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Quantification of the cell-cycle inhibitors p27(Kip1) and p21(Cip1) in human atherectomy specimens: primary stenosis versus restenosis 总被引:3,自引:0,他引:3
Braun-Dullaeus RC Ziegler A Bohle RM Bauer E Hein S Tillmanns H Haberbosch W 《The Journal of laboratory and clinical medicine》2003,141(3):179-189
Proliferation, a key determinator of vascular proliferative diseases, is dependent on cyclin/cyclin-dependent kinase (CDK) complexes, which are controlled by cyclin-dependent kinase inhibitors (CKIs) such as p27(Kip1) and p21(Cip1). Both have prognostic significance in various human malignancies. We have determined the levels of p27(Kip1) and p21(Cip1) in human directional coronary atherectomy specimens of primary lesions (n = 15) and lesions of in-stent restenosis (n = 18) in comparison to those of other vascular regions and have correlated CKI levels with clinical data. Quantitative immunoblotting demonstrated low expression of p27(Kip1) in primary lesions (5.9 +/- 0.5 ng/mg protein) compared with that in aorta (14.9 +/- 0.9 ng/mg), internal mammary artery (16.7 +/- 1.1 ng/mg), and carotid artery thrombendarterectomy specimens (16.5 +/- 1.7 ng/mg). Similarly, p27(Kip1) levels in lesions of in-stent restenosis were found to be significantly reduced (6.3 +/- 1.1 ng/mg; mean time of restenosis development 367 +/- 61 days). p27(Kip1) levels did, however, not have prognostic significance for the development of restenosis, and expression levels of proliferating cell nuclear antigen and CDK2 were similar in all groups examined, indicating low proliferative activity. Clinically, p27(Kip1) was not of value in predicting the development of restenosis. Furthermore, p27(Kip1) tissue levels were not increased in statin-treated patients, implying that the favorable effect of these drugs is not a result of p27(Kip1) stabilization. However, the relative content of p21(Cip1) was found to be significantly up-regulated in restenosis compared with that in primary lesions (225%) and the other vascular regions. Our data imply that negative-feedback mechanisms are still intact in coronary proliferative disease, thereby contrasting the finding of deregulated proliferation in malignancies. 相似文献
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Wiseman DA Werner SR Crowell PL 《The Journal of pharmacology and experimental therapeutics》2007,320(3):1163-1170
Pancreatic cancer, the fourth leading cause of cancer-associated mortality in the United States, usually presents in an advanced stage and is generally refractory to chemotherapy. As such, there is a great need for novel therapies for this disease. The naturally derived isoprenoids perillyl alcohol, farnesol, and geraniol have chemotherapeutic potential in pancreatic and other tumor types. However, their mechanisms of action in these systems are not completely defined. In this study, we investigated isoprenoid effects on the cell cycle and observed a similar antiproliferative mechanism of action among the three compounds. First, when given in combination, the isoprenoids exhibited an additive antiproliferative effect against MIA PaCa-2 human pancreatic cancer cells. Furthermore, all three compounds induced a G(0)/G(1) cell cycle arrest that coincided with an increase in the expression of the cyclin kinase inhibitor proteins p21(Cip1) and p27(Kip1) and a reduction in cyclin A, cyclin B1, and cyclin-dependent kinase (Cdk) 2 protein levels. Immunoprecipitation studies demonstrated increased association of both p21(Cip1) and p27(Kip1) with Cdk2 as well as diminished Cdk2 kinase activity after isoprenoid exposure, indicating a cell cycle-inhibitory role for p21(Cip1) and p27(Kip1) in pancreatic adenocarcinoma cells. When siRNA was used to inhibit expression of p21(Cip1) and p27(Kip1) proteins in MIA PaCa-2 cells, conditional resistance to all three isoprenoid compounds was evident. Given similar findings in this cell line and in BxPC-3 human pancreatic adenocarcinoma cells, we conclude that the chemotherapeutic isoprenoid compounds perillyl alcohol, farnesol, and geraniol invoke a p21(Cip1)- and p27(Kip1)-dependent antiproliferative mechanism in human pancreatic adenocarcinoma cells. 相似文献
