p27~(Kip1)和CyclinD1在非小细胞肺癌中表达及意义

为了探讨p27Kip1蛋白和CyclinD1蛋白在非小细胞肺癌(NSCLC)中的表达及意义,收集临床手术切除的非小细胞肺癌组织蜡块64例及正常肺组织10例,应用免疫组化(S-P法)检测组织中p27Kip1蛋白和CyclinD1蛋白的表达,结合临床病理资料和随访资料进行回顾性研究。实验发现NSCLC组织中p27Kip1蛋白表达和CyclinD1蛋白表达均明显不同于正常肺组织(P<0.01)。p27Kip1蛋白表达降低与NSCLC肿瘤大小、病理分级、分期增加、淋巴结转移之间有相关性(P<0.05),但与肿瘤组织学分型无相关性(P>0.05)。CyclinD1蛋白过表达与组织学分型、肿瘤大小、病理分级、临床分期、淋巴结转移无相关性(P>0.05)。p27Kip1蛋白表达与CyclinD1蛋白表达之间呈显著负相关(P<0.01)。cox单因素及多因素分析,p27Kip1蛋白低表达及CyclinD1过表达是影响NSCLC患者预后的主要因素。实验结果显示,NSCLC组织中,p27Kip1蛋白表达降低,而CyclinD1过表达,二者与NSCLC的发生发展机制有关,可作为预后指标,有利于NSCLC患者预后判断及个体化治疗。     

p16和cyclinD1基因蛋白在卵巢囊腺瘤组织中的表达和意义

应用s-p免疫组织化学方法观察人卵巢囊腺癌,交界性囊腺瘤和良性囊腺瘤中细胞增殖周期调控因子p16和cy-clinD1的表达和意义。结果表明,在卵巢囊腺癌p16蛋白表达阳性率为45%。明显低于良性囊腺瘤（95%）,染色强度大部分为弱阳性或阳性（ ）,cyclinD1蛋白表达阳性率为80%,明显高于良性囊腺瘤（25%）,染色强度大部分为阳性（ ）,强阳性（ ）。p16蛋白表达阳性率随着卵巢囊腺瘤恶性程度的降低而升高。cyclinD1蛋白表达阳性率与卵巢囊腺瘤恶性程度有关,本研究结果表明p16蛋白抑制肿瘤细胞增殖,而cyclinD1蛋白促进肿瘤细胞增殖这一细胞增殖这一细胞增殖调控机制,可能与卵巢囊腺癌的发生发展有关。     

p27^kip1及相关分子skp2在肺癌组织中的表达及其临床意义

目的：研究细胞周期素依赖性激酶抑制蛋白27（p27^kip1）和细胞S相激酶相关蛋白2（skp2）在肺癌癌组织中的表达及意义。方法：选取于我院就诊的72例肺癌患者的肺癌组织和20例癌旁正常肺组织,采用免疫组化技术检测标本中p27^kip1和skp2的表达,并分析其与患者的临床病理之间的关系。结果：skp2在肺癌组织中的表达高于正常肺组织,而p27^kip1在肺癌组织中的表达低于正常肺组织,差异均有统计学意义（P〈0.05）,且两者的表达呈负相关关系,相关系数r=-0.855（P〈0.05）,skp2的表达与肺癌组织学类型、分化程度、TNM分期、淋巴结有无转移、吸烟与否及p27^kip1蛋白表达有关（P〈0.05）。结论：p27^kip1低表达和skp2高表达可能是肺癌发生发展的重要原因,可应用于临床诊治肺癌患者和判断预后。     

HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原的比较

目的由于检测SIV p27抗原试剂盒来源困难,有时不稳定,鉴于HIV-1 p24与SIVp27有较强的交叉抗原,本研究比较HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原得出的结果是否存在一定的相关性。方法 HIV-1 p24和SIV p27两种ELISA试剂盒定性和定量检测样品中SIV p27抗原,并对检测结果进行回归和相关分析。结果 HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原的灵敏度分别是150 pg/mL和62.5 pg/mL。两种试剂盒检测病毒液和血浆中SIV p27抗原的定性结果一致。定量结果的统计分析得出病毒液的直线回归决定系数R2=0.857,直线相关系数r=0.926,P〈0.01,直线正相关程度较高;血浆的直线回归决定系数R2=0.512,直线相关系数r=0.716,P〈0.05,直线正相关程度较低。结论 HIV-1 p24 ELISA试剂盒能够替代SIVp27 ELISA试剂盒定性检测病毒液和血浆中SIV p27抗原,但只能定量检测病毒液中SIV p27抗原。     

