Effects of estradiol on cell cycle and cyclin proteins of vascular smooth muscle cells in rats

To study the effects of 17β-estradiol(E2) on the growth of cultured rat vascular smooth muscle cells (VSMC).Methods The cell cycle and the expressions of Cyclin D1 and CDK4 proteins were examined by flow cytometry in VSMC cultured in different concentrations (0～100 nmol/L) of 17β-estradiol with or without serum.Results Under serum-stimulating conditions,17β-estradiol(1,10,100 nmol/L) promoted VSMC proliferation by accelerating their cell cycle progression from G1 to S phases,and the cell rates at S were (31.89±9.14)%(35.90±4.59)% and (30.77±1.20)% respectively,significantly higher than the corresponding values of control cells (21.63±1.80)%.This was accompanied by the significantly increased expression of Cyclin D1 and CDK4 proteins.In the cultures without serum,however,high concentrations (10,100 nmol/L) of E2 induced a cell cycle arrest at G1 phase,which was characterizsed by decreased cell rates at S phase [(9.93±1.43)% and (8.76±1.80)% respectively,P<0.05] as compared with the corresponding control values and a down-regulation of expressions of Cyclin D1 and CDK4 proteins.Conclusion E2 can either promote or inhibit VSMC proliferation depending upon the presence or absence of serum mitogens.The underlying mechanism may be associated with the hormone’s action on the expression of Cyclin D1 and CDK4 which act as the G1 phase regulators.4 refs.

Variation of Cyclin B1-like Protein During the Cell Cycle of Physarum polycephalum

Physarum polycephalum L., a naturally synchronized myxomycophyta, was demonstrated to contain a cyclin B1-like protein by Western blot and immunoelectron microscopy. The content and subcellular location of the protein varied during the cell cycle. The cyclin B1-like protein was first detected in the plasmodia of S phase while it did not appear in the nuclei until late G2 phase. The content of the protein in both the plasmodia and nuclei rose gradually onwards, peaked at metaphase and disappeared abruptly at ana-telophase. The protein was found to be distributed in both the cytoplasm and nuclei in late G2 phase and metaphase. In nuclei, the protein was mainly located in the chromosomal and nucleolar areas. The results suggest that the cyclin B1-like protein of P. polycephalum begins to be synthesized at S phase, enters the nuclei at late G2 phase, accumulates in both cytoplasm and nuclei onwards and breaks down at ana-telophase. The results also suggest that the cyclin B1-like protein acts as a cytoplasmic-nuclear protein during certain phases of the cell cycle.

子宫平滑肌肉瘤组织中Dicer蛋白、Cyclin-D1的表达及其对患者预后的预测价值

目的探究Dicer蛋白、细胞周期蛋白-D1(Cyclin-D1)在子宫平滑肌肉瘤组织中的表达及其对患者预后的预测价值。方法回顾性选取2015年7月至2021年7月期间收治并完成随访的68例子宫平滑肌肉瘤患者作为研究对象,取患者手术切除瘤组织和瘤旁正常组织进行分组。比较两组组织和不同子宫平滑肌肉瘤患者Dicer蛋白、Cyclin-D1表达的差异,采用KM生存分析不同Dicer蛋白、Cyclin-D1表达患者的生存情况,采用COX回归分析影响子宫平滑肌肉瘤患者预后的危险因素,采用受试者工作特征(ROC)曲线分析Dicer蛋白、Cyclin-D1表达预测子宫平滑肌肉瘤预后的价值。结果子宫平滑肌肉瘤中Dicer蛋白阳性和Cyclin-D1阳性表达分别为70.59%(48/68)、58.82%(40/68),明显高于瘤旁组织的10.29%(7/68)、8.82%(6/68),差异均有统计学意义(P<0.05);子宫平滑肌肉瘤组织中Dicer蛋白阳性表达、Cyclin-D1阳性表达均与年龄无相关性(P>0.05),但均与TNM分期、淋巴结受累存在相关性(P<0.05)。68例患者死亡22例(32.35%);Kaplan-Meier生存分析结果显示,Dicer蛋白阴性表达组生存状况优于Dicer蛋白阳性表达组,Cyclin-D1阴性表达组生存状况优于Cyclin-D1阳性表达组,差异均有统计学意义(P<0.05)。COX回归分析结果显示,Dicer蛋白阳性、Cyclin-D1阳性、TNM分期(Ⅲ期)、淋巴结受累均为子宫平滑肌肉瘤患者预后的危险因素(P<0.05);ROC曲线分析结果显示,Dicer蛋白、Cyclin-D1表达预测子宫平滑肌肉瘤预后的AUC分别为0.650、0.636,均具有一定准确性。结论Dicer蛋白、Cyclin-D1在子宫平滑肌肉瘤组织中高表达,与患者临床病理特征与预后状况密切相关,可作为预测子宫平滑肌肉瘤预后有效指标。

