9-顺维A酸诱导肺癌细胞L78凋亡及其与Rb和Cyclin D_1基因表达的关系

目的 研究 9-顺维 A酸 (9- cis- retinoic acid,9- cis RA)诱导肺鳞癌细胞株 L 78凋亡作用及其与 Rb、Cy-clin D1 基因表达的关系 ,初步探讨其作用机制。 方法 体外培养肺鳞癌细胞株 L78,随机分为两组 ,实验组 :加入浓度为 5 μmol/ L 的 9- cis RA;对照组 :加入浓度为 0 .1%二甲亚砜。两组均培养 4 8小时后用免疫组织化学法检测 L78细胞中 Rb基因表达率 ,用流式细胞仪技术分别检测 Rb、Cyclin D1 基因表达率和肿瘤细胞凋亡率 ,并研究三者之间的相关关系。 结果 实验组肺鳞癌细胞株 L78细胞 Rb基因表达明显增强 ,Cyclin D1 基因表达降低 ,细胞凋亡率增高(P<0 .0 1)。 Rb基因表达率与细胞凋亡发生率呈正相关 (r=0 .85 4 ,P<0 .0 5 ) ,Cyclin D1 基因表达率与细胞凋亡发生率呈负相关 (r=- 0 .812 ,P<0 .0 5 )。 结论 9- cis RA可能通过 Rb基因表达增强和 Cyclin D1 基因表达降低途径 ,使细胞明显阻滞在 G0 / G1 期 ,并诱导肺鳞癌细胞株 L 78细胞凋亡。

P16、Cyclin D_1蛋白表达与胰腺癌生物学行为

目的 探讨 P16、Cyclin D1 蛋白表达水平与胰腺癌发生、发展及预后的关系。 方法 应用免疫组织化学 SP法结合微波抗原修复法对 5 3例原发性胰腺癌和 4 2例胰腺良性病变及正常组织中 P16蛋白及 Cyclin D1蛋白进行检测。 结果 胰腺癌组织、胰腺良性病变和正常组织中 P16蛋白的阳性率分别为 6 6 .0 4 %和 85 .71% (P<0 .0 5 ) ;Cyclin D1 表达的阳性率分别为 5 2 .83%和 2 3.81% (P<0 .0 5 ) ,P16及 Cyclin D1 蛋白表达与胰腺癌的组织分化、病理分期及术后生存期密切相关。 结论 P16及 Cyclin D1

Cyclin D_1、p27在肺癌中表达的研究

目的 研究 Cyclin D1 、p2 7基因蛋白在肺癌组织中的表达与肺癌临床病理特征的关系。方法 应用免疫组化 S- P法检测 10 9例手术切除肺癌组织标本中两种基因蛋白的表达。结果 肺癌 Cyclin D1 基因过度表达的总阳性率为 93.6 % (10 2 / 10 9)。其中鳞癌为 98% (5 0 / 5 1) ,腺癌为 97.2 % (35 / 36 ) ,小细胞癌为 77.3% (17/ 2 2 ) ;p2 7基因表达的阳性率为 33% (36 / 10 9) ,其中鳞癌为 39.1% (2 0 / 5 1) ,腺癌为 2 5 % (9/ 36 ) ,小细胞癌 31.8% (7/ 2 2 )。CyclinD1 阳性表达与肺癌的分化程度、TNM分期呈负相关。Cyclin D1 和 p2 7蛋白在肺癌组织中的表达呈负相关。结论 Cyclin D1 和 p2 7基因蛋白表达与肺癌发生。

CyclinD_1和ras过表达在胃粘膜细胞癌变中的协同作用

目的 探讨cyclinD1和ras在胃癌中的相互关系。方法 运用免疫组化方法检测cyclinD1和ras在胃癌中的表达。结果 cyclinD1和ras在胃癌中的阳性表达率分别为 59 2 %和 57 1% ,二者在胃癌中的表达显著相关 (P <0 0 5)。cyclinD1阳性表达率与肿瘤大小和组织学类型显著相关 (P<0 0 5)。ras过表达与分化程度和Lauren分型显著相关 (P <0 0 5)。二者过表达与胃癌的淋巴结状况、浸润深度和临床病理分期无关。结论 cyclinD1和ras过表达在胃粘膜细胞癌变中具有协同作用。

