细胞周期蛋白在喉癌组织中的表达及意义

目的探讨喉癌组织中细胞周期蛋白（Cyclin D1、Cyclin A、Cyclin B1、Cyclin E）的表达规律及其与喉癌临床病理参数的关系。方法采用免疫组化技术,检测Cyclin D1、Cyclin A、Cyclin B1、Cyclin E在67例喉癌组织和32例癌旁正常组织中的表达情况。结果 Cyclin D1、Cyclin A、Cyclin B1、Cyclin E在癌旁正常组织中呈低表达。随着喉癌分化程度的降低,Cyclin D1、Cyclin A、Cyclin B1、Cyclin E表达阳性率明显升高,高分化者、中分化者与低分化者比较有显著性差异（P〈0.05）。结论通过免疫组织化学法分析细胞周期的数据,有利于肿瘤分级,且细胞周期数据与预后相关。     

急性白血病患者细胞周期蛋白B1基因表达与耐药的关系

目的:研究急性白血病(AL)患者细胞周期蛋白B1基因的表达与耐药的关系。方法:应用半定量逆转录多聚酶链反应(RTPCR)检测了13例耐药急性白血病患者和14例非耐药急性白血病患者外周血细胞周期蛋白B1基因的表达水平,并应用荧光定量RTPCR对所有患者外周血mdr1基因的表达进行了检测。结果:耐药组细胞周期蛋白B1基因表达水平显著高于非耐药组(P<0.05,t=2.549),细胞周期蛋白B1与mdr1mRNA的表达呈正相关(r=0.404,P<0.05,t=2.208)。结论:细胞周期蛋白B1基因可以作为判断AL患者耐药的指标,对判断AL的预后具有一定的价值,细胞周期蛋白B1的表达与由mdr1介导的耐药相关。     

细胞周期蛋白D1表达与宫颈鳞癌关系的研究

目的探讨细胞周期蛋白D1(cyclin D1)表达与宫颈鳞癌关系的规律,为寻找一种早期诊断的手段奠定基础.方法采用班点杂交方法检测96例经福尔马林固定,石蜡包埋的宫颈鳞状上皮癌(宫颈鳞癌)38例,宫颈鳞状上皮不典型增生32例、慢性宫颈炎12例和正常宫颈组织标本14例中cyclin D1的表达情况.结果cyclin D1的表达在宫颈鳞癌与不典型增生相差不显著(P＞0.05).显著高于慢性宫颈炎和正常宫颈组织(P＜0.05).结论揭示cyclin D1的高表达与宫颈鳞癌的发生有关.     

胃癌组织中端粒酶活性与细胞周期蛋白D_1表达关系的研究

目的　探索胃癌组织中端粒酶活性与细胞周期蛋白D1表达之间可能的关系。方法　利用TelomerasePCR ELISAKit测量端粒酶活性 ;利用RT PCRELISA检测细胞周期蛋白D1mRNA表达水平。共检测了 84例胃癌及其相关性的组织标本。结果　周期蛋白D1异常高表达在胃良性疾病、胃癌前期病变和胃癌组织中均可检出 ,而端粒酶活性仅在胃癌前期病变和胃癌组织中检出。胃癌前期病变和胃癌组织端粒酶活性与周期蛋白D1表达程度明显正相关 (r =0 .713,P <0 .0 5 )。结论　至少在胃癌组织中 ,细胞周期蛋白D1异常高表达可能是端粒酶激活的前提之一。     

应用细胞周期阻断法对白血病骨髓及外周血细胞的微核分析

本文应用细胞周期阻断法,对51例白血病患者骨髓,30例白血病患者外周血细胞,19例非白血病患者骨髓和30例正常献血员外用血细胞进行了微核分析。结果显示,外周血细胞微核率,白血病患者明显高于正常献血员;骨髓细胞微核率,白血病患者明显高于非白血病患者;同一病人骨髓细胞和外周血细胞微核率无差异性。     