Bcl-1 Rearrangement and Cyclin D1 Protein Expression in Multiple Myeloma Precursor Cells

The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL-1 rearrangement and cyclin D1 protein expression in 15 cases of MM were detected. By using hemi-nested polymerase chain reaction (PCR) the genomic DNA from fresh peripheral blood and bone marrow was amplified and the expression of cyclin D1 in the smears was detected by using immunohistochemical method. Ten volunteer with normal bone marrow served as control group. The results showed Bcl-1 rearrangement was detectable in 3/15 (20 %) MM patients and cyclin D, expression in 4/15 (27 % ) MM patients. BeLl-1 rearrangement and cyclin D1 protein expression were also detected in MM precursor cells. No overexpression of cyclin D1 or the rearrangement of the BeL-1 gene was found in the 10 volunteers. It was concluded that Bel-1 rearrangement and cyclin D1 protein overexpression were detected in MM precursor cells, speculating that overexpression of cyclin D1 protein may play an initial (critical) role in the pathogenesis of MM.

Expression of cyclin-dependent protein kinase 5 in the hippocampus of vascular dementia mice after cerebral ischemia and reperfusion

BACKGROUND： The p25-activated cyclin-dependent protein kinase 5 （Cdk5） may induce neuronal cell death and cause the development of dementia following cerebral ischemia and reperfusion. OBJECTIVE： To observe changes in the expression of Cdk5 and p25 in hippocampal tissue of vascular dementia mice at different time points following cerebral ischemia and reperfusion. DESIGN, TIME AND SETTING： A randomized, controlled animal experiment was performed in the clinical trial center of Hebei Provincial People＇s Hospital between September 2007 and October 2008. MATERIALS： Cdk5 rabbit anti-mouse polyclonal antibody, p35 rabbit anti-mouse polyclonal antibody, and β-actin mouse monoclonal antibody were purchased from Santa Cruz Biotechnology, Inc., USA; horseradish peroxidase-labeled goat anti-rabbit IgG and horseradish peroxidase-labeled goat anti-mice IgG were offered by Beijing Zhongshan Geldenbridye Biotechnology Co.,Ltd., China; the protein quantitative kit was produced by Applygen Gene Technology Corp., Beijing, China; cDNA reverse transcription and PCR amplification reagents were products of TianGen＆ Biotech （Beijing） Co.,Ltd., China. METHODS： One hundred and sixty male Kunming mice were randomly divided into two groups： a sham-operated group （n = 65） and a model group （n = 95）. Vascular dementia was induced with three periods of transient ischemia and reperfusion of the bilateral common carotid arteries. In the sham-operated group, the bilateral common carotid arteries were not blocked. MAIN OUTCOME MEASURES： Behavioral tests were done at four and six weeks post surgery. Pathological changes in the hippocampal CA1 region were observed with hematoxylin-eosin staining Cdk5 mRNA expression was examined by RT-PCR, and Western blots were used to evaluate Cdk5 and p25 expression. Learning and memory performance were assayed using the Morris water maze. RESULTS： Vascular dementia reduced learning and memory performance at 4 and 6 weeks post surgery. Vascular dementia also caused severe, time-dependent neuronal damage and death in the hippocampal CA1 region. Dementia induction also increased mRNA and protein expression of Cdk5 and p25 at both 4 and 6 weeks after surgery. CONCLUSION： Cdk5/p25 is involved in the development of vascular dementia in mice following cerebral ischemia and reperfusion.

