膀胱癌组织中CXCR4 和CXCR7 的表达及临床意义

目的:探讨膀胱癌组织中趋化因子受体4(CXCR4)和趋化因子受体7(CXCR7)的表达及临床意义。方法:收集2012年1月至2014年1月我院收集的膀胱癌组织标本96例,肿瘤旁正常组织标本42例,采用免疫组化方法检测组织标本中CXCR4和CXCR7的表达情况。结果:96例癌组织中检出CXCR4阳性59例,阳性率为61.46%,检出CXCR7阳性表达71例,阳性率为73.96%;42例癌旁组织中检出CXCR4阳性11例,阳性率26.19%,检出CXCR7阳性8例,阳性率为19.05%,癌组织与癌旁组织中CXCR4和CXCR7的表达具有统计学差异(均P0.05);相关性分析显示在膀胱癌组织中,CXCR4和CXCR7的表达呈正相关性(r=0.497,P=0.001);CXCR4和CXCR7在浸润性高(T2-T3)的膀胱癌和分化程度低(G2-G3)的膀胱癌表达强度较高,且差异具有统计学意义(均P0.05)。结论:CXCR4和CXCR7协同参与了膀胱癌的发生发展,并且与肿瘤的分化程度和浸润程度密切相关,有望成为诊断和治疗的重要靶点,在临床应用上具有重要意义。     

eIF-4E 和MMP9 在子宫内膜癌中的表达及临床意义

目的:探讨真核翻译起始因子4E(e IF-4E)和基质金属蛋白酶9(MMP9)在子宫内膜癌组织中的表达及其临床意义。方法:应用免疫组化法检测e IF-4E和MMP9在16例正常子宫内膜组织、10例增生子宫内膜组织及42例子宫内膜癌组织中的表达,分析其与子宫内膜癌临床病理之间的关系,并研究二者的相关性。结果:e IF-4E与MMP9在三组组织中的表达阳性率逐渐增加,三组间的差异有统计学意义(P0.05),两两比较,正常内膜和内膜癌之间的差异有统计学意义(P0.0167)。e IF-4E和MMP9的表达与子宫内膜癌的FIGO分期、组织学分级及淋巴结转移具有显著相关性(P0.05);子宫内膜癌组织中e IF-4E与MMP9的阳性表达呈正相关(r=0.717,P0.05)。结论:e IF-4E和MMP9在子宫内膜癌组织中表达显著上调,随FIGO分期的增加、组织学分级的增高及有淋巴结转移而增高,可能与子宫内膜癌的发生发展相关,二者的联合检测对于评估子宫内膜癌患者的病情及预后有重要的参考意义。     

应用套叠PCR技术构建番木瓜eIF4E和eIFiso4E基因的hpRNA植物表达载体

植物真核翻译起始因子4E(eIF4E)在蛋白质合成的起始中发挥重要作用,参与植物-病毒互作,影响病毒的侵染过程.为了研究eIF4E在植物病毒侵染中的功能,建立了一种快速的套叠PCR新方法,成功构建了番木瓜eIF4E和eIFiso4E基因的hpRNA结构,并将其连接到改造的植物表达载体pBI121上,为利用RNA干扰技术研究番木瓜eIF4E和eIFiso4E基因在病毒侵染中的作用奠定了基础.     

胃癌组织中Bmi-1基因表达的研究

目的：多梳基因家族的Bmi．1被认为是一种癌基因,在多种肿瘤组织中均有表达。本研究主要是检测Bmi-1基因在胃癌组织中的表达及探讨其临床意义。方法：应用RT—PCR和Westernblotting方法检测45例胃癌中Bmi-1基因的表达情况,并结合患者的临床病理资料分析与其相关性。结果：Bmi-1基因阳性表达率在胃癌中为88．9％（40／45）,癌旁组织为17．7％（8／45）,差异有显著性（P〈0．05）;Bmi-1基因阳性表达率与患者年龄、性别、肿瘤大小及有无淋巴结转移无关,与肿瘤分化程度及TNM分期有关（P〈0．05）。结论：Bmi-1基因在胃癌中高表达,与胃癌的疾病进展密切相关,检测Bmi-1的表达可作为胃癌生物学行为的一项评估指标。     

胃癌组织中Bmi-1基因表达的研究

目的:多梳基因家族的Bmi-1被认为是一种癌基因,在多种肿瘤组织中均有表达。本研究主要是检测Bmi-1基因在胃癌组织中的表达及探讨其临床意义。方法:应用RT-PCR和Western blotting方法检测45例胃癌中Bmi-1基因的表达情况,并结合患者的临床病理资料分析与其相关性。结果:Bmi-1基因阳性表达率在胃癌中为88.9%(40/45),癌旁组织为17.7%(8/45),差异有显著性(P<0.05);Bmi-1基因阳性表达率与患者年龄、性别、肿瘤大小及有无淋巴结转移无关,与肿瘤分化程度及TNM分期有关(P<0.05)结论:Bmi-1基因在胃癌中高表达,与胃癌的疾病进展密切相关,检测Bmi-1的表达可作为胃癌生物学行为的一项评估指标。     