非小细胞肺癌中细胞周期蛋白D1与E的作用及相互关系

目的 :研究细胞周期蛋白D1与E在非小细胞肺癌 (NSCLC)发生、发展中的作用及相互关系。方法 :免疫组化方法检测 87例NSCLC肿瘤组织中细胞周期蛋白D1、E的表达及PCNA估计增殖指数 (PI) ,并将上述结果与临床病理及预后资料进行对比分析。结果 :87例NSCLC中细胞周期蛋白D1、E阳性率分别为 4 4 .8% (39/87)、4 8.3% (42 /87) ,细胞周期蛋白D1阳性组的PI值显著高于阴性组 (P〈0 .0 5 ) ,细胞周期蛋白E阳性组PI值与阴性组无显著差异 (P〉0 .0 5 ) ;细胞周期蛋白D1阳性组肿瘤直径、淋巴结转移率和生存率均与阴性组有显著差异(P〈0 .0 1、〈0 .0 5、〈0 .0 1) ,细胞周期蛋白E阳性组淋巴结转移率、临床分期和生存率均与阴性组有显著差异 (P〈0 .0 5、〈0 .0 5、〈0 .0 1) ;细胞周期蛋白D1阳性组中细胞周期蛋白E的阳性率显著高于其阴性组 (P〈0 .0 5 ) ;细胞周期蛋白D1与细胞周期蛋白E双阳性组的PI值、肿瘤直径、淋巴结转移率显著高于非双阳性组 (P值均〈0 .0 5 ) ,生存率显著低于非双阳性组 (P〈0 .0 1)。结论 :细胞周期蛋白D1、细胞周期蛋白E均参与NSCLC的发生、发展 ,并影响其预后 ,但两者在其中所起作用不同 :细胞周期蛋白D1是调节NSCLC增殖的主要因素 ,细胞周期蛋白E主要与NSCLC进展有关 ;细胞周期蛋白D1可促     

细胞周期蛋白D1表达与乳腺癌新辅助化疗关系的研究

目的 观察乳腺癌新辅助化疗(NAC)前后cyclin D1表达情况,探讨其与化疗效果之间的关系.方法 选择84例乳腺癌患者经空心针穿刺获取组织学样本并加以验证,经3～4个周期ET方案[吡柔比星(THP)+多西紫杉醇(TXT)]NAC,采用免疫组织化学Envision二步法检测化疗前、后cyclinD1的表达变化情况.结果 84例患者中完全缓解(CR)4例(4.76％)(其中病理CR 2例),部分缓解(PR) 54例(64.29％),稳定(SD) 26例(30.95％),无疾病进展(PD)患者.NAC后乳腺癌组织中cyclin D1表达降低[65.48％(55/84)],与化疗前[39.29％(33/84)]相比,差异有统计意义(x 2=11.55,P=0.001).NAC后cyclin D1阳性表达转为阴性表达者临床缓解率[86.36％(19/22)]高于未转为阴性表达者[45.45％(15/33)],差异有统计学意义(x2=9.359,P=0.002).结论 NAC降低乳腺癌组织中的cyclinD1表达,且NAC后cyclinD1表达转阴者化疗效果提高,可以评估NAC的有效性.     

细胞周期蛋白与急性白血病预后的关系研究

背景与目的:细胞的增殖和分化是通过细胞周期转运来实现的,当肿瘤细胞异常增殖时,作为细胞运转正调因子的细胞周期蛋白(cyclins)就有可能表达异常。目前已有较多资料证实cyclins在实体肿瘤中呈高表达。本研究探讨cyclins与急性白血病预后的关系。方法:实验组分为急性白血病完全缓解组(12例)、急性白血病初发未治组(16例)、难治性白血病组(40例);正常对照组为良性血液病患者(15例)。用RT-PCR法检测cyclinA、cyclinD、cyclinE。结果:正常对照组15例的cyclins表达均为阴性,在正常对照组和实验组中,cyclinA的表达没有显著性差异(P>0.05),而cyclinD和cyclinE在实验组的表达均明显高于正常对照组(P<0.01)。在急性白血病完全缓解组,cyclinD和cyclinE的表达无显著性差异(P>0.05);而在难治性白血病中,cyclinD的表达显著高于cyclinE,且cyclinD在难治性白血病组的表达明显高于初发性的病例(P<0.05)。将所有cyclins表达阳性的病例分为表达单一cyclin和表达2个以上cyclins两组,并分别计算两者的阳性率,发现两者的阳性率在急性白血病完全缓解组和难治性白血病组间无显著性差异(P>0.05)。结论:CyclinD可以作为急性白血病的预后指标,不能用cyclins基因类型表达的多寡来判断预后。     