Expressions of beta-catenin, APC Protein, C-myc and Cyclin D1 in Ovarian Epithelial Tumor and Their Implication

Objective： To investigate the expressions of beta-catenin, protein APC （adenomatous polyposis coil protein）, c-myc and cyclin D1 and their implication in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expressions of beta-catenin, APC protein, c-myc and cyclin D1 in ovarian epithelial tumor in 48 cases. Results： The abnormal expression rate of beta-catenin in malignant and borderline ovarian epithelial tumors was higher than that in benign epithelial tumors （P〈0.01）. The expression rates of c-myc and cyclin-D1 in ovarian malignant and borderline epithelial tumors were higher than those in benign epithelial tumors too（P〈0.05）. The prevalence of APC protein positive expression in benign epithelial tumors were significantly greater than that in malignant epithelial tumors （P〈0.05）. A significant negative correlation was found between beta-catenin and APC protein in ovarian epithelial tumors; while a significant positive correlation was found between beta-catenin, c-myc and cyclin-D1 in ovarian epithelial tumor （P〈0.05）. Conclusion： The abnormal expressions of Beta-catenin, APC protein, c-myc and cyclin-D1 might be used to indicate the malignance transform of ovarian epithelial tumors.

EXPRESSION OF p16,CYCLIN D1 AND RB PROTEIN IN GASTRIC CARCINOMA AND PREMALIGNANT LESIONS

Objective: To investigate the expression of p16, cyclin D1 and Rb protein in gastric carcinoma and premalignant lesions including dysplastic gastric mucosa and intestinal metaplasia gastric mucosa. Methods: Using SP immunohistochemical methods, the expression of pl6, cyclin D1 and Rb proteins was detected in 10 specimens of normal gastric mucosa, 15 specimens of dysplastic gastric mucosa, 15 specimens of intestinal metaplasia gastric mucosa, 30 specimens of gastric carcinoma. The clinical characteristics of the 30 patients with gastric carcinoma were analysed to explore the relationship between the parameter detected and biological action of gastric cancer. Results: Expression of p16 protein was detected in 90% of normal gastric mucosa, 86.67% of dysplastic gastric mucosa, 86.67% of intestinal metaplasia gastric mucosa, 36.67% of gastric carcinoma. The positive rate of p16 protein expression in gastric carcinoma is significantly lower than that in normal gastric mucosa and gastric premalignant lesions mucosa (P<0.01). Expression of cyclin D1 protein was detected in 10% of normal gastric mucosa, 20% of dysplastic gastric mucosa, 20% of intestinal metaplasia gastric mucosa, 53.33% of gastric carcinoma. The positive rate of cyclin D1, protein expression in gastric carcinoma is significantly higher than that in normal gastric mucosa and gastric premalignant lesions mucosa (P<0.05). Expression of Rb protein was detected in 90% of normal gastric mucosa, 80% of dysplastic gastric mucosa, 80% of intestinal metaplasia gastric mucosa, 50% of gastric carcinoma. The positive rate of Rb protein expression in gastric carcinoma is significantly lower than that in normal gastric mucosa (P<0.05). The expression of p16, cyclin D1 gene were associated with the degree of differentiation of gastric carcinoma, lymphnodes metastasis and distant metastasis. Conclusion: p16, Cyclin D1 and Rb gene play important role in gastric carcinoma genesis. The expression of p16, cyclin D1 and Rb gene have some value to the diagnosis at earlier stage of gastric cancer. Detection of expression of p16, cyclin D1 gene would be helpful to judge the prognosis of gastric cancer.