p27~(kip1)及相关分子skp2在肺癌组织中的表达及其临床意义

目的:研究细胞周期素依赖性激酶抑制蛋白27(p27kip1)和细胞S相激酶相关蛋白2(skp2)在肺癌癌组织中的表达及意义。方法:选取于我院就诊的72例肺癌患者的肺癌组织和20例癌旁正常肺组织,采用免疫组化技术检测标本中p27kip1和skp2的表达,并分析其与患者的临床病理之间的关系。结果:skp2在肺癌组织中的表达高于正常肺组织,而p27kip1在肺癌组织中的表达低于正常肺组织,差异均有统计学意义(P0.05),且两者的表达呈负相关关系,相关系数r=-0.855(P0.05),skp2的表达与肺癌组织学类型、分化程度、TNM分期、淋巴结有无转移、吸烟与否及p27kip1蛋白表达有关(P0.05)。结论:p27kip1低表达和skp2高表达可能是肺癌发生发展的重要原因,可应用于临床诊治肺癌患者和判断预后。     

p27kip1、Cyclin D1在卵巢癌中的表达及临床意义

目的探讨p27kip1、Cyclin D1在卵巢癌发生方面的意义.方法应用免疫组织化学方法及半定量分析方法,检测50例卵巢癌、19例卵巢良性上皮肿瘤、 13例正常卵巢组织中的p27kip1和Cyclin D1表达,并分析它们与良恶性肿瘤、病理学分级、临床分期的相关性. 结果正常卵巢和卵巢良性肿瘤间,p27和Cyclin D1的各自表达无明显差异;p27在正常卵巢组织和卵巢良性上皮肿瘤中高表达,在卵巢癌中表达降低(P<0.05),且随着肿瘤分级、分期增高(恶性程度增高),阳性表达率逐渐下降;而Cyclin D1的表达则相反;两者在肿瘤中的表达呈负相关.结论 p27kip表达下降、Cyclin D1过表达可能在卵巢癌的发生中起重要作用,检测p27kip1、Cyclin D1在卵巢癌中的表达可预测肿瘤生物学行为特征,可以作为判断预后的指标.     

幽门螺杆菌感染与胃癌中p21和p27表达的关系

探讨胃癌中p21~(WAF1)和p27~(KIP1)的表达及其临床意义,并分析幽门螺杆菌(Hp)感染和p21~(WAF1)、p27~(KIP1)表达的关系。采用免疫组化法检测了89例胃癌组织中p21~(WAF1)和p27~(KIP1)表达水平,并采用快速尿素酶试验和组织病理学检测两种方法检查这些胃癌病例的Hp感染情况。实验结果显示,胃癌组织p21~(WAF1)的表达水平在不同病例组织中有所差异。结合临床病理学指数分析显示,降低的p21~(WAF1)表达与较深的肿瘤侵袭密切相关(P<0.05)。免疫组织化学结果显示,p27~(KIP1)无论在细胞浆中还是细胞核内都有表达,p27~(KIP1)核表达水平与胃癌的组织病理学分型密切相关(P<0.05)。此外,还发现Hp阳性病例的p27~(KIP1)阳性表达率明显低于Hp阴性者(P<0.05),而p21~(WAF1)表达阳性率在Hp阳性和阴性胃癌病例中无显著差异(P>0.05)。结果提示,胃癌的进展与细胞周期调节蛋白p21~(WAF1)和p27~(KIP1)的表达下调有关,Hp感染的致癌过程中可能有p27~(KIP1)参与。     

