胶质瘤p53、bcl-2、MDM2表达与细胞增殖和凋亡的关系

目的：研究胶质瘤中ｐ５３ 、ｂｃｌ２ 、ＭＤＭ２ 表达与细胞增殖和凋亡的关系。方法：对４８ 例胶质瘤用免疫组化法检测ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达;以Ｋｉ６７ 标记指数和ＡｇＮＯＲ 计数检测其增殖活性;用ＴＵＮＥＬ 法检测细胞凋亡。结果：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达阳性率分别为４７－９ ％ 、３５－８ ％ 及１２－５ ％ ,细胞增殖活性随肿瘤ｐ５３ 、ｂｃｌ２、ＭＤＭ２ 表达水平增高而增高,细胞凋亡则相反。结论：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白过表达与细胞增殖失控、凋亡抑制关系密切,在胶质瘤恶性进展中起重要作用。     

维生素A对胸腺凋亡有关基因的影响

为观察营养缺乏及补充维生素Ａ（ＶＡ）对胸腺细胞凋亡有关基因的影响，采用原位杂交及免疫组化的方法观察了胸腺组织ｂｃｌ－２ｍＲＮＡ，Ｆａｓ及Ｂａｘ蛋白的表达。结果表明营养缺乏的大鼠胸腺细胞ｂｃｌ－２ｍＲＮＡ的表达明显降低，而Ｆａｓ及Ｂａｘ蛋白的表达则明显增强；补充ＶＡ后大鼠胸腺ｂｃｌ－２ｍＲＮＡ的表达明显增强，Ｂａｘ蛋白的表达明显降低，而Ｆａｓ蛋白的表达无明显变化。本结果表明：营养缺乏时胸腺细胞凋亡增加导致免疫功能降低，补充ＶＡ可抑制细胞凋亡，可能是通过促进ｂｃｌ－２ｍＲＮＡ的表达，抑制Ｂａｘ蛋白的表达而起作用     

bcl—2基因在小鼠睾丸及试验隐睾中表达的研究

目的 研究ｂｃｌ－２基因在正常小鼠睾丈中的定位表达及试验隐睾所导致的改变。方法 以Ｗｅｓｔｅｒｎ－ｂｌｏｔｔｉｎｇ印迹法从蛋白水平检测ｂｃｌ－２基因在正常小鼠睾及及试验隐睾中的表达、变化;地高辛标记ｂｃｌ－２基因ｃＤＮＡ探针,石蜡组织切片原位杂交技术,从ｍＲＮＡ水平检测ｂｃｌ－２基因在小鼠精上皮中的定位及试验隐睾所导致的改变。结果 ｂｃｌ－２蛋白在正常小鼠睾丸中有较丰富的表达,试验隐睾导致其表达明显降低;ｂｃｌ－２基因在生精细胞中有较高水平的ｍＲＮＡ转录,表达细胞主要为各级生精细胞,支持细胞和间质细胞未见表达,隐睾术后仍然保持较高水平的ｍＲＮＡ转录。结论 ｂｃｌ－２蛋白可能在生精细胞主动退化的调节中起着重要作用。     

人肺癌细胞凋亡与p53、bcl-2、Bax的表达

目的：探讨人肺癌组织细胞自发凋亡的发生,ｐ５３基因及凋亡相关基因ｂｃｌ－２、Ｂａｘ在肺癌组织中的表达及与肺癌细胞凋亡和生物学行为的关系。方法：用甲基绿－派诺宁染色检测５０例肺癌组织切片中凋亡细胞,ＬＳＡＢ免疫组化法检测ｐ５３和Ｂａｘ的蛋白表达,原位杂交方法检测ｂｃｌ－２ ｍＲＮＡ的表达。结果：１００％小细胞肺癌凋亡发生数１０个／ＨＰＦ,２１％的肺鳞癌和１５％的肺腺癌凋亡发生数〉１０个／ＨＰＦ。ｐ５     