番木瓜eIF4E和eIFiso4E基因嵌合hpRNA载体一步快速构建及沉默效果的研究

番木瓜环斑病毒(Papaya ringspot virus,PRSV)严重威胁番木瓜种植业的发展,且目前没有十分有效的防治办法。病毒侵染植物依赖寄主因子的协助,真核翻译起始因子4E(eukaryotic initiation factor 4E,eIF4E)是多种RNA病毒侵染植物的必需因子。以番木瓜eIF4E家族基因为研究对象,构建同时干扰其eIF4E和eIFiso4E基因的发卡RNA(hairpin RNA,hpRNA)载体,并将其导入到番木瓜叶肉原生质中。通过荧光实时定量检测发现,番木瓜中eIF4E和eIFiso4E基因的表达量分别下降了49.8%和67.6%,这为进一步研究番木瓜eIF4E家族基因对PRSV侵染的影响以及利用RNA干扰技术靶向植物基因的病毒防治新策略提供理论和实践依据。     

B7-H4和FOXP3在乳腺癌中的表达及临床意义

目的:研究B7-H4和FOXP3在乳腺癌中的表达及其相互关系和临床意义.方法:应用S-P免疫组化法检测B7-H4蛋白和FOXP3蛋白在11例正常乳腺、25例乳腺良性病变、272例乳腺癌组织中的表达情况;运用原位杂交检查B7-H4 mRNA在10例正常乳腺、10例良性病变、20例乳腺癌组织中的表达情况.结果:B7-H4蛋白在乳腺癌中的阳性表达率(83.39%)高于乳腺良性病变组(64.00%)和乳腺正常组(54.55%),差异有统计学意义(p＜0.05);FOXP3在乳腺癌中的阳性表达率(82.67%)高于乳腺良性病变组(60.00%)和乳腺正常组(54.55%),差异有统计学意义(p＜0.05);B7-H4和FOXP3在乳腺癌中的阳性表达率与患者年龄、肿块大小和组织学分型无关(p＞0.05),而与组织学分级和淋巴结转移相关(p＜0.05);B7-H4和FOXP3在乳腺癌组织中的表达呈正相关(r=0.306,p＜0.001);B7-H4 mRNA在正常乳腺组、乳腺良性病变组、乳腺癌组中的阳性表达率差异没有统计学意义(p＞0.05);B7-H4 mRNA在乳腺癌中的阳性表达率与年龄、肿块大小、组织学分级、淋巴结转移无关(p＞0.05).结论:B7-H4和FOXP3在乳腺癌中表达上调,可能与乳腺癌的转移和预后相关.     

口蹄疫病毒非结构蛋白3AB 的原核表达及应用

旨在建立一种检测口蹄疫病毒非结构蛋白抗体的敏感、特异的ELISA方法。克隆、表达了口蹄疫病毒非结构蛋白3AB基因,原核表达的重组蛋白经亲和层析法纯化及Western blotting鉴定后作为包被抗原,建立检测口蹄疫病毒非结构蛋白抗体的3AB间接ELISA方法,通过与商品化试剂盒3ABC-ELISA的比对试验对其进行评价。结果显示,重组蛋白3AB以包涵体形式表达;能与口蹄疫病毒感染血清发生特异性反应,而不能与疫苗免疫动物血清发生反应;在检测田间样品时,与3ABC-ELISA具有同样的特异性和敏感性 (P＞     

E-cadherin和KLF 4表达对胃癌侵袭转移的作用

观察E-cadherin,Krüppel-like factor 4(KLF4)蛋白在胃癌和正常胃黏膜组织中的表达,分析其与胃癌浸润、转移的关系.应用免疫组织化学SP法检测84例手术切除的胃癌标本及对应正常胃黏膜组织中E-cadherin,KLF4蛋白的表达.各指标之间相关因素的差异性比较采用χ2检验,E-cadherin,KLF4相关性研究采用Spearman相关分析.结果显示,与正常胃组织相比,E-cadherin、KLF4蛋白在胃癌组织中均呈低表达或者缺失(分别42.9%vs.95.24%,8.3%vs 81%,P<0.05).E-cadherin、KLF4蛋白的阳性表达率与组织分级(P<0.05)、肿瘤浸润深度(P<0.05)、淋巴转移(P<0.05)明确相关.Spearman相关分析显示KLF4蛋白与E-cadherin蛋白的表达呈正相关(P<0.05).因此,E-cadherin,KLF4蛋白水平低表达可能与胃癌浸润和转移有关,而联合检测更能有效判断胃癌这一生物学行为.     