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Modulation of apoptosis by the cyclin-dependent kinase inhibitor p27(Kip1) 总被引:13,自引:0,他引:13
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Hiromura K Pippin JW Fero ML Roberts JM Shankland SJ 《The Journal of clinical investigation》1999,103(5):597-604
Proliferation and apoptosis are increased in many types of inflammatory diseases. A role for the cyclin kinase inhibitor p27(Kip1) (p27) in limiting proliferation has been shown. In this study, we show that p27(-/-) mesangial cells and fibroblasts have strikingly elevated rates of apoptosis, not proliferation, when deprived of growth factors. Apoptosis was rescued by restoration of p27 expression. Cyclin A-cyclin-dependent kinase 2 (CDK2) activity, but not cyclin E-CDK2 activity, was increased in serum-starved p27(-/-) cells, and decreasing CDK2 activity, either pharmacologically (Roscovitine) or by a dominant-negative mutant, inhibited apoptosis. Our results show that a new biological function for the CDK inhibitor p27 is protection of cells from apoptosis by constraining CDK2 activity. These results suggest that CDK inhibitors are necessary for coordinating the cell cycle and cell-death programs so that cell viability is maintained during exit from the cell cycle. 相似文献
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DePinto W Chu XJ Yin X Smith M Packman K Goelzer P Lovey A Chen Y Qian H Hamid R Xiang Q Tovar C Blain R Nevins T Higgins B Luistro L Kolinsky K Felix B Hussain S Heimbrook D 《Molecular cancer therapeutics》2006,5(11):2644-2658
The cyclin-dependent protein kinases are key regulators of cell cycle progression. Aberrant expression or altered activity of distinct cyclin-dependent kinase (CDK) complexes results in escape of cells from cell cycle control, leading to unrestricted cell proliferation. CDK inhibitors have the potential to induce cell cycle arrest and apoptosis in cancer cells, and identifying small-molecule CDK inhibitors has been a major focus in cancer research. Several CDK inhibitors are entering the clinic, the most recent being selective CDK2 and CDK4 inhibitors. We have identified a diaminopyrimidine compound, R547, which is a potent and selective ATP-competitive CDK inhibitor. In cell-free assays, R547 effectively inhibited CDK1/cyclin B, CDK2/cyclin E, and CDK4/cyclin D1 (K(i) = 1-3 nmol/L) and was inactive (K(i) > 5,000 nmol/L) against a panel of >120 unrelated kinases. In vitro, R547 effectively inhibited the proliferation of tumor cell lines independent of multidrug resistant status, histologic type, retinoblastoma protein, or p53 status, with IC(50)s = 0.60 mumol/L. The growth-inhibitory activity is characterized by a cell cycle block at G(1) and G(2) phases and induction of apoptosis. R547 reduced phosphorylation of the cellular retinoblastoma protein at specific CDK phosphorylation sites at the same concentrations that induced cell cycle arrest, suggesting a potential pharmacodynamic marker for clinical use. In vivo, R547 showed antitumor activity in all of the models tested to date, including six human tumor xenografts and an orthotopic syngeneic rat model. R547 was efficacious with daily oral dosing as well as with once weekly i.v. dosing in established human tumor models and at the targeted efficacious exposures inhibited phosphorylation of the retinoblastoma protein in the tumors. The selective kinase inhibition profile and the preclinical antitumor activity of R547 suggest that it may be promising for development for use in the treatment of solid tumors. R547 is currently being evaluated in phase I clinical trials. 相似文献