人乳头瘤病毒16/18感染与膀胱癌组织cyclinD1、P53蛋白表达的相关性分析

目的 分析人乳头瘤病毒(HPV)16/18感染与膀胱癌组织细胞周期蛋白D1(cyclinD1)、P53蛋白表达的相关性。方法 采集62例膀胱癌患者的癌组织和癌旁组织。采用免疫组化染色和半定量积分方法,检测HPV16/18、cyclinD1和P53蛋白在膀胱癌及癌旁组织中的表达水平,并分析其表达与膀胱癌临床病理特征的关系以及THPV16/18与cyclinD1、P53蛋白表达的相关性。结果 膀胱癌组织中,HPV16/18、cyclinD1和P53蛋白的阳性率高于癌旁组织(P<0.05);HPV16/18、cyclinD1和P53蛋白阳性率病理分级高、临床分级高、有远处转移的膀胱癌组织分别高于病理分级低、临床分级低、无远处转移的膀胱癌组织(P<0.05);HPV16/18阳性组cyclinD1和P53蛋白阳性率高于HPV16/18阴性组(P<0.05);膀胱癌组织中HPV16/18表达与cyclinD1表达无相关性(P>0.05),与P53蛋白表达呈正相关(P<0.05)。结论 HPV16/18、cyclinD1和P53蛋白均参与了膀胱癌的发生和发展,HPV16/18可以影响P53蛋白的表达,参与膀胱癌的进展。

Bcl-3、cyclin D1、P53、Ki-67蛋白在乳腺导管内增生性病变的表达及其意义

目的探讨B细胞淋巴瘤因子3(Bcl-3)、细胞周期蛋白D1(cyclin D1)、P53基因编码蛋白和增殖细胞核蛋白(Ki-67)在乳腺导管内增生性病变中的表达及应用价值。方法采用免疫组化检测乳腺普通型导管增生(UDH)、柱状上皮病变(CCL)、非典型导管增生(ADH)和导管原位癌(DCIS)组织中Bcl-3、cyclin D1、P53和Ki-67蛋白的表达。结果Bcl-3、cyclin D1、P53、Ki-67蛋白在UDH中均少量表达;在CCL、ADH和DCIS中均明显表达,且随着病变的级别升高,阳性率逐渐增高(P<0.05);Bcl-3、cyclin D1、P53和Ki-67蛋白在CCL、ADH和DCIS中着色强阳性(++~+++)的表达,差异有统计学意义(P<0.05)。结论Bcl-3、cyclin D1、P53和Ki-67蛋白的表达在乳腺导管内增生性病变的病理诊断、鉴别诊断及指导临床治疗方面有较高应用价值,可显著提高乳腺导管内增生性病变的诊断准确率。

Cyclin D_2和p16蛋白表达在心肌细胞增殖、分化中的变化

目的 :探讨cyclinD2 、p16蛋白表达与新生大鼠心肌细胞体外培养早期增殖、分化的关系及意义。方法 :培养出生后 1d大鼠心肌细胞 ,予生长曲线、流式细胞计数检测培养过程中增殖改变 ,透射电镜观察超微结构变化 ,免疫细胞化学结合图像分析检测心肌细胞内cyclinD2 、p16蛋白变化。结果 :①生长曲线、流式细胞计数显示 :心肌细胞在培养的前 3d内具有一定增殖能力 ,但呈下降趋势同时逐步分化 ;3d后增殖能力丧失。超微结构显示细胞内有大量肌丝和线粒体。②心肌细胞在培养的 5d中 ,cyclinD2 表达量在第 3、4、5d分别是第 1d的 89 99% (P <0 0 5 )、83 6 1% (P <0 0 5 )和 6 9 6 5 % (P <0 0 1) ;p16表达量在 2、3、4、5d分别是第 1d的 1 6 3倍、1 72倍、1 99倍和2 84倍 (均P <0 0 1)。结论 :新生大鼠心肌细胞出生后早期体外培养前 3d有一定增殖能力 ,但呈下降趋势同时逐步分化 ;cyclinD2 表达下调和p16表达上升可能参与心肌细胞早期分化。