肠球菌溶血及非溶血菌株对小鼠巨噬细胞表达TNF-a的影响

【目的】探讨肠球菌溶血菌株及非溶血菌株对小鼠巨噬细胞RAW264.7表达TNF-а的影响。【方法】用多粘菌素B抑制排除内毒素污染对实验的影响。肠球菌溶血菌株、非溶血菌株各11株,以菌/细胞比 30∶1 感染RAW264.7细胞1 h,加入200 mg/mL氨苄青霉素继续培养24 h,分别于感染后3、6、9、24 h,用ELISA方法检测不同观测点细胞培养液中肿瘤坏死因子TNF-a的含量,并用逆转录-聚合酶链反应方法（RT-PCR）比较肠球菌溶血、非溶血菌株感染6 h后TNF-a mRNA表达的差异。【结果】未感染的RAW264.7细胞培养液中检测不到TNF-a。肠球菌溶血株感染组细胞培养上清液中各观测点的TNF-a的平均含量 (pg/mL) 均比非溶血株性组高。经t检验,P<0.01,差别有显著性。RT-PCR法检测其mRNA的表达也有相同结果：TNF-a mRNA在肠球菌溶血株感染细胞中的相对表达量比非溶血株感染的细胞高,经t检验,P<0.05,差别有统计学意义。【结论】肠球菌溶血株比非溶血株更能促进小鼠巨噬细胞RAW264.7产生TNF-а炎症因子。     

Differences in degradation lead to asynchronous expression of cyclin E1 and cyclin E2 in cancer cells

Cyclin E1 is expressed at the G₁/S phase transition of the cell cycle to drive the initiation of DNA replication and is degraded during S/G₂M. Deregulation of its periodic degradation is observed in cancer and is associated with increased proliferation and genomic instability. We identify that in cancer cells, unlike normal cells, the closely related protein cyclin E2 is expressed predominantly in S phase, concurrent with DNA replication. This occurs at least in part because the ubiquitin ligase component that is responsible for cyclin E1 downregulation in S phase, Fbw7, fails to effectively target cyclin E2 for proteosomal degradation. The distinct cell cycle expression of the two E-type cyclins in cancer cells has implications for their roles in genomic instability and proliferation and may explain their associations with different signatures of disease.     

GL331-induced disruption of cyclin B1/CDC 2 complex and inhibition of CDC 2 kinase activity

GL331, a new homologue of etoposide (VP-16), was developed to cope with the multiple drug resistance occurring in certain malignant tumours. We previously indicated that GL331, like VP-16 and other major cancer chemotherapeutic agents, induced apoptosis in a variety of human cancer cell lines including nasopharyngeal carcinoma (NPC) NPC-TW01 and NPC-TW04 cells. In this study, we further explored the effect of GL331 on the cell cycle progression of NPC cells. Flow cytometric analysis of DNA content was first used to demonstrate the ability of GL331 to induce cell growth arrest at S-G₂ phase in most NPC cells. Besides acting as a topoisomerase II inhibitor, GL331 inhibited cellular cyclin B1-associated CDC 2 kinase activity 6 h after treatment, accounting partly at least for its induction of the cell cycle arrest. As with cyclin A, D1, E, CDK 2 and PCNA, the levels of cyclin B1 and CDC 2 proteins were not changed after GL331 treatment; however, the ability to form complex between cyclin B1 and CDC 2 was obviously affected in GL331-treated NPC cells, which associates with the inhibition of cyclin B1/CDC 2 kinase activity elicited by GL331. These data could provide more principal bases for future therapeutic application of this potential anti-cancer agent.     

LOXL4mRNA在膀胱癌中的表达及意义

膀胱癌是泌尿生殖系统最常见的恶性肿瘤,但其发生、发展的机制不清楚.通过采用逆转录聚合酶链反应(RT-PCR)方法检测58例膀胱癌组织、12例时照膀胱组织LOXL4(lysyl oxidase-like protein 4)mRNA的表达及其与临床分期、病理分级的关系.研究发现,58例膀胱癌组织中,LOXIA mRNA阳性表达率为24.1%(14/58),对照膀胱正常组织中LOXL4 mRNA阳性表达率为100%(12/12),膀胱癌组LOXL4 mRNA阳性表达率明显低于对照组(P<0.05).在膀胱癌组织不同临床分期中,Ta～1期阳性表达率为40%(10/25),T2～4期阳性表达率为12.1%(4/33),T2-4期膀胱癌组LOXL4 mRNA阳性表达率低于Ta～1期膀胱癌(P<0.05).不同病理分级膀胱癌组织中,G1阳性表达率为42.9%(9/21),G2～3阳性表达率为13.5%(5,37),G2～3膀胱癌组LOXIA mRNA阳性表达率低于G1膀胱癌(P<0.05).结果表明膀胱癌组LOXL4 mRNA表达水平明显低于正常对照组,LOXL4 mRNA的表达与膀胱癌的临床分期和病理分级呈负相关.提示LOXL4失表达、低表达可能是膀胱癌发生、发展的关键因素之一.     