口腔鳞癌组织中Cyclin D_1、Ki-67基因的表达及意义

目的 探讨口腔鳞癌组织中 Cyclin D1 、Ki- 6 7基因蛋白表达与口腔鳞癌发生、发展的关系。方法 取 6 8份口腔鳞癌组织 ,用免疫组化法检测癌组织中 Cyclin D1 、Ki- 6 7基因蛋白的表达 ,用积分 PCR方法检测Cyclin D1 的基因扩增。结果 口腔鳞癌组织中 Cyclin D1 基因蛋白表达阳性 36例 (5 2 .9% ) ,Ki- 6 7基因蛋白表达阳性 4 8例 (70 .5 % ) ,Cyclin D1 与 Ki- 6 7基因蛋白表达呈正相关 (r=0 .786 1) ,33份 (4 8.5 % )口腔鳞癌组织中Cyclin D1 基因存在扩增 ,Cyclin D1 基因扩增与 Cyclin D1 基因蛋白表达无相关性。Cyclin D1 基因蛋白的表达与淋巴结转移有关 ,Ki- 6 7基因蛋白的表达与肿瘤分化程度有关。结论 Cyclin D1 基因扩增及 Ki- 6

Prognostic value of MET, cyclin D1 and MET gene copy number in non-small cell lung cancer

The aim of this study was to analyze the correlation of the expression of MET and cyclin D1 and MET gene copy number in non-small cell lung cancer （NSCLC） tissues and patient clinicopathologic characteristics and sur- vival. Sixty-one NSCLC tissue specimens were included in the study. The expression of MET and cyclin D1 was evaluated by immunohistochemistry and MET gene copy number was assessed by quantitative real-time polymer- ase chain reaction （Q-PCR）. Positive expression of MET and cyclin D1 protein and increased MET gene copy number occurred in 59.0%, 59.0% and 18.0% of 61 NSCLC tissues, respectively. MET-positivity correlated with poor differentiation （P = 0.009）. Increased MET gene copy number was significantly associated with lymph node metastasis （P = 0.004） and advanced tumor stage （P = 0.048）, while the expression of cyclin D1 was not associ- ated with any clinicopathologic parameters. There was a significant correlation between the expression of MET and MET gene copy number （P = 0.002）. Additionally, the expression of cyclin D1 had a significant association with the expression of MET as well as MET gene copy number （P = 0.002 and P = 0.017, respectively）. MET- positivity and increased MET gene copy number were significantly associated with poor overall survival （P = 0.003 and P 〈 0.001, respectively） in univariate analysis. Multivariate Cox proportional hazard analysis confirmed that the expression of MET and MET gene copy number were prognostic indicators of NSCLC （P = 0.003 and P = 0.001, respectively）. The overexpression of MET and the increased MET gene copy number might be adverse prognostic factors for NSCLC patients. The activation of the MET/cyclin D1 signaling pathway may contribute to carcino- genesis and the development of NSCLC, and may represent a target for therapy.

Antisense to cyclin D1 reverses the transformed phenotype of human gastric cancer cells

AIM To further investigate the effect of cyclin D1 on the biologic behavior of cancer cells and its potential role in gene therapy of tumor. METHODS A cyclin D1 subcloning plasmid termed BKSD1 was constructed by subcloning the human cyclin D1 cDNA into Bluescript KS, a plasmid vector with a pair of T7 and T3 promoters, with recombinant DNA technology of molecular biology. So, it is easy to generate digoxigenin (DIG) labeled RNA probes of antisense and sense to cyclin D1 using RKSD1 as a template vector. PDORD1AS, an eukaryotic expression vector containing the full length human cyclin D1 cDNA in its antisense orientation cloned into the retroviral vector pDOR neo, was successfully constructed with BKSD1 to change restriction sites. A gastric cancer cell line, SGC7901/VCR, was transfected with pDORD1AS by Lipofect Amine mediated introduction and a subline termed SGC7901/VCRD1AS, which had stable overexpression of antisense RNA to cyclin D1, was obtained by selection in G418. The subline, control subline transfected pDOR neo and SGC7901/VCR were evaluated by methods of immunohistochemistry, flow cytometry, molecular hybridization, morphology and cell biology. RESULTS Compared with control cell lines, SGC7901/VCRD1AS had a reduced expression of cyclin D1 (inhibition rate was about 36%), increased cell size and cytoplasm to nucleus ratio, increased doubling time (42 2h to 26 8h and 26 4h), decreased saturation density (18 9×10 4 to 4 8×10 5 and 4 8×10 5), increased percentage of cells in the G1/G0 phase (80 9%-64 6% and 63 8%), reacquired serum dependence, and a loss of tumorigenicity in nude mice (0/4 to 4/4 and 4/4). CONCLUSION Stable overexpression of antisense RNA to cyclin D1 can reverse the transformed phenotype of human gastric cancer cells and may provide an approach of gene therapy for gastric cancer.