细胞周期蛋白D1基因对卵巢癌细胞生物学行为的影响#br#

【摘要】目的基于Oncomine数据库分析细胞周期蛋白D1（Cyclin D1）基因在卵巢癌中的表达水平及沉默其表达对癌细胞生物学行为的影响。方法利用Oncomine数据库中关于卵巢癌组织中Cyclin D1基因的相关信息,分析Cyclin D1基因在不同肿瘤中的表达情况以及在卵巢癌中的表达情况。使用在线数据库Kaplan Meier Plotter分析Cyclin D1表达水平与卵巢癌预后的关系。使用qRT PCR检测各卵巢癌细胞株（SKOV3、CAOV3、A2780、ES2）及正常卵巢上皮细胞中Cyclin D1的表达。取对数生长期卵巢癌细胞SKOV3,将细胞分为空白对照组、阴性对照组（NC siRNA组）和Cyclin D1基因沉默组（Cyclin D1 siRNA组）;使用Western blotting检测各组Cyclin D1蛋白表达;分别使用MTT法、细胞划痕实验和Transwell实验检测各组SKOV3细胞的增殖、迁移和侵袭能力。结果通过挖掘分析Oncomine数据库涉及的Cyclin D1基因发现,与正常卵巢组织比较,在所有差异表达基因中,Cyclin D1基因的中位数值排名为2020,P<0001。与Cyclin D1低表达的卵巢癌患者比较,Cyclin D1高表达患者的总生存期和无瘤进展生存期均较短,差异有统计学意义（P<001）。与正常卵巢上皮细胞比较,Cyclin D1基因在各卵巢癌细胞（ES2、CAOV3、SKOV3、A2780）中的表达明显升高,差异有统计学意义（P<005）;与SKOV3细胞株比较,Cyclin D1基因在CAOV3、A2780、ES2细胞株中的表达明显降低,差异有统计学意义（P<005）。Cyclin D1 siRNA组细胞中Cyclin D1的表达量、增殖能力、迁移面积、穿过聚碳酸酯膜的癌细胞数量均明显低于空白对照组和NC siRNA组,差异有统计学意义（P<005）。结论Cyclin D1基因在卵巢癌组织中高表达,该基因可能参与卵巢癌的发生发展和转移,并与患者预后密切相关。     

细胞周期蛋白D1在非小细胞肺癌中表达的临床意义

目的 探讨非小细胞肺癌（ＮＳＣＬＣ）中细胞周期蛋白Ｄ１（ＣｙｃｌｉｎＤ１）的表达及其临床意义。方法 用鼠抗人ＣｙｃｌｉｎＤ１单克隆抗体（ＤＣＳ／６）对８９例石蜡包埋的原发性ＮＳＣＣＬ组织进行免疫组化染色，并与２０例肺炎性病变比较。结果 ＣｙｃｌｉｎＤ２在肺鳞癌、腺癌、大细胞癌中都有较高的表达，其阳性率分别为６０．５％、６１．０％、４０．０％；而在肺炎性病变中，在肺鳞癌、腺癌中，肿块直径≥３ｃｍ、有     

洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和周期的影响

目的： 探讨洋地黄毒苷对肺癌NCI-H446与A549细胞增殖和细胞周期的影响,并初步探讨其可能的作用机制。方法：体外培养的NCI-H446与A549细胞分别经不同浓度(10、20、40、80、160 nmol/L)的洋地黄毒苷处理24、48和72 h后,采用MTT法检测细胞增殖情况,应用流式细胞术检测洋地黄毒苷处理细胞48 h后各组细胞周期分布,Western blot检测细胞中Cyclin A和P21蛋白表达水平。结果：与未经洋地黄毒苷处理的对照组相比,不同浓度(10、20、40、80和160 nmol/L)的洋地黄毒苷均可抑制 NCI-H446与A549细胞的增殖, 均呈剂量和时间依赖性(P<0.05),作用48 h后,洋地黄毒苷对NCI-H446与A549细胞的IC50值分别为61.26 nmol/L和110.73 nmol/L。流式细胞仪分析结果显示,随药物作用浓度增加,G0/G1细胞比例降低,S期细胞比例显著增加,与对照组相比,差异具有统计学意义(P<0.05)。Western blot分析结果显示,洋地黄毒苷能剂量依赖性的下调Cyclin A1蛋白表达和上调P21 蛋白的表达(P<0.05)。结论：洋地黄毒苷可抑制体外培养的肺癌NCI-H446与A549细胞增殖,诱导细胞发生S期阻滞,其机制可能与细胞周期相关调控蛋白的表达有关。     

