脑胶质瘤中p14ARF、MDM2及p53的表达及其意义

目的：探讨p14^ARF、MDM2和p53蛋白在脑胶质瘤组织中的表达及p14^ARF-MDM2-p53通路在胶质瘤进展中的意义。方法：应用免疫组化SP法检测25例Ⅱ级和23例Ⅳ级胶质瘤组织中-p14^ARF、MDM2和-p53蛋白的表达，并分析其与胶质瘤组织学分级之间的关系。结果：Ⅱ级和Ⅳ级胶质瘤中，MDM2蛋白的阳性表达率分别为24．00％（6／25）及56．52％（13／23）（P=0．021），p53蛋白的阳性表达率分别为28．00％（7／25）及60．87％（14／23）（P=0．022），二者阳性表达率均随胶质瘤恶性程度的增加而升高，Spearman等级相关分析显示，MDM2、p53的表达与胶质瘤分级呈正相关；p14^ARF的阳性表达率分别为76．00％（19／25）及34．78％（8／23）（P=0．004），其阳性表达率随胶质瘤恶性程度的增加而降低，Spearman等级相关分析显示，p14^ARF的表达与胶质瘤分级呈负相关（P〈0．05）。结论：脑胶质瘤组织中，MDM2和p53均呈不同程度的过度表达，且随恶性程度的增加表达水平增高；而p14^ARF随恶性程度的增加而表达水平降低。MDM2扩增、p53突变及p14^ARF蛋白低表达均与胶质瘤的进展有关。     

脑胶质瘤中p14~(ARF)、MDM2及p53的表达及其意义

目的:探讨p14ARF、MDM2和p53蛋白在脑胶质瘤组织中的表达及p14ARF-MDM2-p53通路在胶质瘤进展中的意义。方法:应用免疫组化SP法检测25例Ⅱ级和23例Ⅳ级胶质瘤组织中-p14ARF、MDM2和-p53蛋白的表达,并分析其与胶质瘤组织学分级之间的关系。结果:Ⅱ级和Ⅳ级胶质瘤中,MDM2蛋白的阳性表达率分别为24.00%(6/25)及56.52%(13/23)(P=0.021),p53蛋白的阳性表达率分别为28.00%(7/25)及60.87%(14/23)(P=0.022),二者阳性表达率均随胶质瘤恶性程度的增加而升高,Spearman等级相关分析显示,MDM2、p53的表达与胶质瘤分级呈正相关;p14ARF的阳性表达率分别为76.00%(19/25)及34.78%(8/23)(P=0.004),其阳性表达率随胶质瘤恶性程度的增加而降低,Spearman等级相关分析显示,p14ARF的表达与胶质瘤分级呈负相关(P<0.05)。结论:脑胶质瘤组织中,MDM2和p53均呈不同程度的过度表达,且随恶性程度的增加表达水平增高;而p14ARF随恶性程度的增加而表达水平降低。MDM2扩增、p53突变及p14ARF蛋白低表达均与胶质瘤的进展有关。     

人脑胶质瘤p53基因突变及MDM2、p16、p53蛋白表达与临床病理特征相关性研究

目的:探讨p53突变在人脑胶质瘤发生发展中的作用及p53蛋白蓄积与突变的符合程度,并研究MDM2、p16、p53蛋白表达与胶质瘤临床病理特征的关系以及三者的相关性。方法:利用聚合酶链反应-单链构象多态性分析法(PCR-SSCP)及LSAB免疫组化法对已明确诊断的48例人脑胶质瘤进行p53基因突变以及MDM2、p16、p53蛋白表达的检测。结果:48例胶质瘤中20例p53蛋白呈阳性表达(41.7%)。PCR-SSCP检测发现17例(35.4%)呈现p53基因的单链构象多态性改变,均位于5～8外显子,突变例数依次为7(41.2%)、1(5.9%)、4(23.5%)、5(29.4%)。两种方法检测的符合率为89.6%(43/48)。MDM2、p53蛋白阳性率分别为22.9%、41.7%,p16表达缺失率为60.4%。在高级别(Ⅲ、Ⅳ级)的肿瘤中p53表达率及p16表达缺失率分别为63.2%、84.2%,均明显高于低级别(Ⅱ级)的肿瘤(分别为27.6%、44.8%)(P<0.05)。在不同分级的胶质瘤中,MDM2表达率及阳性程度没有显著差异。在p53阳性表达的病例中常伴有p16的表达缺失(57.6%),而且大多出现在Ⅲ、Ⅳ级肿瘤中(9/12)。p53突变、三种分子的表达均与患者的年龄、性别、肿瘤的大小及发病部位无相关性。结论:胶质瘤中p53基因的突变与胶质瘤的发生及恶性进展相关,p53蛋白蓄积与突变率较一致。MDM2基因异常参与胶质瘤的发生,是其形成的早期事件;p16、p53在胶质瘤中的异常表达与肿瘤的分化程度相关。     