鼻咽癌组织中bcl—2,bax和p53的表达及其与瘤细胞？…

目的;了解鼻咽癌细胞中瘤细胞ｂｃｌ－２、ｂａｘ和ｐ５３的表达及其与瘤细胞凋亡指数的关系。方法：对３８例未经治疗的鼻咽癌组织,应用免疫组化ＬＳＡＢ法检测瘤ｂｃｌ－２、ｂａｘ和ｐ５３的表达;末端标记细胞死亡检测法（ＴＵＮＥＬ）计算癌细胞的凋亡指数。结果：（１）３７例（９７．４％）中有９０％左右的瘤细胞呈ｂｃｌ－２过表达;（２）３８例（１００％）中有７０％左右的瘤细胞过表达ｂａｘ;（３）２９例（７６．３     

乳腺癌和癌前病变中细胞凋亡及其与bcl-2、PCNA表达的关系

目的：探讨细胞凋乳腺癌变过程中的作用及其与细胞增殖以及ｂｃｌ－２、ＰＣＮＡ表达的关系。方法：利用ＴＵＮＥＬ法及免疫组化Ｓ－Ｐ法检测５４例乳腺癌及２７例非癌病变中细胞凋亡指数（ＡＩ）以及ｂｃｌ－２、ＰＣＮＡ的表达,同时计算核分裂指数（ＭＩ）。结果：“正常”乳腺上皮、增生性导管、原位主浸润性癌的Ａ少ＭＩ分别为：０．１０％±０．１２％、０．３１％±０．４３％、０．４１％±０．２１％、０．７４％±０．５６     

宫颈癌细胞增殖及凋亡与bcl-2表达的研究

本研究旨在探求宫颈癌变中细胞增殖和细胞凋亡的变化规律,以及凋亡出现频率与ｂｃｌ－２癌蛋白的关系。结果发现宫颈癌组织的增殖指数（ＰＩ）和凋亡指数（ＡＩ）明显高于正常宫颈组织和宫颈不典型增生组织（Ｐ＜０．０１）,而ＡＩ／ＰＩ显著低于两者（Ｐ＜０．０１）。ｂｃｌ－２癌蛋白常见于正常宫颈上皮和不典型增生上皮的基底细胞层,宫颈癌组织中其表达随临床期别升高呈下降趋势,ｂｃｌ－２阳性组的ＡＩ与阴性组无显著差异。证实宫颈癌变是细胞无限增殖,凋亡异常被抑制的结果。ｂｃｌ－２是宫颈癌变的早期特征,但不是调节细胞凋亡的唯一因素     

慢性缺氧对大鼠肺内细胞增殖、凋亡及相关基因表达的影响

目的：探讨慢性缺氧性肺动脉高压肺血管结构改建（ｐｕｌｍｏｎａｒｙｖａｓｃｕｌａｒｓｔｒｕｃｔｕｒａｌｒｅｍｏｄｅｌｉｎｇ，ＰＶＳＲ）的细胞及分子机制。方法：大鼠慢性缺氧模型复制，免疫组化及Ｎｏｒｔｈｅｒｎ杂交检测大鼠肺内ｃ－ｍｙｃ及ｂｃｌ－２两基因蛋白及ｍＲＮＡ的表达，免疫组化及原位缺口末端标记法检测大鼠肺内增殖相关抗原Ｋｉ－６７及凋亡细胞。结果：正常及慢性缺氧大鼠肺内均检出一定比率的增殖或凋亡细胞，缺氧１、２周大鼠肺内细胞增殖／凋亡比值比对照组分别增高约３、３５倍。与细胞增殖、凋亡密切相关的ｃ－ｍｙｃ、ｂｃｌ－２基因表达在缺氧１周大鼠肺内显著增高，在缺氧２周更显著。结论：细胞增殖、凋亡机制共同参与了慢性缺氧性肺动脉高压时ＰＶＳＲ的调节。     

