Knockdown of nucleophosmin induces S-phase arrest in HepG2 cells

Wang, Qing Qing; Zhang, Zhi Yi; Xiao, Jian; Yi, Chunhui; Li, Lin Zi; Huang, Yan; Yun, Jing Ping

doi:10.5732/cjc.011.10362

Cited by 8 publications

(7 citation statements)

References 36 publications

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“…Analyzing only anagen VI HFs, quantitative immunohistomorphometry of the proliferation marker Ki-67 showed that, compared to scrambled control HFs, β1 integrin -specific silencing significantly reduced the number of Ki-67 + cells (10% less than scrambled control) in the maximally proliferating hair matrix ( Figure 1C ), and also significantly reduced the number of slow-cycling Ki-67 + cells in the HF bulge ( Figure 1D ). These results were double-checked by measuring EdU incorporation, a cell cycle S-phase specific marker to determine active DNA synthesis [ 46 ]. Counting EdU + cells in defined reference areas in the HF bulb and HF bulge, the same proliferation-inhibitory tendency after β1 integrin silencing could be demonstrated in both HF compartments ( Figure 1E , Figure S2C ).…”

Section: Resultsmentioning

confidence: 99%

“…A more specific method to detect cell proliferation is the quantifying of only S-phase active and DNA-synthesizing cells [ 45 ]. By using 5-ethynyl-2'-deoxyuridine (EdU), a terminal nucleoside analog of thymidine, its incorporation during active DNA synthesis [ 46 ] could be visualized because of its labeling with a stabile fluorescence dye. The whole method was performed following the manufacture’s guidelines (Click-iT® EdU Alexa Fluor® 488 Flow Cytometry Assay Kit, Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

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β1 Integrin Signaling Maintains Human Epithelial Progenitor Cell Survival In Situ and Controls Proliferation, Apoptosis and Migration of Their Progeny

et al. 2013

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β1 integrin regulates multiple epithelial cell functions by connecting cells with the extracellular matrix (ECM). While β1 integrin-mediated signaling in murine epithelial stem cells is well-studied, its role in human adult epithelial progenitor cells (ePCs) in situ remains to be defined. Using microdissected, organ-cultured human scalp hair follicles (HFs) as a clinically relevant model for studying human ePCs within their natural topobiological habitat, β1 integrin-mediated signaling in ePC biology was explored by β1 integrin siRNA silencing, specific β1 integrin-binding antibodies and pharmacological inhibition of integrin-linked kinase (ILK), a key component of the integrin-induced signaling cascade. β1 integrin knock down reduced keratin 15 (K15) expression as well as the proliferation of outer root sheath keratinocytes (ORSKs). Embedding of HF epithelium into an ECM rich in β1 integrin ligands that mimic the HF mesenchyme significantly enhanced proliferation and migration of ORSKs, while K15 and CD200 gene and protein expression were inhibited. Employing ECM-embedded β1 integrin-activating or -inhibiting antibodies allowed to identify functionally distinct human ePC subpopulations in different compartments of the HF epithelium. The β1 integrin-inhibitory antibody reduced β1 integrin expression in situ and selectively enhanced proliferation of bulge ePCs, while the β1 integrin-stimulating antibody decreased hair matrix keratinocyte apoptosis and enhanced transferrin receptor (CD71) immunoreactivity, a marker of transit amplifying cells, but did not affect bulge ePC proliferation. That the putative ILK inhibitor QLT0267 significantly reduced ORSK migration and proliferation and induced massive ORSK apoptosis suggests a key role for ILK in mediating the ß1 integrin effects. Taken together, these findings demonstrate that ePCs in human HFs require β1 integrin-mediated signaling for survival, adhesion, and migration, and that different human HF ePC subpopulations differ in their response to β1 integrin signaling. These insights may be exploited for cell-based regenerative medicine strategies that employ human HF-derived ePCs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

β1 Integrin Signaling Maintains Human Epithelial Progenitor Cell Survival In Situ and Controls Proliferation, Apoptosis and Migration of Their Progeny

et al. 2013

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“…Apoptosis is regulated by two major pathways: The death receptor-induced extrinsic pathway and the mitochondria-apoptosome-mediated intrinsic pathway (19). Bcl-2 family proteins are central in controlling the mitochondrial pathway, and >20 members of this family have been identified, including Bcl-2, which is one of the proteins that suppresses apoptosis (20).…”