P27、CyclinD1、CyclinE在原发性胆囊癌中的表达及临床意义

目的探讨P27、CyclinD1和CyclinE在原发性胆囊癌组织中的表达及其临床意义。方法采用免疫组化SP法检测P27、CyclinD1和CyclinE在胆囊癌组织中的表达状况,并与同期手术的胆囊良性肿瘤和胆囊炎性疾病组织作对照。结果①P27在胆囊炎性组织中的高表达阳性率为80%(16/20),在胆囊良性肿瘤组织中为75%(15/20),两者均明显高于其在胆囊癌中的表达[37.3%(28/75),P<0.01];CyclinD1在胆囊癌组织中表达阳性率为58.7%(44/75),显著高于在胆囊良性肿瘤[15%(3/20)]和胆囊炎性疾病[10%(2/20)]中的表达(P<0.01)。Cy-clinE在胆囊癌组织中表达阳性率为60%(45/75),显著高于在胆囊良性肿瘤[15%(3/20)]和胆囊炎性疾病[10%(2/20)]中的表达(P<0.01)。②P27、CyclinD1和CyclinE的表达与胆囊癌病理分级、临床分期有关(P<0.01)并与胆囊癌患者的预后明显相关(P<0.01)。③P27和CyclinD1表达之间呈负相关(r=-0.864,P<0.01),P27和CyclinE表达之间呈负相关(r=-0.685,P<0.01),CyclinD1和CyclinE表达之间呈正相关(r=0.724,P<0.01)。结论P27表达下降,CyclinD1和CyclinE阳性高表达导致细胞周期的调控异常,该机制可能参与了胆囊癌的发生和发展过程。

p53、CyclinD1及PCNA在非小细胞肺癌的表达

目的 研究非小细胞肺癌 (NSCLC)中p5 3、CyclinD1蛋白表达及与细胞增殖的关系。方法 应用免疫组织化学技术 (SP法 )检测 74例NSCLC中p5 3、CyclinD1蛋白和增殖细胞核抗原 (PCNA)的表达情况。结果 p5 3蛋白、CyclinD1蛋白在NSCLC中阳性表达率分别为 5 5 .41%和 37.84% ,均明显高于正常肺组织 (P1= 0 .0 0 31,P2 =0 .0 42 9)。p5 3蛋白表达与淋巴结转移有密切关系 (P =0 .0 2 2 2 )。p5 3蛋白、CyclinD1蛋白表达分别与PCNA指数有密切关系 (P1 =0 .0 0 10 ,P2 =0 .0 0 0 9)。p5 3蛋白与CyclinD1蛋白表达之间未见明显关系(P >0 .0 5 )。结论 p5 3蛋白和CyclinD1蛋白过表达参与了NSCLC的发生发展 ,有促进细胞增殖的作用。p5 3是诊断和评价NSCLC预后的重要参数。

p27^(KIP1)和CyclinE在骨肉瘤中表达的意义

目的:研究p27KIP1和CyclinE蛋白在骨肉瘤中的表达,探讨其在骨肉瘤发生、发展中的可能作用。方法:应用免疫组化S-P方法检测28例骨肉瘤石蜡标本中p27KIP1和CyclinE蛋白的表达情况。结果:p27KIP1蛋白在28例骨肉瘤中表达阳性率为42.9%,在对照组20例骨软骨瘤中为75.0%,骨肉瘤与骨软骨瘤相比有显著差异(P<0.05)。CyclinE蛋白在28例骨肉瘤中表达阳性率为67.9%,在20例骨软骨瘤中为10.0%,骨肉瘤与骨软骨瘤相比有显著差异(P<0.01)。p27KIP1蛋白在骨肉瘤中的表达阴性率为57.1%,与CyclinE蛋白在骨肉瘤中表达阴性率67.9%相比二者差异不显著(P>0.05)。结论:p27KIP1在骨肉瘤中表达降低,CyclinE在骨肉瘤中高表达,二者与骨肉瘤发生机制有关,且具有相关性。

p-MAPK、cyclinD1和p53蛋白在大肠癌中的表达及其相关性

目的 :检测 p MAPK、cyclinD1和 p5 3蛋白在大肠癌中的表达 ,并探讨其相关性。 方法 :免疫组织化学S P法。结果 :140例大肠癌中 p MAPK、cyclinD1和p5 3蛋白的阳性表达率分别为 92 9% (130 / 140 )、87 1% (12 2 / 140 )和 6 6 4% (93/ 140 ) ;5 0例相应癌旁肠粘膜中阳性表达率分别为 4 0 % (2 / 5 0 )、6 0 % (3/ 5 0 )和 2 0 % (1/ 5 0 )。三者阳性表达率和阳性强度均以大肠癌中为高 (P <0 0 1) ;大肠癌中 p MAPK与cyclinD1表达呈明显正相关 (P <0 0 1) ,而与 p5 3蛋白表达无明显相关性 (P >0 .0 5 )。结论 :MAPK活化可能是cylinD1激活的原因之一 ,并在大肠癌发生过程中可能起重要作用 ;而 p5 3基因突变在大肠癌发生中可能不是通过MAPK信号传递通路起作用的。