Transretinoic acid inhibits rats gastric epithelial dysplasia induced by N-methyi-N-nitro-N-nitrosoguanidine:influences on cell apoptosis and expression of its regulatory genes

INTRODUCTIONGastric epithelial dysplasia (GED) hypothetically is a straight-forward concept: dysplastic epithelium replacing the normal gastric epithelium of the stomach [1].In the stomach ,like any other segment of the gut ,it is defined as an unequivocal non-invasive epithelial change[2,3].The observation of gastric dysplasia as a cancerous lesion was recognized over a century ago ,but it is only after the advent of gastroscopy that its clinical significance has been stressed[4-7].

细胞周期素D_1在结肠癌中的表达

目的 探讨细胞周期素 ( cyclin) D1与 p53基因在结肠癌中的表达意义及其关系。 方法 应用原位杂交技术和免疫组织化学技术检测结肠癌中 cyclin D1基因 m RNA和 p53蛋白的表达。统计处理采用卡方检验。 结果 发现结肠癌组织和癌旁组织中 cyclin D1表达阳性率分别为 63 .4 % ( 40 / 63 )及 4 4 .4 4 % ( 2 8/ 63 ) ,两者之间差异有显著性 ( P<0 .0 5)。cyclin D1表达程度与癌组织分化程度及临床分期有关 ,差异有显著性 ( P<0 .0 5)。cyclin D1表达与其他临床病理指标无关 ( P>0 .0 5)。p53蛋白在结肠癌中阳性率为 3 8.0 9% ( 2 4 / 63 ) ,p53阳性的患者其 cyclinD1的表达较之 p53阴性者为高 ,差异有显著性 ( P<0 .0 5)。 结论 cyclin D1的过表达及 p53突变在结肠癌的癌变中可能起重要作用 ,cyclin D1的表达参与了癌细胞分化及发展过程。

Pin1和周期素D1在胰腺癌中的表达及其意义

目的检测胰腺癌及癌旁组织中Pin1和周期素D1基因的表达,探讨Pin1在胰腺癌发病中所起的作用。方法收集27例胰腺肿瘤组织及其相应的肿瘤旁组织标本,采用实时荧光定量逆转录聚合酶链反应法(RQ RT-PCR)检测胰腺良恶性肿瘤及肿瘤旁组织中Pin1和周期素D1 mRNA的表达,运用Fisher精确概率分析两者之间的相关性及其与肿瘤临床分期和病理特征的关系。结果7例胰腺囊腺瘤中周期素D1和Pin1的表达与肿瘤旁组织之间无显著性差异;而20例胰腺癌中周期素D1和Pin1的表达明显高于肿瘤旁组织[(2.78±1.02)vs.(4.36±1.27)和(5.48±1.69)vs.(9.97±1.86),P<0.05)]。Pin1和周期素D1与肿瘤的临床分期和病理分化程度无明显的相关(组间差异均为P>0.05),但Pin1与周期素D1的表达有关(P<0.01)。结论胰腺癌中Pin1的过表达可促进周期素D1表达,由此诱导了肿瘤的发生;Pin1可能在胰腺癌中起着重要作用。