Section: Discussionmentioning

confidence: 99%

Anti-proliferative and cytoskeleton-disruptive effects of icariin on HepG2 cells

Wang

Song

Ren

2015

Molecular Medicine Reports

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Several biological properties of icariin have been identified, including its anticancer effect. However, the potential mechanisms underlying the effect of icariin on HepG2 hepatocellular carcinoma cells remain to be elucidated. The aim of the present study was to examine the effects of icariin on the proliferation and cytoskeleton of HepG2 cells. A 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5 diphenyltetrazolium bromide assay was used to assess the antiproliferative effects of icariin and to determine the optimal concentration and treatment schedule of icariin on the HepG2 cells. Cell cycle analysis was performed using fluorescence activated cell sorting, the protein expression of B‑cell lymphoma (Bcl)‑2 was determined using immunohistochemical and western blot analyses, and F‑actin in the cells was examined using confocal microscopy. The chemotherapeutic drug, oxaliplatin, was used as a positive control. The results demonstrated that the optimal concentration of icarrin to produce an antiproliferative effect on HepG2 cells was 10‑5 mol/l, and the optimal treatment duration was 72 h. The icariin group had a significantly higher proportion of cells in the G0/G1 phase, compared with the control group, treated with high glucose Dulbecco's modified Eagles medium with 10% fetal bovine serum (P<0.05). The proportion of HepG2 cells in the S phase was significantly lower in the oxaliplatin (24.19%; P<0.05) and icariin (21.07%; P<0.01) groups, compared with the control group (28.62%). Icariin markedly decreased the expression of Bcl‑2, compared with the control (P<0.01), and disrupted the polymerization of F‑actin filaments in the HepG2 cells. Therefore, the present study demonstrated that, at an optimum concentration of 10‑5 mol/l, icariin inhibited the proliferation of the HepG2 cells, promoted apoptosis by decreasing the expression of Bcl‑2, and disrupted the actin cytoskeleton.

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“…Immunofluorescence staining For immunofluorescence analysis [28] , 2×10 5 cells were plated into each well of a 6-well plate with coverslips. The cells were incubated with 10 μg/mL Chol:MβCD in the presence or absence of 3 μmol/L ezetimibe for 72 h. After the treatment, the cells were washed with PBS.…”

Section: Quantitation Of Intracellular Cholesterol (Hplc and Enzymaticmentioning

confidence: 99%

Ezetimibe suppresses cholesterol accumulation in lipid-loaded vascular smooth muscle cells in vitro via MAPK signaling

Qin

Yang

et al. 2014

Acta Pharmacol Sin

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Aim: To investigate the mechanisms of anti-atherosclerotic action of ezetimibe in rat vascular smooth muscle cells (VSMCs) in vitro. Methods: VSMCs of SD rats were cultured in the presence of Chol:MβCD (10 μg/mL) for 72 h, and intracellular lipid droplets and cholesterol levels were evaluated using Oil Red O staining, HPLC and Enzymatic Fluorescence Assay, respectively. The expression of caveolin-1, sterol response element-binding protein-1 (SREBP-1) and ERK1/2 were analyzed using Western blot assays. Translocation of SREBP-1 and ERK1/2 was detected with immunofluorescence. Results: Treatment with Chol:MβCD dramatically increased the cellular levels of total cholesterol (TC), cholesterol ester (CE) and free cholesterol (FC) in VSMCs, which led to the formation of foam cells. Furthermore, Chol:MβCD treatment significantly decreased the expression of caveolin-1, and stimulated the expression and nuclear translocation of SREBP-1 in VSMCs. Co-treatment with ezetimibe (3 μmol/L) significantly decreased the cellular levels of TC, CE and FC, which was accompanied by elevation of caveolin-1 expression, and by a reduction of SREBP-1 expression and nuclear translocation. Co-treatment with ezetimibe dose-dependently decreased the expression of phosphor-ERK1/2 (p-ERK1/2) in VSMCs. The ERK1/2 inhibitor PD98059 (50 μmol/L) altered the cholesterol level and the expression of p-ERK1/2, SREBP-1 and caveolin-1 in the same manner as ezetimibe did. Conclusion: Ezetimibe suppresses cholesterol accumulation in rat VSMCs in vitro by regulating SREBP-1 and caveolin-1 expression, possibly via the MAPK signaling pathway.

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Knockdown of nucleophosmin induces S-phase arrest in HepG2 cells

Cited by 8 publications

References 36 publications

β1 Integrin Signaling Maintains Human Epithelial Progenitor Cell Survival In Situ and Controls Proliferation, Apoptosis and Migration of Their Progeny

β1 Integrin Signaling Maintains Human Epithelial Progenitor Cell Survival In Situ and Controls Proliferation, Apoptosis and Migration of Their Progeny

Anti-proliferative and cytoskeleton-disruptive effects of icariin on HepG2 cells

Ezetimibe suppresses cholesterol accumulation in lipid-loaded vascular smooth muscle cells in vitro via MAPK signaling

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