MDM2、P53蛋白在星形细胞瘤中的表达

目的　探讨 MDM2、p5 3蛋白在星形细胞瘤发病中的作用及其与肿瘤病理分级、预后的关系。方法　对确诊并有随访的 36例星形细胞瘤标本用免疫组化技术 SABC法进行 MDM2、p5 3蛋白定位观察。结果　发现 MDM2及 p5 3蛋白表达阳性率分别为 44 .4% (16 / 36例 )和 38.9% (14/ 36例 ) ,其阳性率与年龄、性别无关 ,但与肿瘤病理分级呈显著正相关 (分别为 P<0 .0 5 ,P<0 .0 1)。在 36例星形细胞瘤中MDM2和 p5 3共同阳性表达 9例 (2 5 % ) ,这 9例与单独MDM2蛋白或 p5 3蛋白阳性表达病例及 MDM2和 p5 3蛋白表达均阴性病例的一年和三年存活率有显著差别 (分别为P<0 .0 5 ,P<0 .0 1)。结论　 MDM2、p5 3蛋白阳性检出率有助于判断星形细胞瘤的恶性程度及预测肿瘤的预后     

bcl-2蛋白表达在脑胶质瘤中的意义

目的:研究人脑胶质瘤中bcl-2蛋白表达强度与胶质瘤的临床病理分级及患者预后间的关系。方法:应用免疫组化技术检测58例脑胶质瘤患者手术切除标本中bcl-2蛋白表达强度、HE染色确定肿瘤病理分级、门诊及电话随访,掌握患者术后生存时间。结果:bcl-2蛋白表达强度与肿瘤病理分级相关(P<0.01)、与患者术后生存时间相关(P<0.01)。bcl-2蛋白表达愈强,胶质瘤临床病理分级愈高、患者术后生存时间也愈短。结论:bcl-2蛋白表达水平与胶质瘤恶性程度成正比、与胶质瘤患者术后生存时间成反比,是评价胶质瘤恶性程度和预后的客观指标之一。     

MDM2与MMP-9在骨肉瘤组织中的表达及临床意义

目的探讨骨肉瘤中MDM2、MMP-9的表达与骨肉瘤病理特征、转移及预后的关系。方法 采用免疫组织化学(S-P)法检测61例骨肉瘤标本中MDM2、MMP-9的表达情况, 并分析MDM2、MMP-9的表达与骨肉瘤病理特征、转移及预后的关联性。同时设20例正常骨组织为阴性对照组。结果 MDM2、MMP-9在骨肉瘤组织中的阳性表达率均高于正常骨组织(P＜0.001), 其表达与性别、年龄及病理分级无关(P＞0.05), 与骨肉瘤的转移和预后相关(P＜0.05), 且二者表达呈正相关(P＜0.05)。二者同时阳性表达与骨肉瘤转移有高度关联性, 其生存率明显低于同时阴性表达者, 有显著性差异(P＜0.001)。结论 MDM2、MMP-9表达异常的骨肉瘤组织恶性程度高、预后差。MDM2、MMP-9蛋白表达的改变可能与骨肉瘤发生发展相关, 检测骨肉瘤中MDM2和MMP-9的表达水平对其预后评估有重要意义。     