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Upregulation of miR-370 contributes to the progression of gastric carcinoma via suppression of FOXO1
Conghai Fan Sheng Liu Yupeng Zhao Youqun Han Longqiu Yang Guorong Tao Qing Li Lei Zhang 《Biomedicine & Pharmacotherapy》2013
FOXO1 is downregulated in a number of cancers. However, the underlying mechanisms are poorly understood. In this study, we report that the expression of miR-370 was upregulated in gastric cancer cell lines and gastric cancer tissues. Overexpression of miR-370 in gastric cancer cells promoted the cell proliferation and anchorage-independent growth, while silencing of miR-370 showed opposite effects. miR-370-induced proliferation was correlated with the downregulation of cyclin-dependent kinase inhibitors, p27Kip1 and p21Cip1, and the upregulation of the cell cycle regulator cyclin D1. Furthermore, we identified that FOXO1 is the functional target of miR-370. Restored expression of FOXO1 together with miR-370 strongly abrogated miR-370-induced cell proliferation. Taken together, our results revealed a novel mechanism of FOXO1 suppression mediated by miR-370 in gastric cancer. 相似文献
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The CCNG2 gene that encodes the unconventional cyclin G2 was one of the few genes up-regulated on anti-human epidermal growth factor receptor 2 (HER2) antibody-mediated inhibition of HER2 signaling. The purpose of this study was to explore how HER2 signaling modulates cyclin G2 expression and the effect of elevated cyclin G2 on breast cancer cell growth. Treatment of breast cancer cells that overexpress HER2 (BT474, SKBr3, and MDAMB453) with the anti-HER2 antibody trastuzumab or its precursor 4D5 markedly up-regulated cyclin G2 mRNA in vitro and in vivo, as shown by real-time PCR. Immunoblot and immunofluorescence analysis with specific antibodies against cyclin G2 showed that anti-HER2 antibody significantly increased cyclin G2 protein expression and translocated the protein to the nucleus. Trastuzumab was not able to induce cyclin G2 expression in cells weakly expressing HER2 (MCF7) or in cells that had developed resistance to trastuzumab. Enforced expression of HER2 in T47D and MDAMB435 breast cancer cells reduced cyclin G2 levels. Collectively, these data suggest that HER2-mediated signaling negatively regulates cyclin G2 expression. Inhibition of phosphoinositide 3-kinase (LY294002), c-jun NH(2)-terminal kinase (SP600125), and mammalian target of rapamycin (mTOR)/p70 S6 kinase (p70S6K; rapamycin) increased cyclin G2 expression. In contrast, treatment with inhibitors of p38 mitogen-activated protein kinase (SB203580), mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2 (U0126), or phospholipase Cgamma (U73122) did not affect cyclin G2 expression. Anti-HER2 antibody in combination with LY294002, rapamycin, or SP600125 induced greater cyclin G2 expression than either agent alone. Ectopic expression of cyclin G2 inhibited cyclin-dependent kinase 2 activity, Rb phosphorylation, cell cycle progression, and cellular proliferation without affecting p27(Kip1) expression. Thus, cyclin G2 expression is modulated by HER2 signaling through multiple pathways including phosphoinositide 3-kinase, c-jun NH(2)-terminal kinase, and mTOR signaling. The negative effects of cyclin G2 on cell cycle and cell proliferation, which occur without altering p27(Kip1) levels, may contribute to the ability of trastuzumab to inhibit breast cancer cell growth. 相似文献
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Ann Packeu Tim Béghin Jean‐Paul De Backer Georges Vauquelin 《Fundamental & clinical pharmacology》2010,24(3):293-303