细胞周期检测点的流式细胞术分析

背景与目的:真核细胞的细胞周期运行遵循着严格的时相次序,这种精密的延缓和强制的次序,由细胞监控机制——检测点来完成,检测点功能的丧失会导致肿瘤的发生。传统的细胞周期检测点分析多数基于群体细胞DNA直方图的流式细胞术。本研究以晚G1期检测点为模式,建立一种新的细胞周期检测点分析方法——Cyclins/DNA双参数流式细胞术,并评价应用该方法进行细胞周期检测点分析的可行性和优越性。方法:将紫外线照射诱导后的人类急性淋巴细胞型白血病细胞株(MOLT-4)于不同时间点收获固定,分两组进行流式细胞仪检测:一组用DNA直方图法,Modifit软件分析计算G0/G1期细胞总数;另一组应用CyclinE/DNA双参数流式细胞术对G1晚期细胞CyclinE的荧光强度、阈值及G0/G1各亚期细胞数量进行定量分析,并比较两组实验结果。结果:用DNA直方图法观察到紫外线照射后0～4hG0/G1期细胞总数基本不变(5300～5500),6h后开始上升(6241,12.6%)。而用CyclinE/DNA双参数法观察到:(1)G1晚期细胞的CyclinE荧光强度于照射后短时间内便开始变化,1h上升为341.2(对照295.1,15.6%),6h上升为577.6(95.7%),此时CyclinE阈值上升到5.4(0h为2.0);(2)G0/G1各亚期细胞数目变化趋势不明显:G1晚期细胞数6h时略为减少(此时凋亡率为5.61%),G1早期细胞数缓慢上升。结论:CyclinE/DNA双参数流式细胞术将CyclinE的荧光强度及表达阈值定量化,对于细胞晚G1期检测点的检测比传统的DNA直方图法更为敏感和精确。     

EHMT1 knockdown induces apoptosis and cell cycle arrest in lung cancer cells by increasing CDKN1A expression

Dozens of histone methyltransferases have been identified and biochemically characterized, but the pathological roles of their dysfunction in human diseases such as cancer remain largely unclear. Here, we demonstrate the involvement of EHMT1, a histone lysine methyltransferase, in lung cancer. Immunohistochemical analysis indicated that the expression levels of EHMT1 are significantly elevated in human lung carcinomas compared with non‐neoplastic lung tissues. Through gene ontology analysis of RNA‐seq results, we showed that EHMT1 is clearly associated with apoptosis and the cell cycle process. Moreover, FACS analysis and cell growth assays showed that knockdown of EHMT1 induced apoptosis and G1 cell cycle arrest via upregulation of CDKN1A in A549 and H1299 cell lines. Finally, in 3D spheroid culture, compared to control cells, EHMT1 knockdown cells exhibited reduced aggregation of 3D spheroids and clear upregulation of CDKN1A and downregulation of E‐cadherin. Therefore, the results of the present study suggest that EHMT1 plays a critical role in the regulation of cancer cell apoptosis and the cell cycle by modulating CDKN1A expression. Further functional analyses of EHMT1 in the context of human tumorigenesis may aid in the development of novel therapeutic strategies for cancer.     

Monensin-mediated growth inhibition in acute myelogenous leukemia cells via cell cycle arrest and apoptosis

Monensin, an Na(+) ionophore, regulates many cellular functions including apoptosis. However, there has been no report about the antitumoral effect of monensin on acute myelogenous leukemia (AML). Here, we investigated the antiproliferative effect of monensin on AML cells in vitro and in vivo. Monensin efficiently inhibited the proliferation of all of 10 AML cell lines, with IC(50) of about 0.5 microM. DNA flow cytometric analysis indicated that monensin induced a G(1) and/or a G(2)-M phase arrest in these cell lines. To address the mechanism of the antiproliferative effect of monensin, we examined the effect of monensin on cell cycle-related proteins in HL-60 cells. The levels of CDK6, cyclin D1 and cyclin A were decreased. In addition, monensin not only increased the p27 level but also enhanced its binding with CDK2. Furthermore, the activities of CDK2- and CDK6-associated kinases reduced by monensin were associated with hypophosphorylation of Rb protein. Monensin also induced apoptosis in AML cells including HL-60 cells. The apoptotic process of HL-60 cells was associated with changes in Bax, caspase-3, caspase-8 and mitochondria transmembrane potential (Deltapsi(m)). In particular, monensin (i.p. at a dose of 8 mg/kg thrice weekly) significantly reduced the tumor size of BALB/c mice that were inoculated s.c. with its derived cell line, WEHI-3BD cells (69% growth inhibition relative to control group; p < 0.05). Tumors from monensin-treated mice exhibited increased apoptosis, and these tumor were immunohistochemically more stained with Bax, Fas and p53 antibodies than control tumors. In conclusion, this is the first report that monensin potently inhibits the proliferation of AML cells.     