MDM2和p53在反应性及肿瘤性星形胶质细胞中的表达

目的：检测MDM2和p53蛋白在星形胶质细胞反应性增生与星形胶质细胞瘤中的表达，探讨二者在胶质瘤形成和发展中的作用及其相关性。方法：应用组织芯片和免疫组化染色技术检测正常脑组织、星形胶质细胞反应性增生、低级别（Ⅰ-Ⅱ级）和高级别（Ⅲ-Ⅳ级）星形胶质细胞瘤中MDM2和p53蛋白的表达情况。结果：正常脑组织中MDM2和p53蛋白均呈阴性表达；反应性增生组、低级别肿瘤组及高级别肿瘤组中MDM2蛋白的阳性率分别为32.7％（16／49）、59．2％（29／49）、80．0％（40／50）；p53蛋白的阳性率分别为27．3％（12／49）、57．1％（28／49）、82．0％（41／50）。二者阳性表达率均随着病变恶性程度的增加而升高，MDM2和p53在各实验组间的比较差异均有统计学意义（P〈0．05）；且MDM2和p53表达密切相关（P〈0．05）。结论：MDM2和p53在星形胶质细胞反应性增生及星形胶质细胞瘤中均呈不同程度的过度表达，且随着病变恶性程度的增加表达水平增高，MDM2扩增和p53突变是胶质瘤发生的早期事件，二者的联合检测可能会对星形胶质细胞反应性增生与低级别胶质细胞瘤的鉴别诊断以及星形胶质细胞瘤的早期诊断提供一定的依据。     

p16mRNA和MDM2mRNA在恶性纤维组织细胞瘤中的表达及意义

[目的]探讨p16mRNA和MDM2mRNA在恶性纤维组织细胞瘤组织中的表达及其与肿瘤浸润、转移的关系，分析两者表达的临床意义。[方法]应用原位杂交方法检测66例恶性纤维组织细胞瘤组织中P16mRNA、MDM2mRNA的表达。[结果]P16mRNA和MDM2mRNA在肿瘤组织中的阳性表达率分别为48.5％和66.7％；P16mRNA和MDM2mRNA的阳性表达率与临床分期、病理分化程度和生存期显著性相关（P〈0.05）；而与患者的性别、年龄和组织学类型无关（P〉0.05）。[结论]p16和MDM2在恶性纤维组织细胞瘤发生、发展中起重要作用,而且可能是预测恶性纤维组织细胞瘤预后的生物学指标。     

原发性胶质母细胞瘤和继发性胶质母细胞瘤中MMP-2蛋白及MDM2蛋白的表达和意义

目的:探讨MMP-2蛋白及MDM2蛋白在原发性胶质母细胞瘤和继发性胶质母细胞瘤中的表达差异性及其意义。方法:免疫组化SP二步法检测72例胶质母细胞瘤（36例原发性胶质母细胞瘤和36例继发性胶质母细胞瘤）石蜡包埋标本及10例正常脑组织石蜡包埋标本MMP-2蛋白和MDM2蛋白的表达情况,分析上述蛋白在两种病理类型的胶质母细胞瘤中的表达差异情况。结果:在原发性和继发性胶质瘤标本中,MMP-2蛋白的阳性表达率分别为63.9%和86.1%;MDM2蛋白的阳性表达率分别为61.1%和80.6%。MMP-2蛋白及MDM2蛋白在原发性和继发性胶质母细胞瘤中的表达差异存在统计学意义（P＜0.05）。结论:MMP-2蛋白及MDM2蛋白在原发性、继发性胶质母细胞瘤中的表达差异可反映它们的不同分子遗传学特征。     

胶质瘤p27Kip1、bcl-2和PCNA蛋白的表达

目的　探讨胶质瘤p2 7Kip1,bcl 2和PCNA蛋白表达与肿瘤恶性程度、细胞增殖活性、凋亡程度的关系。方法　采用免疫组化染色S P法检测 66例不同级别的胶质瘤 p2 7Kip1,bcl 2和PCNA蛋白的表达。结果　在 66例胶质瘤中 ,p2 7Kip1表达 18例(2 7% ) ,bcl 2表达 2 0例 (3 0 % ) ,PCNA表达 5 1例 (77% )。p2 7Kip1蛋白表达率随着胶质瘤级别升高而减少 ;bcl 2蛋白表达率随肿瘤级别升高而相应增加 ;PCNA表达随胶质瘤级别升高阳性反应强度增加 ;但Ⅰ、Ⅱ与Ⅲ级和Ⅳ级组间无显著性差异。结论　p2 7Kip1蛋白表达的缺失可能与胶质瘤的发生有关 ;bcl 2基因可能间接抑制细胞凋亡而与胶质瘤的分型、细胞的增殖活性以及潜在的临床行为无直接关系 ;PCNA的表达与星形胶质细胞瘤的恶性行为有关     