BCL-1 Rearrangement in Acute Lymphocytic Leukemia and Its Clinical Significance

BCL 1 rearrangement (BCL 1/IgH gene rearrangement) in acute lymphocytic leukemia and its clinical significance was investigated. In 38 patients with acute lymphocytic leukemia (ALL), the genomic DNA of mononuclear cells isolated from peripheral blood and bone marrow was amplified by using hemi nested polymerase chain reaction (PCR) technique and the expression of cycline D1 protein of mononuclear cells was detected by using immunohistochemical method. Ten patients with acute granulocytic leukemia, 2 with chronic granulocytic leukemia and 10 with normal bone marrow served as control group. The results showed that BCL 1 rearrangement was detectable in 3 of 38 ALL patients (7.9 %) and cyclin D1 protein positive expression was detected in 4 ALL patients (10 5 %). Three ALL patients with BCL 1 rearrangement were all B cell leukemia (B ALL) and accompanied by cyclin D1 protein expression. No BCL 1/IgH rearrangement or cyclin D1 protein expression was detected in 12 patients with granulocytic leukemia and 10 cases of normal bone marrow. Leukocyte counts in peripheral blood of B ALL patients with BCL 1 rearrangement and(or) cyclin D1 protein expression were significantly increased and the patients had bad reaction to chemotherapy. It was concluded that: 1) BCL 1/IgH gene rearrangement were detected in acute B lymphocytic leukemia; 2) B ALL patients with BCL 1 rearrangement and(or) cyclin D1 protein expression had poor prognosis.

Cyclin D_1与肿瘤

CyclinD1为G1期细胞周期蛋白,调节细胞的增殖周期,其表达与肿瘤的发生、发展及预后密切相关。

Expression of cyclin genes in human gastric cancer and in first degree relatives

OBJECTIVE: To clarify the role of these cyclins in human gastric cancer. METHODS: 38 gastric cancer patients, 29 first degree relatives of gastric cancer patients, as well as 18 healthy subjects were included. The mRNA expression of cyclins D1, D2, D3 and E in gastric biopsies was evaluated by RT-PCR analysis using specific primers. Histomorphological features such as intestinal metaplasia, atrophy, H. pylori infection and severity of gastritis were determined by the updated Sydney System. RESULTS: Significant mRNA overexpression was found for cyclins D2, D3 and E compared with healthy normal specimen, but cyclin D1 expression was not different between tumor and normal tissues. In addition, cyclin D2 and D3 overexpression was significantly more frequent in first degree relatives than in healthy controls (P

RhoC反义基因转染对人胆管癌QBC939细胞生长的抑制作用

目的 构建人 Rho C基因反义真核表达载体 ,研究 Rho C基因对胆管癌 QBC939细胞株增殖的影响。 方法 应用分子克隆技术 ,将含 Rho C基因全长开放阅读框目的片段克隆到真核载体 pc DNA 3.1/ Hyg(+)上 ,构建反义 Rho C c DNA真核表达载体 ,以脂质体转染人胆管癌 QBC939细胞。检测细胞生长曲线 ,RT- PCR检测Cyclin D1 在 QBC939细胞、转染空载体的 QBC939细胞 (QBC939- V)和转染反义 Rho C重组载体的 QBC939细胞(QBC939- AS)的 m RNA相对表达量。 结果 与 QBC939细胞及 QBC939- V细胞相比 ,QBC939- AS细胞体外生长较慢。 QBC939- AS细胞、QBC939- V细胞和 QBC939细胞的 Cyclin D1 的表达量分别为 0 .15 1± 0 .0 76 ,0 .2 96±0 .0 2 6和 0 .2 87± 0 .0 5 7;QBC939- AS细胞的 Cyclin D1 的表达量与 QBC939- V细胞及 QBC939细胞的差异有显著性 (P<0 .0 5 )。 结论 Rho C可能通过调节 Cyclin D1 来抑制 QBC939细胞的增殖能力。

