muntjac, Chinese hamster ovary, murine pre B, and K562 human erythroleukemia cells during the cell cycle. Fluorescent microscopy showed that chromosome condensation follows a general pathway. Synchronized cells were reversibly permeabilized and used to isolate interphase chromatin structures. Based on their structures two major categories of intermediates were distinguished: (1) decondensed chromatin and (2) condensed chromosomal forms. (1) Chromatin forms were found between the G1 and mid-S phase involving veil-like, supercoiled, fibrous, ribboned structures; (2) condensing chromosomal forms appeared in the late-S, G2, and M phase, including strings, chromatin bodies, elongated pre-chromosomes, pre-condensed chromosomes, and metaphase chromosomes. Results demonstrate that interphase chromosomes are clustered in domains; condensing interphase chromosomes are linearly arranged. Our results raise questions related to telomer sequences and to the chemical nature of chromosome connectivity. 相似文献
15.
Qinghuang Chen Ke Chen Guijie Guo Fang Li Chao Chen Song Wang Grzegorz Nalepa Shile Huang Ji-Long Chen 《PloS one》2014,9(10)
CDKN3 (cyclin-dependent kinase inhibitor 3), a dual specificity protein phosphatase, dephosphorylates cyclin-dependent kinases (CDKs) and thus functions as a key negative regulator of cell cycle progression. Deregulation or mutations of CDNK3 have been implicated in various cancers. However, the role of CDKN3 in Bcr-Abl-mediated chronic myelogenous leukemia (CML) remains unknown. Here we found that CDKN3 acts as a tumor suppressor in Bcr-Abl-mediated leukemogenesis. Overexpression of CDKN3 sensitized the K562 leukemic cells to imanitib-induced apoptosis and dramatically inhibited K562 xenografted tumor growth in nude mouse model. Ectopic expression of CDKN3 significantly reduced the efficiency of Bcr-Abl-mediated transformation of FDCP1 cells to growth factor independence. In contrast, depletion of CDKN3 expression conferred resistance to imatinib-induced apoptosis in the leukemic cells and accelerated the growth of xenograph leukemia in mice. In addition, we found that CDKN3 mutant (CDKN3-C140S) devoid of the phosphatase activity failed to affect the K562 leukemic cell survival and xenografted tumor growth, suggesting that the phosphatase of CDKN3 was required for its tumor suppressor function. Furthermore, we observed that overexpression of CDKN3 reduced the leukemic cell survival by dephosphorylating CDK2, thereby inhibiting CDK2-dependent XIAP expression. Moreover, overexpression of CDKN3 delayed G1/S transition in K562 leukemic cells. Our results highlight the importance of CDKN3 in Bcr-Abl-mediated leukemogenesis, and provide new insights into diagnostics and therapeutics of the leukemia. 相似文献
16.
红毛五加茎皮多糖对体外培养人白血病粒细胞生物学效应的研究 总被引:13,自引:0,他引:13
本文采用体外培养方法就红毛五加茎皮多糖对癌细胞生物学效应及其抗癌机制进行了初探。实验结果证明,红毛五加茎皮多糖具有良好的抗癌效果,能使体外培养的人白血病粒细胞形态明显改变,癌细胞生长受抑制,细胞生长曲线下降,细胞死亡率上升,流式细胞分光光度计测定分析,提示癌细胞合成DNA的前期受到抑制,阻断DNA的合成。 相似文献
17.
Proliferating cell nuclear antigen (PCNA) and human malignant tumor nucleolar antigens (HMTNA) in nucleoli of human hematological malignancies 总被引:7,自引:0,他引:7
Lymphoma (Lymphocytic non-Hodgkin's malignant lymphoma) and leukemic (chronic lymphocytic, acute and chronic myeloid, myelomonocytic leukemia) cells were studied by indirect immunofluorescence to evaluate the presence of proliferating cell nuclear antigen (PCNA) and human malignant tumor nuclear antigen (HMTNA) in their nucleoli. Most cells in lymph node smears of lymphocytic non-Hodgkin's malignant lymphoma (NHML) developed a bright nucleolar fluorescence with HMTNA antibodies. PCNA was detected in nucleoli of a limited number of cells which apparently represent the proliferating cell population in these lymphomas. Similarly, in the bone marrow smears of patients with chronic lymphocytic leukemia most cells possessed a nucleolar fluorescence for HMTNA and PCNA was present in nucleoli of a limited number of cells. In the bone marrow smears of patients with myeloid or myelomonocytic leukemias most blastic or monocytoid cells also developed a bright nucleolar fluorescence with HMTNA antibodies and PCNA was present only in a small percentage of these cells. Leukemic cells with PCNA in their nucleoli like thekhuntigen might represent a proliferating cell population in late G1-early S phase. 相似文献
18.
19.
Cells of the human erythroleukemic line K562 can be induced by manipulation of culture conditions to arrest within the G1 phase of the cell cycle, and subsequently to enter S phase synchronously after release from G1. Cell cultures subjected to serum deprivation and hydroxyurea (HU) treatment demonstrated less than 5% of the cells to be in S phase. Four hours after release from HU, 63% of the cells were in S phase, as detected by immunofluorescent staining. This protocol offers a method for synchronization of K562 cells at the G1/S border and a technique for detection of S-phase cells without the use of radioisotopes or flow cytometry instrumentation. 相似文献
20.