D2‐type dopamine receptors are major recognition sites for antipsychotic drugs. There are two splice variants: D2S and D2L with an additional 29 amino acid sequence in the third intracellular loop. Only little comparative information is hitherto available about their pharmacological properties and none of these studies dealt with intact cell systems. This prompted us to investigate the binding properties of [3H]‐raclopride, a hydrophilic benzamide, and [3H]‐spiperone, a highly hydrophobic butyrophenone, to intact CHO cells expressing recombinant human D2L‐receptors. Presently, we have repeated and extended this experimental approach to the human D2S‐receptors in the same cell system. Except for a slower dissociation of [3H]‐spiperone from D2S, the binding properties of these and other antagonists were not significantly different for both isoforms (P > 0.05). The very slow dissociation of the atypical antipsychotic clozapine was surprising in light of its low affinity. Two experiments pointed out the existence of non‐competitive interactions between raclopride and spiperone for D2S as well as D2L (A. Packeu, J. P. De Backer & G. Vauquelin, in preparation). Alongside the different physicochemical properties of these ligands, this finding fits with a model wherein the hydrophilic raclopride approaches the D2L‐receptor from the aqueous phase, while the hydrophobic spiperone approaches the receptor by lateral diffusion between the membrane lipids. These different modes of approach could imply the existence of topologically distinct ligand binding sites at D2‐receptors. 相似文献
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Takahashi T Yamasaki F Sudo T Itamochi H Adachi S Tamamori-Adachi M Ueno NT 《Molecular cancer therapeutics》2005,4(7):1039-1046
Cyclin A-associated kinases, such as cyclin-dependent kinase 2 (CDK2), participate in regulating cellular progression from G(1) to S to G(2), and CDK2 has also been implicated in the transition to mitosis. The antitumor properties of CDK inhibitors, alone or in combination with taxanes, are currently being examined in clinical trials. Here, we examined whether the activity of kinases associated with cyclin A (such as CDK2) is important in determining cellular sensitivity to paclitaxel, a taxane and mitotic inhibitor used in chemotherapy for breast and ovarian cancer. We used adenoviral suppression or overexpression to manipulate the expression of CDK2 and cyclin A in one breast cancer and three ovarian cancer cell lines with different sensitivities to paclitaxel and assessed protein expression, kinase activity, cell cycle distribution, and sensitivity to paclitaxel. Transfection of a dominant-negative (DN)-CDK2 evoked resistance to paclitaxel by preventing cellular progression to mitosis through loss of CDK1 activity. Reexpression of wild-type CDK2 in DN-CDK2-transfected cancer cells restored CDK2 activity but not paclitaxel sensitivity. However, expression of cyclin A in DN-CDK2-transfected cells restored their sensitivity to paclitaxel. Although CDK2 activity was not directly involved in paclitaxel sensitivity, cyclin A-associated kinases did up-regulate CDK1 via phosphorylation. We conclude that cyclin A-associated kinase activity is required for these cells to enter mitosis and undergo paclitaxel-induced cell death. Combining taxane chemotherapy with any drug targeting cyclin A-associated kinases (e.g., pure CDK2 inhibitors) should be done with caution, if at all, because of the potential for enhancing taxane resistance. 相似文献
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Sala SG Muñoz U Bartolomé F Bermejo F Martín-Requero A 《The Journal of pharmacology and experimental therapeutics》2008,324(1):352-359
Recent work has suggested that statins may exert beneficial effects on patients suffering from Alzheimer's disease (AD). The pharmacological effects of statins extend beyond their cholesterol-lowering properties. Based on the antineoplastic and apoptotic effects of statins in several cell types, we hypothesized that statins may be able to protect neurons by controlling the regulation of cell cycle. A growing body of evidence indicates that neurodegeneration involves the activation of cell cycle machinery in postmitotic neurons. We and others have presented direct evidence to support the hypothesis that