ARTD1 regulates cyclin E expression and consequently cell-cycle re-entry and G1/S progression in T24 bladder carcinoma cells

ADP-ribosylation is involved in a variety of biological processes, many of which are chromatin-dependent and linked to important functions during the cell cycle. However, any study on ADP-ribosylation and the cell cycle faces the problem that synchronization with chemical agents or by serum starvation and subsequent growth factor addition already activates ADP-ribosylation by itself. Here, we investigated the functional contribution of ARTD1 in cell cycle re-entry and G₁/S cell cycle progression using T24 urinary bladder carcinoma cells, which synchronously re-enter the cell cycle after splitting without any additional stimuli. In synchronized cells, ARTD1 knockdown, but not inhibition of its enzymatic activity, caused specific down-regulation of cyclin E during cell cycle re-entry and G₁/S progression through alterations of the chromatin composition and histone acetylation, but not of other E2F-1 target genes. Although Cdk2 formed a functional complex with the residual cyclin E, p27^{Kip1 Murray AH, Hunt T. The cell cycle: an introduction. New York: Oxford University Press, 1993. [Google Scholar]} protein levels increased in G₁ upon ARTD1 knockdown most likely due to inappropriate cyclin E-Cdk2-induced phosphorylation-dependent degradation, leading to decelerated G₁/S progression. These results provide evidence that ARTD1 regulates cell cycle re-entry and G₁/S progression via cyclin E expression and p27^{Kip1 Murray AH, Hunt T. The cell cycle: an introduction. New York: Oxford University Press, 1993. [Google Scholar]} stability independently of its enzymatic activity, uncovering a novel cell cycle regulatory mechanism.     

Prognostic implication of CDC25A and cyclin E expression on primary breast cancer patients

Defect in cell cycle control is a hallmark character of cancer. We have investigated the association of Ki67 labeling index, cyclin E and CDC25A expressions with clinical follow-up data in breast carcinomas. Flow cytometry was used to detect gene amplification of cyclins in 44 tumor tissue with invasive breast carcinomas. Multivariate Cox proportional hazard ratio test was used to show the correlations. Cyclin E or CDC25A were upregulated in 34% of the tumors. Among the whole total material, expression of cyclin E and of CDC25A were found upregulated in 31.9% and 39.4% of cells, respectively. Both CDC25A and cyclin E protein expression levels were correlated with Ki67 expression level (p < 0.001). In addition, the expression of CDC25A was associated significantly with poor survival (P = 0.028), whereas no correlation was found with cyclin E. These findings suggest a possible prognostic value for CDC25A as a cell cycle marker and may imply in characteristic of high risk breast cancer patients.     

Increased cyclin E expression may obviate the role of cyclin D1 during brain development in cyclin D1 knockout mice

Cyclins D and E play critical roles during the G1 phase of mammalian cell division. Cyclin D1 expression is high and expected to play an important role during mouse brain development. However, in the present study, we found no difference in CNS morphology between cyclin D1 knockout (KO) and control wild-type mice at the ages of 1, 4 and 12 months. Analysis of protein expression in embryonic brains revealed that cyclin E is obviously increased in cyclin D1 KO mice at 13.5 days post coitum. At the same age a high level of cyclin D1 expression is detected in the embryonic brain of wild-type mice. The data indicate that enhanced cyclin E protein expression in cyclin D1 KO mice may obviate the role of cyclin D1 and contribute to the normal brain development of cyclin D1 KO mice.     

The effect of lead on cyclin expression in lupine roots

The expression of cell cycle gene, cyclin, was analyzed in lupine roots exposed to lead. The level of cyclin mRNA and coding protein were examined by Northern and Western blot techniques and by using lupine cDNA (CycB1;1) and animal-derived antibody, respectively. The cyclin mRNA level was either unchanged or slightly increased after lead treatment and it was concomitant with decrease of RNase activity. The lead ions caused the decrease of cyclin protein accumulation and this effect may be responsible for previously described reduction of mitotic activity caused by this heavy metal (Przymusiński and Woźny 1985).