抑癌基因PTEN在非小细胞肺癌发生发展中的作用

目的 研究抑癌基因 10q丢失的与张力蛋白同源的磷酸酶基因 (PTEN)在非小细胞肺癌 (NSCLC)组织中的表达水平与NSCLC的发生和发展的关系 ,以及PTEN和抑癌基因p2 7Kip1、细胞周期蛋白D1(CyclinD1)表达的相关性。方法 以免疫组化方法分别检测 6 2例石蜡包埋的NSCLC组织中PTEN、p2 7Kip1和CyclinD1蛋白表达水平 ;用原位杂交方法检测 2 1例新鲜NSCLC组织中PTENmRNA的表达情况。结果 PTEN的表达在肺鳞癌中阳性率为4 0 0 % ( 12 / 30 ) ,肺腺癌中为 4 3 75 % ( 14 / 32 ) ;中高分化组肺癌中阳性率为 5 4 2 9% ( 19/ 35 ) ,低分化组中为2 5 93% ( 7/ 2 7) ;淋巴结转移组阳性率为 4 0 91% ( 9/ 2 2 ) ,无转移组阳性率为 5 4 84 % ( 17/ 31)。 2 1例新鲜肺癌标本原位杂交结果显示 ,鳞癌阳性率为 4 1 6 7% ( 5 / 12 ) ,腺癌阳性率为 33 33% ( 3/ 9)。同时检测了 p2 7Kip1和CyclinD1在肺癌中的表达状况。PTEN与p2 7Kip1表达呈正相关 ( χ2 =4 2 6 6 6 7,P <0 0 5 ) ,与CyclinD1的表达呈负相关 ( χ2 =4 5 5 814 ,P <0 0 5 )。结论 PTEN的失表达在NSCLC的发生、发展和转移过程中扮演着重要的角色。其失表达与NSCLC的分化程度 (P <0 0 5 )、淋巴结转移 (P <0 0 5 )有关 ,而与其组织学类型无关 (P >0

急性髓系白血病患者hDMP1表达特点及临床意义

目的探讨人类细胞周期调节蛋白依赖性Myb样蛋白1(hDMP1)在急性髓系白血病(AML)中的表达及其临床意义。方法 93例初诊AML患者及19例健康志愿者(对照组)骨髓标本,以β胆色原脱氢酶(GAPDH)表达水平为内参,应用实时定量RT-PCR方法检测hDMP1及肾母细胞瘤基因(WT1)的表达水平。分析hDMP1表达水平与患者临床资料、染色体核型的关系。结果AML患者骨髓细胞中hDMP1在表达水平较对照组显著降低(476.6vs.2103.0)(P<0.01),WT1基因表达水平较对照组显著升高(202.6vs.1.2)(P<0.01)。hDMP1和WT1基因表达水平与患者性别、年龄、有无肝脾淋巴结肿大、起病时外周血WBC计数、骨髓中原始细胞比例、AML亚型、染色体核型分组无关。结论 AML患者hDMP1的低表达伴随WT1基因高表达,两者均参与AML发病。

Effect of Artemisia species on cellular proliferation and apoptosis in human breast cancer cells via estrogen receptor-related pathway

OBJECTIVE:To investigate the mechanism underlying the anticancer effect of Artemisia species through the inhibition of cell growth and induction of apoptosis in breast carcinoma cells.METHODS:To evaluate the anticancer activity of methanol extracts of eight Artemisia species(Artemisia stolonifera,Artemisia selengensis,Artemisia japonica,Artemisia Montana,Artemisia capillaris,Artemisia sylvatica,Artemisia keiskeana,and Artemisia scoparia),we first investigated the proliferation of estrogen receptor(ER)-positive MCF-7breast carcinoma cells exposed to 5 or 200 g/mL for72 h.Apoptosis induction was assessed by an Annexin V binding assay in cells exposed to extracts at a high concentration(200 g/mL).To verify the mechanism of apoptosis,ER expression and its related signaling was investigated using an immunoblot assay under the same conditions.RESULTS:MCF-7 cells showed the strongest antiproliferative response to the tested extracts.Howev-er,a biphasic effect was observed:the extracts inhibited proliferation at high concentrations whereas they stimulated it at low ones.ER expression was similarly modulated by the extracts.However,all of the extracts induced apoptosis at a high concentration(200 g/mL).Compared to the control level,exposure to the extracts resulted in a remarkable increase in the shift of cell populations.CONCLUSION:The present study suggests that the tested Artemisia species exerted their anticancer effects through the induction of apoptosis via an ER-related pathway.