the failure of cell cycle control is not restricted to neurons in AD patients, but that it occurs in peripheral cells as well. For these reasons, we found it worthy to study the role of simvastatin on cell proliferation in immortalized lymphocytes from AD patients. We report here that simvastatin (SIM) inhibits the serum-mediated enhancement of cell proliferation in AD by blocking the events critical for G(1)/S transition. SIM induces a partial blockade of retinoblastoma protein phosphorylation and inhibition of cyclin E/cyclin-dependent kinase (CDK)2 activity associated with increased levels of the CDK inhibitors p21(Cip1) and p27(kip1). These effects of SIM on AD lymphoblasts are dependent on inhibition of the proteasome-mediated degradation of p21 and p27 proteins. The antiproliferative effect of this natural statin may provide a therapeutic approach for AD disease. 相似文献
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E. Ioachim 《International journal of clinical practice》2008,62(11):1736-1743
Aberrations in the cell cycle regulators are common features of many tumours and several have been shown to have prognostic significant in colorectal cancer. The expression patterns of cyclins D1 and E as well as cyclin‐dependent kinase (CDK) inhibitors p21waf1/cip1 and p27kip1 and their interrelationship with other cell cycle checkpoint proteins [p53, pRb, Ki‐67 and proliferative cell nuclear antigen (PCNA)] were investigated in colorectal cancer in order to ascertain coregulation and influence on tumour behaviour or survival. These molecular markers were localisated immunohistochemically using the monoclonal antibodies anticyclin D1 (DCS‐6), anticyclin E (13A3), anti‐p21 (4D10), anti‐p27 (1B4), anti‐p53 (DO7), anti‐Rb (AB‐5), MIB1 and PC10 in colorectal cancer tissue from 97 patients. Data were analysed statistically using the spss software program. Overexpression of cyclin D1, cyclin E and p21waf1/cip1 proteins (>5% positive neoplastic cells) was observed in 5.9%, 30% and 7.2% of the cases respectively. Increased levels of cyclin D1 (p = 0.0001) and p21waf1/cip1 protein (p = 0.03) in tumours with mucous differentiation were observed. Overexpression of cyclin D1 was correlated with tumour stage (p = 0.03), the lymph node involvement (p = 0.02), as well as p21waf1/cip1 protein expression (p < 0.0001). Cyclin E was positively correlated with p21waf1/cip1 (p = 0.014), as well as with the cell proliferation as measured by PCNA‐labelling index (p = 0.011) and Ki‐67 score (p = 0.007). A positive relationship of p21waf1/cip1 expression with the proliferative‐associated index Ki‐67 was noted (p = 0.005). Downregulation of p27kip1 was observed in 47.4% of the cases and was correlated with downregulation of pRb (p = 0.002) and PCNA score (p = 0.004). The prognostic significance of cyclins D1, E and CDK inhibitors p21waf1/cip1, p27kip1 in determining the risk of recurrence and overall survival with both univariate (long‐rang test) and multivariate (Cox regression) methods of analysis showed no statistically significance differences. In conclusion, these findings suggest that, the levels of the cell cycle regulators studied, do not seems to have a prognostic value, in terms of predicting the risk of early recurrence and overall survival. In addition, the interrelationships, probably means their contribution to the regulation of cell growth, through different pathways in colorectal carcinogenesis. 相似文献
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The degradation of p27 and cancer 总被引:1,自引:0,他引:1
The cell cycle of eukaryotic cells is regulated by a series of protein complexes composed of cyclins and cyclin-dependent kinases (CDKs), the activity of which is suppressed by a group of CDK inhibitors (CKIs). Among the CKIs, p27 plays a pivotal role in the control of cell proliferation. Degradation of p27 is a critical event for reentry of cells into the cell cycle from G0 phase and occurs through ubiquitination by two ubiquitin ligase complexes (KPC and SCFSkP2) and subsequent degradation by the 26S-proteasome. A tumor suppressing function of p27 has been demonstrated in mouse models and studies of human tumors. This review will focus on the regulation of p27 proteolysis and its consequences for tumorigenesis. 相似文献