应用组织芯片技术研究Survivin基因蛋白在前列腺肿瘤组织中的表达及其意义

目的:应用组织芯片技术分析Survivin基因蛋白在人类前列腺癌组织、前列腺正常组织及前列腺良性痛变组织中的表达情况.方法:采用兔抗人survivin单克隆抗体的免疫组织化学ABC法,研究Survivin在不同前列腺组织的表达,并分析Survivin在不同前列腺组织中的表达差异.结果:免疫组化结果显示,前列腺癌组织与前列腺良性病变组织及正常前列腺组织中Survivin的表达相比呈显著性差异(P<0.05).结论:Survivin在前列腺癌组织中呈高表达,提示其可能对前列腺癌的发生或发展有重要作用.     

p21和p27基因在眼睑皮脂腺腺癌中的表达及意义

目的探讨p21和p27基因在眼睑皮脂腺腺癌中的表达差异。方法收集武汉大学人民医院和武汉大学中南医院病理科2000-2008年手术切除及活检的眼睑皮脂腺腺癌(eyelid sebaceous gland grandular cancer,ESGGC)标本共20例,另取癌周围组织5例作对照。采用免疫组织化学方法观察各组组织内p21和p27基因的表达。并利用HPIAS-2000图像分析系统测定p21和p27基因在以上各组中表达的平均光密度和平均阳性面积率。对各组组织中p21和p27基因表达的阳性面积率作双变量相关分析。结果 1.p21和p27基因在眼睑皮脂腺腺癌组织中呈低表达;p21和p27基因在癌旁组织中呈高表达。经单因素方差分析,组间有显著性差异(P0.05?。经q检验,眼睑皮脂腺腺癌与癌旁组织之间,p21和p27基因表达的平均光密度及阳性面积率有显著性差异(P0.05?。2.p21和p27基因之间的表达呈显著正相关。结论1.p21和p27基因在眼睑皮脂腺腺癌组织中异常表达,对眼睑皮脂腺腺癌的发生和发展起了重要作用;2.p21和p27基因之间的表达呈显著正相关,它们在皮脂腺腺癌的发生和发展过程中可能起协同作用。     

Differential regulation of cyclin D1 and D3 expression in the control of astrocyte proliferation induced by endothelin-1

We have previously shown that the mitogenic effect of endothelin-1 (ET-1) in primary astrocytes is dependent on activation of both extracellular signal-regulated kinase (ERK)- and cytoskeleton (CSK)-dependent pathways. In this study, we evaluated the contribution of each of these pathways to the expression and activation of proteins mediating cell cycle progression. Our results suggest that ET-1-induced expression of cyclins D1 and D3 is dependent on the ERK- and CSK-dependent pathways, respectively; moreover, a decrease in the levels of the cyclin-dependent kinase inhibitor (CKI) p27 was observed as a consequence of ERK activation. Expression of both cyclins D1 and D3 together with a decrease in the p27 levels are essential for retinoblastoma protein (pRB) phosphorylation and cyclin A expression. Furthermore, the molecular events responsible for cell-cell contact inhibition of astrocyte proliferation were found to be independent of the mitogenic pathways leading to D-type cyclin expression. Cell growth arrest in confluent astrocytes was found to be correlated with increased expression of CKI p21, resulting in inhibition of D-type cyclin-associated pRB phosphorylation and cyclin A expression. Taken together, these results indicate that cyclins D1 and D3, which constitute the key mediators of the proliferative response of primary astrocytes to ET-1, are regulated by distinct signaling pathways.     

WWOX蛋白在胰腺癌组织中的表达及其临床意义

目的探讨WWOX蛋白在胰腺癌组织中的表达及其临床意义。方法应用免疫组织化学孓P法检测人胰腺癌组织芯片150芯（包括70例胰腺癌组织和5例胰腺炎）中WWOX蛋白的表达,采用HPIAS-1000图文报告管理系统对WWOX蛋白的表达进行定量分析,并用SPSS11．5软件对各组免疫组织化学反应阳性颗粒的平均光密度、阳性面积率做单因素方差分析和SNK（q）检验。结果胰腺癌组织中WWOX蛋白呈低表达,胰腺炎组织中WWOX蛋白呈高表达。结论提示WWOX基因表达下调可能在胰腺癌的发生、发展中起了重要作用。     