2-Methoxyestradiol and paclitaxel have similar effects on the cell cycle and induction of apoptosis in prostate cancer cells

2-Methoxyestradiol (2-ME) is an endogenous metabolite of estradiol with promise for cancer chemotherapy, including advanced prostate cancer. We have focused on events related to cell cycle arrest (G1 and G2/M) and induction of apoptosis in human prostate cancer cells. Treatment with 2-ME increased cyclin B1 protein and its associated kinase activity followed by later inhibition of cyclin A-dependent kinase activity and induction of apoptosis. Similar results were obtained with paclitaxel (taxol), a clinically relevant agent used to treat advanced prostate cancer. Cyclin-dependent kinase inhibitors prevented 2-ME and paclitaxel-mediated increase in cyclin B1-dependent kinase activity and blocked induction of apoptosis. Reduction of X-linked inhibitor of apoptosis (XIAP) protein by 2-ME and paclitaxel correlated with increased apoptosis. Lower doses of 2-ME and paclitaxel resulted in G1 (but not G2/M) cell cycle arrest in the p53 wild type LNCaP cell line, but with minimal induction of apoptosis. We suggest that 2-ME and paclitaxel-mediated induction of apoptosis in prostate cancer cells requires activation of cyclin B1-dependent kinase that arrests cells in G2/M and subsequently leads to the induction of apoptotic cell death.     

PBRM1 suppresses bladder cancer by cyclin B1 induced cell cycle arrest

Growing evidence indicates that dys-regulation of PBRM1 contributes to tumorigenesis. However, little is known about the biological function of PBRM1 in the development or progression of bladder cancer. In this study, we aimed to elucidate the pathophysiological role of PBRM1 in bladder cancer. We assessed the expression of PBRM1 in 64 bladder cancer tissue samples with matching normal tissues. We explored the biological functions of PBRM1 both in vitro and in vivo. Mutational status of PBRM1 was analyzed. Effect of PBRM1 on cell cycle was evaluated. qRT-PCR and Western blot were carried out to evaluate the expression of cyclins affected by PBRM1. Our results showed that PBRM1 expression was significantly reduced in bladder cancer cells and tissues compared to their normal counterparts. The reduced expression of PBRM1 was associated with advanced tumor stage, low differentiation grade and worse patient outcome. Further functional analysis demonstrated that PBRM1 suppressed bladder cancer cell proliferation, migration, colony formation in vitro and tumorigenicity in vivo. Genetic alteration analysis showed no amino-acid sequence altering mutations. We found that PBRM1 could block the G2/M transition by repressing cyclin B1. Our data indicated that PBRM1 functions as a tumor suppressor in bladder cancer by repressing cyclin B1 expression.     

蟾蜍灵对胃癌组织MGC-803细胞周期作用机制的研究

目的探讨蟾蜍灵对胃癌MGC803细胞周期及周期蛋白的影响。方法采用台盼蓝拒染法测定细胞增殖活力;瑞士姬姆萨染色观察细胞形态学的变化;流式细胞仪检测细胞周期;免疫组织化学方法检测细胞周期相关蛋白p16、p21、pRb的表达。结果1)蟾蜍灵抑制胃癌MGC803细胞的增殖,24、48、72h的IC50分别为0.086、0.048和0.036μmol/L。2)蟾蜍灵在0.01～0.1μmol/L时作用24～72h,形态学上可见细胞体积增大,出现双核、多核细胞。3)流式细胞仪显示蟾蜍灵浓度为0.01～0.1μmol/L时可明显诱导细胞周期G2/M期阻滞。4)蟾蜍灵作用于胃癌MGC803细胞,观察到p16、p21、pRb蛋白的表达明显上调。结论蟾蜍灵引起胃癌MGC803细胞的周期阻滞可能与p16、p21、pRb蛋白的上调有关。     

Exogenous morphine inhibits human gastric cancer MGC- 803 cell growth by cell cycle arrest and apoptosis induction

Morphine is not only an analgesic treating pain for patients with cancer but also a potential anticancer drug inhibiting tumor growth and proliferation. To gain better insight into the involvement of morphine in the biological characteristics of gastric cancer, we investigated effects on progression of gastric carcinoma cells and the expression of some apoptosis-related genes including caspase-9, caspase-3, survivin and NF-κB using the MGC-803 human gastric cancer cell line. The viability of cells was assessed by MTT assay, proliferation by colony formation assay, cell cycle progression and apoptosis by flow cytometry and ultrastructural alteration by transmission electron microscopy. The influences of morphine on caspase-9, caspase-3, survivin and NF-κB were evaluated by semi-quantitative RT-PCR and Western blot. Our data showed that morphine could significantly inhibit cell growth and proliferation and cause cell cycle arrest in the G2/M phase. MGC-803 cells which were incubated with morphine also had a higher apoptotic rate than control cells. Morphine also led to morphological changes of gastric cancer cells. The mechanism of morphine inhibiting gastric cancer progression in vitro might be associated with activation of caspase-9 and caspase-3 and inhibition of survivin and NF-κB.