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Wang SW Pan SL Guh JH Chen HL Huang DM Chang YL Kuo SC Lee FY Teng CM 《The Journal of pharmacology and experimental therapeutics》2005,312(3):917-925
This study delineates the antiproliferative activities and in vivo efficacy of YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole] in human hepatocellular carcinoma cells. YC-1 inhibited the growth of HA22T and Hep3B cells in a concentration-dependent manner without significant cytotoxicity. YC-1 induced G(1) phase arrest in the cell cycle, as detected by an increase in the proportion of cells in the G(1) phase using FAC-Scan flow cytometric analysis. It was further shown that cGMP, p42/p44 mitogen-activated protein kinase, or AKT kinase-mediated signaling pathways did not contribute to the YC-1-induced effect. Of note, YC-1 induced a dramatic increase in the expression of cyclin-dependent kinase (CDK)-inhibitory protein, p21(CIP1/WAP1), and a modest increase in p27(KIP1). The association of p21(CIP1/WAP1) with CDK2 was markedly increased in cells responsive to YC-1. YC-1 did not modify the expression of cyclin D1, cyclin E, CDK2, or CDK4. In a corollary in vivo study, YC-1 induced dose-dependent inhibition of tumor growth in mice inoculated with HA22T cells. Immunohistochemical analysis revealed an inverse relationship between the staining of p21(CIP1/WAF) and the staining of Ki-67, a cell proliferation marker. Based on the results reported herein, we suggest that YC-1 induces cell cycle arrest and inhibits tumor growth both in vitro and in vivo via the up-regulation of p21(CIP1/WAP1) expression in HA22T cells. Because of this, YC-1 is a potential antitumor agent worthy of further investigation. 相似文献
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Cyclin D1 overexpression promotes cardiomyocyte DNA synthesis and multinucleation in transgenic mice. 总被引:6,自引:3,他引:6
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M H Soonpaa G Y Koh L Pajak S Jing H Wang M T Franklin K K Kim L J Field 《The Journal of clinical investigation》1997,99(11):2644-2654
D-type cyclin/cyclin-dependent kinase (CDK) complexes regulate transit through the restriction point of the cell cycle, and thus are required for the initiation of DNA synthesis. Transgenic mice which overexpress cyclin D1 in the heart were produced to determine if D-type cyclin deregulation would alter myocardial development. Cyclin D1 overexpression resulted in a concomitant increase in CDK4 levels in the adult myocardium, as well as modest increases in proliferating cell nuclear antigen and CDK2 levels. Flow cytometric and morphologic analyses of dispersed cell preparations indicated that the adult transgenic cardiomyocytes had abnormal patterns of multinucleation. Histochemical analyses confirmed a marked increase in number of cardiomyocyte nuclei in sections prepared from the transgenic mice as compared with those from control animals. Tritiated thymidine incorporation analyses revealed sustained cardiomyocyte DNA synthesis in adult transgenic hearts. 相似文献
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丁酸钠联合全反式维甲酸对MDS细胞株SKM-1的诱导分化作用及其机制初步探讨 总被引:2,自引:0,他引:2
本研究探讨丁酸钠(NaB)抑制MDS细胞株SKM-1细胞生长、诱导其分化的分子机制,并研究其与全反式维甲酸(ATRA)的协同作用。用台盼蓝拒染实验观察药物对细胞生长曲线的影响;四氮唑盐还原试验和细胞表面分化抗原检测观察药物对细胞的分化作用;流式细胞术分析细胞周期;RT-PCR检察D型细胞周期蛋白、CDK和P21在mRNA水平的表达。结果表明:NaB和(或)ATRA均可抑制SKM-1细胞的生长,诱导细胞分化,将细胞周期阻滞于G0/G1期;ATRA下调CDK6、CDK4、cyclin D3和cyclin D1 mRNA的水平;NaB下调CDK2、cyclin D2和cyclin D1 mRNA的水平;两药联用下调CDK6、CDK4、CDK2、cyclin D1、cyclin D2和cyclin D3 mRNA的水平;ATRA和(或)NaB均上调P21 mRNA的水平。结论:NaB诱导SKM-1的分化可能是通过上调P21 mRNA的水平和抑制cycLin D-CDK复合体的形成完成的,NaB与ATRA对SKM-1细胞株的分化有协同作用。 相似文献