BRCA1-IRIS regulates cyclin D1 expression in breast cancer cells

The regulator of cell cycle progression, cyclin D1, is up-regulated in breast cancer cells; its expression is, in part, dependent on ERalpha signaling. However, many ERalpha-negative tumors and tumor cell lines (e.g., SKBR3) also show over-expression of cyclin D1. This suggests that, in addition to ERalpha signaling, cyclin D1 expression is under the control of other signaling pathways; these pathways may even be over-expressed in the ERalpha-negative cells. We previously noticed that both ERalpha-positive and -negative cell lines over-express BRCA1-IRIS mRNA and protein. Furthermore, the level of over-expression of BRCA1-IRIS in ERalpha-negative cell lines even exceeded its over-expression level in ERalpha-positive cell lines. In this study, we show that: (1) BRCA1-IRIS forms complex with two of the nuclear receptor co-activators, namely, SRC1 and SRC3 (AIB1) in an ERalpha-independent manner. (2) BRCA1-IRIS alone, or in connection with co-activators, is recruited to the cyclin D1 promoter through its binding to c-Jun/AP1 complex; this binding activates the cyclin D1 expression. (3) Over-expression of BRCA1-IRIS in breast cells over-activates JNK/c-Jun; this leads to the induction of cyclin D1 expression and cellular proliferation. (4) BRCA1-IRIS activation of JNK/c-Jun/AP1 appears to account for this, because in cells that were depleted from BRCA1-IRIS, JNK remained inactive. However, depletion of SRC1 or SRC3 instead reduced c-Jun expression. Our data suggest that this novel signaling pathway links BRCA1-IRIS to cellular proliferation through c-Jun/AP1 nuclear pathway; finally, this culminates in the increased expression of the cyclin D1 gene.     

Increased expression of MDM2, cyclin D1, and p27Kip1 in carcinogen-induced rat mammary tumors

It is thought that environmental pollutants, such as polycyclic aromatic hydrocarbons (PAH), contribute to human breast tumorigenesis, yet their roles remain incompletely elucidated. The prototypical PAH 7,12-dimethylbenz(alpha)anthracene (DMBA) specifically and effectively induces mammary tumor formation in rodent models. In an attempt to explore the molecular mechanisms by which PAH initiates and promotes mammary tumorigenesis, we examined the expression of several cell cycle regulators in rat mammary tumors induced by DMBA. Expression of cyclin D1, murine double minute-2 (MDM2), and Akt was up-regulated in tumors in comparison to normal mammary glands, as indicated by RT-PCR, Western blot analysis, and immunohistochemical staining. Expression of p27Kip1 protein was also elevated in the tumors with increased cytoplasmic localization. However, RB protein remained hyperphosphorylated. To directly test the effects of DMBA, the MCF-7 human breast cancer cells were treated. DMBA induced MDM2 expression in a dose- and time-dependent fashion in the MCF-7 cells, and this activation appeared to be p53 dependent. These data suggest that activation of cyclin D1, MDM2, and AKT as well as increased expression and cytoplasmic localization of p27Kip1 may play a role in this model of environmental pollutant-induced mammary tumorigenesis.     

P 7IkZP i、EF Z- 1在胃癌中的表达及其相关性研究

目的：研究p27^kip1蛋白、转录因子E2F-1在胃癌中的表达及其与胃癌病理参数乏问的关系,并探讨二者相关性。方法：应用免疫组化S-P法,对90例胃癌患者的癌组织、34例癌旁正常胃黏膜进行检测,分析p27^kip1、E2F-1蛋白表达及其与胃癌病理参数之间的关系。结果：p27^kip1在正常胃组织、胃癌组织中的阳性率分别为55．9％（19／34）、31．1％（28／90）,二者差异有统计学意义（p〈0．05）。其阳性表达率与胃癌组织的浸润深度（p〈0．05）、组织学分型（p〈0．01）、分化程度（p〈0．01）、淋巴结转移（p〈0．05）均相关;E2F-1在正常胃组织、胃癌组织中的阳性率分别为17．6％（6／34）、36．7％（33／90）,二者差异有统计学意义（p〈0．05）,其阳性表达率与胃癌的组织学分型相关。而与胃癌的浸润深度（p〉0．05）、分化程度（p〉0．05）、淋巴结转移无关（p〉0．05）。结论：p27^kip1在胃癌组织中的表达明显低于在正常组织,相反,E2F-1蛋白在胃癌组织中的表达却明显增强;p27^kip1蛋白表达与胃癌的浸润深度、组织学分型、分化程度、淋巴结转移等病理参数相关;E2F-1表达与胃癌的组织学分型相关;p27^kip1、E2F-1在胃癌中的表迭呈负相关。     