反义bcl—2寡脱氧核苷酸对U937细胞增殖和存活能力的影响

本实验观察了与人类ｂｃｌ－２ ｍＲＮＡ翻译起始部位相互补的硫代反义ｂｃｌ－２寡脱氧核苷酸（ＡＳ－ｏＯＤＮ）对Ｕ９３７细胞增殖和存活能力的影响，结果表明：一定浓度的ＡＳ－ｓＯＤＮ可明显抑制细胞Ｂｃｌ－２蛋白的表达，但对细胞增殖和存活能力无明显影响。这一研究结果为在不同表达ｂｃｌ－２的肿瘤细胞中探讨ｂｃｌ－２反义核酸作用的普遍性和正确评价其作用地位积累了资料。     

淋巴瘤凋亡调节基因bcl—x mRNA及蛋白的表达

目的 探讨凋亡调节基因ｂｃｌ－ｘ在淋巴瘤细胞凋亡调控和肿瘤发生中的作用及在淋巴瘤诊断中的价值。方法 收集３０例新鲜标本和１０９例存蜡块（均包括淋巴反应性增生和Ｔ、Ｂ常见类型淋巴瘤）,用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）和免疫组化枸橼酸－微波－ＡＢＣ法分别观察ｂｃｌ－ｘ ｍＲＮＡ和其蛋白的表达及分布。结果 ＲＴ－ＰＣＲ显示３０例新鲜标本中均有ｂｃｌ－ｘ的转录,其中２９例可见ｂｃｌ－ｘ条带,１３例出     

胶质瘤细胞ING1、人端粒酶逆转录酶及人端粒酶相关蛋白1基因表达的研究

目的 探讨胶质瘤细胞ING1、人端粒酶逆转录酶(hTERT)和人端粒酶相关蛋白1(hTP1)基因表达的意义。方法 用mRNA原位杂交及免疫组织化学染色方法观察了70例不同级别的人胶质瘤组织。结果 hTERT mRNA和蛋白的阳性表达率分别为88.6％和82.9％,hTP1 mRNA和蛋白的阳性表达率均为100％,这4种阳性肿瘤细胞的密度彼此间均呈正相关(r=0.758～0.882,P<0.0005),并均随肿瘤级别升高而相应增加(P<0.05～0.01)。ING1 mRNA和蛋白的阳性表达率分别为94.3％和88 6％,两种阳性肿瘤细胞密度间也呈正相关(r=0.831,P<0.0005),但两者均随肿瘤级别升高而相应减少(P<0.01),并均分别与hTERT mRNA和蛋白及hTP1 mRNA和蛋白的阳性肿瘤细胞密度呈负相关(r=-0.211～-0.384,P<0.05～0.001)。结论 胶质瘤细胞中hTERT和hTP1基因表达异常增加可能是抑制其ING1基因表达的重要因素,这3种基因的表达异常可能在胶质瘤发生及恶性进展过程中起重要作用。     

caspase-3、bcl-2蛋白在非霍奇金淋巴瘤组织的表达及其与细胞凋亡和增殖的关系

目的探讨caspase-3、bcl-2蛋白在非霍奇金淋巴瘤(NHL)发生、发展中的可能作用及相互关系.方法应用TdT介导的dUTP缺口末端标记(TUNEL)技术和免疫组织化学链霉素抗生物素-过氧化酶(SP)法,检测5例反应性增生淋巴组织和119例NHL组织中的细胞凋亡和增殖细胞核抗原(PCNA)、caspase-3、bcl-2蛋白的表达水平.结果 caspase-3和bcl-2在119例NHL中表达的阳性率分别为86.6%(103例)和53.8%(64例).二者在良、恶性淋巴组织中的表达方式相反在反应性增生淋巴滤泡中心caspase-3高表达而bcl-2阴性表达,滤泡套区caspase-3阴性表达而bcl-2高表达;在肿瘤性滤泡中心bcl-2常高表达而caspase-3常表达减弱或不表达;在NHL中二者的表达与肿瘤恶性度呈相反关系,高度恶性组中caspase-3的表达(44.4%)高于低度恶性组(23.7%,P<0.01),而B细胞性淋巴瘤中bcl-2的表达(42.6%)低于低度恶性组(75.5%,P<0.01);caspase-3的表达与凋亡指数呈正相关(r=0.512, P <0.01)),bcl-2的表达与凋亡指数呈负相关(r=-0.436, P<0.01).此外,NHL的凋亡指数与增殖指数呈显著正相关(r=0.710, P<0.01).结论 caspase-3可能参与了NHL的凋亡调节机制.caspase-3和bcl-2蛋白在良、恶性淋巴组织中常呈现相反的表达方式,提示二者在淋巴细胞增殖动力学调节中可能存在着密切联系.     