Effects of cytotoxicity of 2-chloro-2'-deoxyadenosine and 2-bromo-2'-deoxyadenosine on cell growth, clonogenicity, DNA synthesis, and cell cycle kinetics

The cytotoxic effects of the adenosine deaminase resistant analogues 2-bromo-2'-deoxyadenosine (2-BrdAdo) and 2-chloro-2'-deoxyadenosine (2-CldAdo) have been compared with those of deoxyadenosine (dAdo). Like 2-CldAdo, 2-BrdAdo is highly effective in inhibiting the growth of many T-lymphoblastoid, B-lymphoblastoid, and myeloid cell lines in culture. Concentrations required to inhibit growth of CCRF-CEM human T-lymphoblastoid cells by 50% (IC50) are: 2-CldAdo, 0.045 microM; 2-BrdAdo, 0.068 microM; dAdo, 0.9 microM in the presence of 5 microM erythro-9-(2-hydroxy-3-nonyl)adenine. Like dAdo, 2-BrdAdo causes a much greater decrease in DNA synthesis than in RNA and protein synthesis. For each of the nucleosides the concentration required to cause 50% inhibition of DNA synthesis (as measured by thymidine incorporation) in an 18-h exposure is very similar to the IC50 for growth and to the concentration required to decrease viability (clonogenicity) over 18 h by 50% (EC50). A fraction of CCRF-CEM cells (approximately equal to 30%) is resistant to killing by exposure to 2-BrdAdo or 2-CldAdo for 4 h at concentrations 100 times the EC50, but 3% of cells are resistant to exposure for 4 h to a concentration of dAdo 3 times the EC50. Each of the three nucleosides causes accumulation of cells in S phase, the accumulation becoming more marked with longer periods of exposure and with higher concentrations of nucleoside. During exposures for 18-24 h at a concentration of nucleoside near the EC50 most cells accumulate in S, with most in early S, whereas exposure to concentrations greater than EC95 accumulates cells at the G1/S border. This suggests that loss of viability is associated with a blockade of some process specifically occurring at the initiation of S phase. At an optimum dosage schedule, 2-BrdAdo and 2-CldAdo have similar therapeutic effects against L1210 in vivo, both producing over 99% cell kill, but the optimum dosage of 2-CldAdo is lower.     

Expression of heparanase, Mdm2, and erbB2 in ovarian cancer

Ovarian cancer is the most lethal of gynecological malignancies. Yet early diagnosis and prognosis are far from being satisfactory. Degradation of heparan sulfate proteoglycans by heparanase appears to play an important role in the invasiveness of tumor cells through the basement membrane and into the extracellular matrix. Recent cloning of the heparanase gene and generation of monoclonal antibodies against the enzyme permit to examine tumor cell expression of the enzyme. The aim of the present study was to assess heparanase activity and localization in various subtypes of epithelial ovarian cancer in correlation with oncogene expression. Histologically confirmed malignant ovarian tissue from ten women and tissue from 2 benign ovarian tumors and 4 normal ovaries were assessed for heparanase presence, activity and localization, incidence of apoptosis and expression of the oncogenes erbB2 and Mdm2. Heparanase immunohistostaining and activity were present in mucinous carcinomas and were more intense than in endometrioid and in serous carcinomas. The lowest activity was observed in benign ovarian tumors and normal ovaries. In ovarian carcinomas the enzyme was intensely concentrated in the cytoplasm of the cancerous cells. In contrast, in normal ovaries and benign tumors the enzyme was predominantly localized in endothelial cells lining blood capillaries. The rate of apoptosis was considerably higher in mucinous and endometrioid carcinomas, and was lower in serous and primary peritoneal carcinomas. Extremely high concentration of heparanase was often demonstrated in apoptotic cells. Endometrioid and serous carcinomas showed high expression of Mdm2 and erbB2 while mucinous carcinomas showed low expression. In benign ovarian tumors and normal ovaries the expression of both oncoproteins was extremely low. In conclusion ovarian carcinomas demonstrate higher levels of heparanase than benign tumors and normal ovaries suggesting that the enzyme may play an important role in metastatic spread of the cancerous cells. Apoptosis may be a significant part of the mechanism of the enzyme release into the extracellular space. Although heparanase activity seems to play an essential role in tumor progression, expression of oncogenes, such as erbB2 and Mdm2 seems to play the dominant role in the development of ovarian cancer.     