Influence of cyclin D1 splicing variants expression on breast cancer chemoresistance via CDK4/CyclinD1-pRB-E2F1 pathway

Cyclin D1 (CCND1), a mediator of cell cycle control, has a G870A polymorphism which results in the formation of two splicing variants: full-length CCND1 (CCND1a) and C-terminally truncated CCND1 species (CCND1b). However, the role of CCND1a and CCND1b variants in cancer chemoresistance remains unknown. Therefore, this study aimed to explore the molecular mechanism of alternative splicing of CCND1 in breast cancer (BC) chemoresistance. To address the contribution of G870A polymorphism to the production of CCND1 variants in BC chemoresistance, we sequenced the G870A polymorphism and analysed the expressions of CCND1a and CCND1b in MCF-7 and MCF-7/ADM cells. In comparison with MCF-7 cells, MCF-7/ADM cells with the A allele could enhance alternative splicing with the increase of SC-35, upregulate the ratio of CCND1b/a at both mRNA and protein levels, and activate the CDK4/CyclinD1-pRB-E2F1 pathway. Furthermore, CCND1b expression and the downstream signalling pathway were analysed through Western blotting and cell cycle in MCF-7/ADM cells with knockdown of CCND1b. Knockdown of CCND1b downregulated the ratio of CCND1b/a, demoted cell proliferation, decelerated cell cycle progression, inhibited the CDK4/CyclinD1-pRB-E2F1 pathway and thereby decreased the chemoresistance of MCF-7/ADM cells. Finally, CCND1 G870A polymorphism, the alternative splicing of CCDN1 was detected through Sequenom Mass ARRAY platform, Sanger sequencing, semi-quantitative RT-PCR, Western blotting and immunohistochemistry in clinical BC specimens. The increase of the ratio of CCND1b/a caused by G870A polymorphism was involved in BC chemoresistance. Thus, these findings revealed that CCND1b/a ratio caused by the polymorphism is involved in BC chemoresistance via CDK4/CyclinD1-pRB-E2F1 pathway.     

The Down syndrome-related protein kinase DYRK1A phosphorylates p27Kip1 and Cyclin D1 and induces cell cycle exit and neuronal differentiation

A fundamental question in neurobiology is how the balance between proliferation and differentiation of neuronal precursors is maintained to ensure that the proper number of brain neurons is generated. Substantial evidence implicates DYRK1A (dual specificity tyrosine-phosphorylation-regulated kinase 1A) as a candidate gene responsible for altered neuronal development and brain abnormalities in Down syndrome. Recent findings support the hypothesis that DYRK1A is involved in cell cycle control. Nonetheless, how DYRK1A contributes to neuronal cell cycle regulation and thereby affects neurogenesis remains poorly understood. In the present study we have investigated the mechanisms by which DYRK1A affects cell cycle regulation and neuronal differentiation in a human cell model, mouse neurons, and mouse brain. Dependent on its kinase activity and correlated with the dosage of overexpression, DYRK1A blocked proliferation of SH-SY5Y neuroblastoma cells within 24 h and arrested the cells in G₁ phase. Sustained overexpression of DYRK1A induced G₀ cell cycle exit and neuronal differentiation. Furthermore, we provide evidence that DYRK1A modulated protein stability of cell cycle-regulatory proteins. DYRK1A reduced cellular Cyclin D1 levels by phosphorylation on Thr286, which is known to induce proteasomal degradation. In addition, DYRK1A phosphorylated p27^Kip1 on Ser10, resulting in protein stabilization. Inhibition of DYRK1A kinase activity reduced p27^Kip1 Ser10 phosphorylation in cultured hippocampal neurons and in embryonic mouse brain. In aggregate, these results suggest a novel mechanism by which overexpression of DYRK1A may promote premature neuronal differentiation and contribute to altered brain development in Down syndrome.