Upregulation of SATB1 is associated with the development and progression of glioma

ABSTRACT: BACKGROUND: Special AT-rich sequence-binding protein-1 (SATB1) has been reported to be expressed in several human cancers and may have malignant potential. This study was aimed at investigating the expression and potential role of SATB1 in human glioma. METHOD: The relationship between SATB1 expression, clinicopathological parameters, Ki67 expression and MGMT promoter methylation status was evaluated, and the prognostic value of SATB1 expression in patients with gliomas was analyzed. SATB1-specific shRNA sequences were synthesized, and U251 cells were transfected with SATB1 RNAi plasmids. Expression of SATB1 mRNA and protein was investigated by RT-PCR and immunofluoresence staining and western blotting. The expression of c-Met, SLC22A18, caspase-3 and bcl-2 protein was determined by western blotting. U251 cell growth and adherence was detected by methyl thiazole tetrazolium assay. The apoptosis of U251 cells was examined with a flow cytometer. The adherence, invasion, and in vitro angiogenesis assays of U251 cells were done. The growth and angiogenesis of SATB1 low expressing U251 cells was measured in an in vivo xenograft model. RESULTS: Of 70 tumors, 44 (62.9%) were positive for SATB1 expression. SATB1 expression was significantly associated with a high histological grade and with poor survival in univariate and multivariate analyses. SATB1 expression was also positively correlated with Ki67 expression but negatively with MGMT promoter methylation in glioma tissues. SATB1 shRNA expression vectors could efficiently induce the expression of SLC22A18 protein, increase the caspase-3 protein, inhibit the expression of SATB1, c-Met and bcl-2 protein, the growth, invasion, metastasis and angiogenesis of U251 cells, and induce apoptosis in vitro. Furthermore, the tumor growth of U251 cells expressing SATB1 shRNA were inhibited in vivo, and immunohistochemical analyses of tumor sections revealed a decreased vessel density in the animals where shRNA against SATB1 were expressed. CONCLUSION: SATB1 may have an important role as a positive regulator of glioma development and progression, and that SATB1 might be a useful molecular marker for predicting the prognosis of glioma.     

Clinicopathological significance of Polo-like kinase 1 (PLK1) expression in human malignant glioma

Polo-like kinase 1 (PLK1), a variety of serine/threonine-protein kinase, has been reported to play important roles in malignant transformation. The purpose of this study was to investigate the clinicopathological significance of PLK1 expression in malignant glioma. A semi-quantitative RT-PCR assay was performed to detect the expression of PLK1 mRNA in 68 cases of glioma tissues and corresponding non-cancerous brain tissues. Additionally, the correlation of PLK1 mRNA expression with clinicopathological factors or prognosis of glioma patients was statistically analyzed. Multivariate analysis of prognostic factors was performed using the Cox proportional hazard model. Small interfering RNA was used to knockdown PLK1 expression in a glioma cell line and analyze the effects of PLK1 inhibition on growth, cell cycle, apoptosis and chemo- or radiosensitivity of glioma cells. Results showed that the expression of PLK1 mRNA was significantly higher in glioma tissues than in corresponding normal brain tissues. The expression of PLK1 mRNA was closely correlated with WHO grade, KPS and tumor recurrence of glioma patients (P = 0.022, 0.030 and 0.041, respectively). Meanwhile, the disease-free and overall survival rates of patients with high PLK1 mRNA expression were obviously lower than those of patients with low PLK1 mRNA expression. Multivariate analysis showed that high PLK1 mRNA expression was a poor prognostic factor for glioma patients (P = 0.028). The expression of PLK1 mRNA and protein was significantly down-regulated in stably transfected U251-S cells. PLK1 down-regulation could inhibit growth, induce cell arrest in G2/M phase of cell cycle and apoptosis enhancement in glioma cells. Further, PLK1 down-regulation could enhance the sensitivity of glioma cells to cisplatin or irradiation. Thus, the status of PLK1 mRNA expression might be an independent prognostic factor for glioma patients and targeting PLK1 could be a novel strategy for chemo- or radiosensitization of human malignant gliomas.     