1,1-双(4-氯苯)-2,2,2-三氯乙烷对大鼠神经系统的氧化应激作用

背景与目的：研究1,1-双(4-氯苯)-2,2,2-三氯乙烷（DDT）对大鼠大脑皮层、海马和小脑组织诱导的氧化应激作用。 材料与方法：SD大鼠共24只, 随机分为4组, 即溶剂对照组与低 （20mg/kg）、中 （40mg/kg）、高（80mg/kg）3个DDT剂量组,每组6只,大鼠经口灌胃染毒7 d后断头处死,测定大脑皮层、海马和小脑组织丙二醛（MDA）水平和总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-PX）和谷胱甘肽还原酶(GR)活力。 结果：①与溶剂对照组比较,随着DDT浓度的增加,大鼠大脑皮层、海马和小脑中MDA含量升高,T-SOD活性下降;② 大脑皮层GSH-PX活性在低剂量时升高,在中、高剂量组则显著下降,海马GSH-PX活性随着剂量的增加而显著下降,而小脑GSH-PX活性则在中、高剂量组出现显著下降;③ 海马GR活性在中、高剂量组随着剂量的增加而显著下降,在小脑则只有高剂量组出现了下降,皮层GR活性在各剂量组未观察到明显改变。结论：DDT可以引发神经组织的脂质过氧化反应增强。DDT导致机体组织的氧化损伤可能在动物神经毒性中起重要作用。     

Associations with growth factor genes (FGF1, FGF2, PDGFB, FGFR2, NRG2, EGF, ERBB2) with breast cancer risk and survival: the Breast Cancer Health Disparities Study

Growth factors (GF) stimulate cell proliferation through binding to cell membrane receptors and are thought to be involved in cancer risk and survival. We examined how genetic variation in epidermal growth factor (EGF), neuregulin 2 (NRG2), ERBB2 (HER2/neu), fibroblast growth factors 1 and 2 (FGF1 and FGF2) and its receptor 2 (FGFR2), and platelet-derived growth factor B (PDGFB) independently and collectively influence breast cancer risk and survival. We analyzed data from the Breast Cancer Health Disparities Study which includes Hispanic (2,111 cases, 2,597 controls) and non-Hispanic white (1,481 cases, 1,586 controls) women. Adaptive rank-truncated product (ARTP) analysis was conducted to determine gene significance. Odds ratios (OR) and 95 % confidence intervals were obtained from conditional logistic regression models to estimate breast cancer risk and Cox proportional hazard models were used to estimate hazard ratios (HR) of dying from breast cancer. We assessed Native American (NA) ancestry using 104 ancestry informative markers. We observed few significant associations with breast cancer risk overall or by menopausal status other than for FGFR2 rs2981582. This SNP was significantly associated with ER+/PR+ (OR 1.66, 95 % CI 1.37–2.00) and ER+/PR? (OR 1.54, 95 % CI 1.03–2.31) tumors. Multiple SNPs in FGF1, FGF2, and NRG2 significantly interacted with multiple SNPs in EGFR, ERBB2, FGFR2, and PDGFB, suggesting that breast cancer risk is dependent on the collective effects of genetic variants in other GFs. Both FGF1 and ERBB2 significantly influenced overall survival, especially among women with low levels of NA ancestry (P _ARTP = 0.007 and 0.003, respectively). Our findings suggest that genetic variants in growth factors signaling appear to influence breast cancer risk through their combined effects. Genetic variation in ERBB2 and FGF1 appear to be associated with survival after diagnosis with breast cancer.