EGFR和Cyclin D1的联合表达预测胶质瘤的放疗敏感性

目的探讨表皮生长因子受体(EGFR)和Cyclin D1蛋白的表达与胶质瘤放疗敏感性的关系。方法采用免疫组织化学方法检测EGFR和Cyclin D1蛋白在53例不同放疗敏感性胶质瘤和10例正常脑组织中的表达。结果EGFR或Cyclin D1蛋白的表达与胶质瘤的放疗敏感性均无统计学意义(P>0.05)。EGFR和Cyclin D1蛋白的联合表达与胶质瘤的放疗敏感性具有统计学意义(P<0.05)。结论EGFR和Cyclin D1蛋白的联合表达可预测胶质瘤的放疗敏感性。

P16和CyclinE蛋白在食管鳞状细胞癌中的表达及意义

目的探讨P16和cyclinE蛋白在食管鳞状上皮、增生上皮和癌变上皮中表达状况及其与鳞状细胞癌发生、发展和转移的相关性。方法采用SP免疫组织化学方法,检测食管鳞癌72例,粘膜上皮增生21例,正常对照13例和淋巴结转移癌15例中P16和cyclinE基因蛋白的表达。结果P16在正常上皮组无阳性表达,增生组为23.81%,鳞癌组为38.89%;高分化鳞癌组阳性表达率(41.67%)显著高于低分化鳞癌组(27.27%)(P<0.05);有淋巴结转移组(16.67%)和无淋巴结转移组(42.85%)比较具有显著性差异(P<0.05);原发癌组阳性率显著高于淋巴结转移癌组(P<0.05)。cyclinE在正常上皮组无阳性表达,增生上皮组为33.3%,鳞癌组为51.4%;高分化鳞癌组阳性表达率(44.44%)显著低于低分化鳞癌组(81.81%)(P<0.05);淋巴结转移组cyclinE阳性表达率(79.17%)和淋巴结无转移组(48.57%)比较有显著性差异(P<0.05)。P16和cyclinE表达与肿瘤部位和浸润深度相关性不明显。结论在食管癌中P16和cyclinE基因蛋白的表达与病理分化程度有关;检测P16和cyclinE对食管癌早期诊断有指导作用。

大黄鱼cyclin B1和cdc2 cDNA序列特征及组织表达分析

成熟促进因子（MPF）是诱导细胞从G2期转入M期的关键因子，在配子成熟过程中具有重要作用．MPF由周期蛋白B（cyelin13，CB）和周期蛋白依赖性蛋白激酶（cDKl，由cdc2基因编码）2个亚基组成．本研究克隆了大黄鱼（Larim-ichthyscrocea）cyclinB1（Lc-cbl）和cdc2（Lc-cdc2）基因的eDNA序列，并分析了这2个基因mRNA的组织表达特征．为解析MPF在大黄鱼性腺发育和配子成熟的作用机理奠定基础．Lc-cbl基因全长cDNA1882bp，可编码397个氨基酸的蛋白；Lc-cdc2基因全长eDNA序列1151bp，可编码303个氨基酸的蛋白．基于Lc-cblcDNA序列推导的氨基酸序列与6种脊椎动物的CBl氨基酸序列有较高相似性（67％～84％），并具有周期蛋白盒、毁坏盒、以及蛋白酶K位点（RRxSK）等CB预期特征．基于Lc-cdc2的eDNA序列推导的氨基酸序列也与其他6种鱼类的CDKl氨酸酸序列有较高相似性（88％～97％），并具有丝氨酸／苏氨酸激酶催化结构域、ATP结合相关的保守序列（GxGxxGxV）、周期蛋白结合相关的PSTAIRE序列等CDKl的预期特征．可见，本研究克隆获得的2条序列是Lc-cbl和Lc-cdc2eDNA全长．实时荧光定量PCR结果显示，Lc-cbl和Lc-cdc2的2个基因mRNA表达具有相似的组织特异性，在性腺中tuRNA水平均远远高于其他组织，表明Lc-c6J和Lc-cdc2是大黄鱼性腺发育相关的重要基因．