CD95 Ligand: Lethal Weapon Against Malignant Glioma?

CD95 (Fas/APO-1) and its ligand (CD95L) belong to a growing cytokine and cytokine receptor family that includes nerve growth factor (NGF) and tumor necrosis factor (TNF) and their corresponding receptors. CD95 expression increases during malignant progression from low-grade to anaplastic astrocytoma and is most prominent in perinecrotic areas of glioblastoma. There is, however, no evidence that CD95 expression in malignant gliomas is triggered by hypoxia or ischemia. Agonistic antibodies to CD95, or the natural ligand, CD95L, induce apoptosis in human malignant glioma cells in vitro. Glioma cell sensitivity to CD95-mediated apoptosis is regulated by CD95 expression at the cell surface and by the levels of intracellular apoptosis-regulatory proteins, including bcl-2 family members. Several cytotoxic drugs synergize with CD95L to kill glioma cells. For as yet unknown reasons, glioma cells may co-express CD95 and CD95L in vitro without undergoing suicide or fratricide. Yet, they kill T cells via CD95/CD95L interactions and are sensitive to exogenously added CD95L. Since CD95L is expressed in gliomas in vivo , too, forced induction of CD95 expression might promote therapeutic apoptosis in these tumors. That glioma cells differ from non-transformed T cells in their sensitivity to CD95 antibodies or recombinant ligand, may allow the development of selective CD95 agonists with high antitumor activity that spare normal brain tissue. A family of death ligand/receptor pairs related to CD95L/CD95, including APO2L (TRAIL) and its multiple receptors is beginning to emerge. Although several issues regarding glioma cell sensitivity to CD95L/CD95-mediated apoptosis await elucidation, CD95 is a promising target for the treatment of malignant glioma.     

Bcl-2、Bax在NHE-1抑制导致的大鼠肺动脉平滑肌细胞凋亡中的作用

目的:探讨Bcl-2、Bax蛋白表达在Na⁺/H⁺交换器-1(NHE-1)抑制而诱导大鼠肺动脉平滑肌细胞(PASMC)凋亡中的作用。方法: 荧光指示剂(Fura-2/AM)测定法检测转染NHE-1特异性核酶基因的大鼠PASMC内Ca²⁺(i)变化;RT-PCR方法检测细胞内bcl-2和baxmRNA表达变化, 免疫组化法检测细胞内Bcl-2和Bax蛋白表达变化。 结果:转染NHE-1特异性核酶基因后, 大鼠PASMC内i显著升高, bcl-2mRNA及蛋白表达显著降低, baxmRNA和蛋白表达显著增加。结论: NHE-1抑制诱导的PASMC凋亡与i增加、bcl-2表达降低及bax表达增加有关。     

反义bcl-2硫代磷酸寡脱氧核苷酸对小细胞肺癌细胞株NCI-H446的抑制作用及诱导细胞凋亡的研究

目的:观察反义bcl-2硫代磷酸寡脱氧核苷酸(AS-PS-ODN)对小细胞肺癌细胞株NCI-H446mRNA、蛋白以及增殖、活力和凋亡的影响。方法: 合成bcl-2AS-PS-ODN作用于小细胞肺癌细胞株NCI-H446, 半定量RT-PCR检测bcl-2mRNA表达;免疫细胞化学染色和流式细胞仪检测Bcl-2蛋白表达;通过克隆形成率、细胞计数、流式细胞仪DNA倍体分析、TUNEL等指标观察bcl-2 AS-PS-ODN对细胞增殖、活力和凋亡的影响。结果:①bcl-2 AS-PS-ODN能特异性地降低NCI-H446细胞bcl-2mRNA和Bcl-2蛋白的表达。1μmol/LAS-PS-ODN作用24h后bcl-2mRNA表达量下降69.5%, 48h后Bcl-2蛋白下降62.7%。②bcl-2 AS-PS-ODN能够抑制细胞增殖和活力, 诱导细胞凋亡, 1μmol/LAS-PS-ODN作用24h后, 细胞凋亡率约为22.3%-32.7%。结论:bcl-2 AS-PS-ODN能够特异性降低bcl-2mRNA、蛋白表达, 抑制NCI-H446细胞的增殖和活力, 并诱导细胞凋亡。     

Molecular mechanism of SSFA2 deletion inhibiting cell proliferation and promoting cell apoptosis in glioma

Gliomas are the most common primary brain malignant tumors in humans. Glioblastoma multiforme(GBM) is the most malignant intracranial tumor with a relatively poor prognosis. There promote us to find effective anti-cancer therapies to reduce cancer mortality. By using bioinformatic analysis, we found SSFA2 as a gene with elevated expression in the glioma tissues. We detected the expression of SSFA2 in glioma tissues and in the glioma cell lines, as well as in normal brain tissues. SSFA2 expression was higher in glioma tissues, especially in glioblastoma multiforme than normal brain tissues. Subsequently, we found that down-regulate SSFA2 in glioma cell lines can regulate the cell cycle to reduce the proliferation ability and induce the early apoptosis rate in shSSFA2 cells relative to control cells. Moreover, we found that down-regulate SSFA2 in glioma cell line U87(shSSFA2-U87) inhibited the growth effectiveness compared to the control cell line U87. These result reveals us that SSFA2 may act as oncogene to promote the progression of glioma. For further research specific mechanisms of SSFA2 in gliomas, we used the gene chip to detect the downstream gene in U87. We found that 30 genes also may be as target gene of SSFA2, and we testify the protein expression by western-blot. The result reveal that IL1A, IL1B and CDK6 as target gene of SSFA2 to regulate the progression of glioma. These finding suggest that SSFA2 could be a new therapeutic target for gliomas.     

Participation of host cells: resistance or collaboration

Matrix metalloproteinases play an important regulatory role in tissue morphogenesis, cell differentiation and motility, and tumor cell invasiveness. We have recently demonstrated elevated activity of the 92 kDa type IV collagenase (MMP-9) in human glioblastoma and in the present study examine the relative amounts of MMP-9 protein and mRNA in human gliomas and as well as the distribution of MMP-9 in human glioma tumors in vivo. Using an enzyme-linked immunosorbent assay for the quantitative determination of MMP-9 protein, we found that levels were significantly higher in malignant astrocytomas, especially in glioblastoma multiforme, than in normal brain tissues and low-grade gliomas. In addition, the amount of MMP-9 mRNA, as determined by northern blot analysis was higher in anaplastic astrocytomas and glioblastoma multiforme than in normal brain tissue and low-grade gliomas. Immunocytochemical staining for MMP-9 showed strong cytoplasmic immunoreactivity in the tumor cells and the proliferating endothelial cells of glioblastoma multiforme and anaplastic astrocytomas. The staining intensity was lower in low-grade astrocytomas, and was undetectable or very low in normal brain astrocytes. The results indicate that expression of MMP-9 is dramatically upregulated in highly malignant gliomas and correlates with the highly malignant progression of human gliomas in vivo, and support a role for the MMP-9 in facilitating the invasiveness seen in malignant gliomas in vivo.