鼻咽癌细胞中EB病毒编码的潜伏膜蛋白1活化cyclinD1的表达

为了探讨EB病毒编码的潜伏膜蛋白 1(EBV LMP1)促进细胞增殖 ,参与EBV相关疾病致瘤的分子机制 ,研究了LMP1在鼻咽癌细胞中调节cyclinD1表达 ,进而影响细胞周期行进及细胞恶性表型改变 ,并初步确定了LMP1发挥该功能的结构域 .利用已建株的Tet on LMP1 HNE2鼻咽癌细胞系 ,蛋白质印迹实验分析LMP1诱导cyclinD1蛋白质表达的表达动力学 ,包括时间效应及剂量效应 ;利用三种LMP1功能区缺失的突变体及野生型LMP1,以载体型细胞为对照 ,确定LMP1活化cyclinD1表达的结构域 .同时结合基因诱导表达及反义寡聚核酸技术阻断基因表达的实验方法 ,进一步确定LMP1上调的cyclinD1功能 ,即对细胞周期行进及细胞恶性表型的影响 .结果表明LMP1确实可以诱导cyclinD1的表达 (2～ 4倍 ) ,且诱导具有时间依赖性及剂量依赖性 ;利用三种LMP1功能区缺失的突变体及野生型LMP1,以载体型细胞为对照 ,结合报道基因分析法 ,确定与空白载体细胞系比较 ,野生型LMP1从转录水平可反式激活cyclinD1报道基因活性约 11 2倍 ,其中CTAR1及CTAR2均可活化cyclinD1表达 ,但以CTAR2为主 ,与野生型LMP1诱导cyclinD1反式激活活性比较 ,CTAR1缺失导致cyclinD1报道基因活性下降 2 3 6 % ,CTAR2缺失导致cyclinD1活性下降约 80 7% ,C端均缺失时cyclinD1活性只?

乳腺癌组织中CyclinD1及p16蛋白表达的联合检测及其意义

目的:探讨乳腺癌中Cyclin D1及p16蛋白表达的相关性及其临床意义。方法:LSAB法检测多种乳腺组织中Cyclin D1及p16蛋白的表达情况,结合有关临床资料,分析Cyclin D1和p16蛋白表达异常与乳腺癌中重要的临床病理因素的关系。结果:62例乳腺癌组织中,Cyclin D1蛋白过度表达占48·4%,其表达与乳腺癌组织学分级呈明显正相关,且Cyclin D1蛋白过度表达更常见于ER、PR阳性的乳腺癌中。乳腺癌旁组织中Cyclin D1蛋白过度表达为20·0%,其他几种乳腺组织很少有CyclinD1蛋白的过度表达。p16蛋白仅在58·1%的乳腺癌组织中有表达,且p16的表达与组织学分级程度有负相关关系。正常乳腺组织中未见p16蛋白的异常。结论:乳腺癌组织中存在着Cyclin D1蛋白的过度表达及p16蛋白的异常,二者对乳腺癌组织的增殖分化有一定的影响。

PI3K抑制剂LY294002对人晶状体上皮细胞E2F-1、cyclinE蛋白表达的影响

目的探讨PI3K抑制剂LY294002对人晶状体上皮细胞(LECs)增殖及E2F-1、细胞周期素E(cyclinE)蛋白表达的影响。方法人LECs常规培养,分为实验组和对照组,实验组加入PI3K抑制剂LY294002(25μmol/L),对照组加入同体积DMSO,两组分别培养36 h后,MTT比色法测定细胞增殖状况,Western blot法检测细胞中E2F-1及cyclinE蛋白表达。结果实验组细胞增殖受到明显抑制,与对照组比较差异有统计学意义(P<0.05),人LECs中E2F-1及cyclinE蛋白表达降低(P<0.05)。结论 PI3K抑制剂LY294002可抑制人LECs的增殖,可能与下调E2F-1及cyclinE蛋白的表达水平相关。