17-β雌二醇对宫颈癌Hela细胞环氧合酶-2和增殖细胞核抗原表达的影响

目的:探讨17-β雌二醇(E2)对宫颈癌Hela细胞环氧合酶-2(cyclooxygenase-2,COX-2)和增殖细胞核抗原(proliferatingcell nuclear antigen,PCNA)表达的影响。方法:Hela细胞分别用0.1%乙醇(对照组)、E2(E2组)、E2+依托昔布(E2+依托昔布组)作用24 h后,采用免疫细胞化学、Western blot及RT-PCR方法检测COX-2和PCNA的表达。结果:E2组COX-2和PCNA蛋白及mRNA水平较对照组明显升高(P0.01);而E2+依托昔布组PCNA水平较E2组显著降低(P0.01),但依然高于对照组。结论:E2可促进Hela细胞COX-2表达,并进一步上调PCNA水平。     

内皮素-1对前列腺癌PC3细胞环氧化酶-2表达的影响

目的:探讨在激素非依赖性前列腺癌(HRPC)PC3细胞中,内皮素-1(ET-1)刺激对环氧化酶-2(COX-2)表达的影响,以及内皮素受体A(ETAR)与内皮素受体B(ETBR)在此调控通路中的作用.方法:用不同浓度(0.1、1.0、10.0、100.0 nmol/L)ET-1刺激前列腺癌PC3细胞相同时间(24 h);另外用BQ123(ETAR拮抗剂)、BQ788(ETBR拮抗剂)单独作用于PC3细胞或与100 nmoL/L ET-1联合作用于细胞24 h.均采用Western blot技术检测细胞中COX-2蛋白含量的变化:采用RT-PCR技术检测细胞中COX-2 mRNA的变化情况.结果:PC3细胞在0.1-100.0 nmol/L浓度的ET-1刺激后COX-2的蛋白及mRNA表达水平均显著上调,并显示有明显的浓度依赖性.BQ123在蛋白质和mRNA水平均显著抑制ET-1介导的COX-2的上调,与单纯应用ET-1组有显著差别(P<0.05),BQ788则对ET-1介导的COX-2的表达没有抑制作用(P>0.05).结论:在前列腺癌PC3细胞中,ET-1对COX-2的表达有上调作用;ETAR参与厂这一调控过程,ETBR可能未参与此过程.这为ETAR拮抗剂和选择性COX-2抑制剂联合应用于HRPC的治疗提供了分子生物学基础.     

PTEN影响宫颈癌Hela细胞对顺铂的化疗敏感性

目的:通过转染PTEN基因表达载体,提高宫颈癌Hela细胞中PTEN的表达水平,观察能否影响Hela细胞对顺铂的敏感性,并探讨其分子学机制.方法:将培养的宫颈癌Hela细胞分为空白对照(未转染质粒的Hela细胞)、质粒对照(转染空质粒的Hela细胞)和PTEN转染(转染PTEN表达载体的Hela细胞)3组.分别用RT-PCR和Western检测各组Hela细胞中PTEN的mRNA和蛋白的表达水平;MTT法检测各组Hela细胞对顺铂的化学敏感性;RT-PCR检测各组Hela细胞中磷脂酰肌醇3-激酶(PI3K)的表达水平.结果:PTEN转染组Hela细胞中PTEN的表达水平比另2组细胞显著增高(P＜0.01);经顺铂处理后,PTEN转染组Hela细胞生长抑制率比其它2组显著增高(P＜0.01),PTEN转染组Hela细胞中PI3K被显著抑制(P＜0.01).结论:PTEN能增强宫颈癌Hela细胞对顺铂的化疗敏感性,它可能通过下调Hela细胞中PI3K而导致Hela细胞对顺铂耐药性下降.     

表皮生长因子对PC-3细胞环氧化酶-2表达的影响

目的:探讨表皮生长因子(EGF)对人激素非依赖性前列腺癌PC-3细胞环氧化酶-2(COX-2)表达的影响及可能的机制.方法:以1、10、100 μg/L不同浓度的EGF作用于PC-3细胞24 h,采用逆转录聚合酶链反应(RT-PCR)和Western blot测定COX-2的表达变化;再将磷脂酰肌醇-3激酶阻断剂(LY294002,20μmol/L)、促分裂原活化蛋白激酶阻断剂(SC203580,20 μmol/L)和表皮生长因子受体阻断剂(ZD1839,2 μmol/L)分别与10μg/L EGF作用于PC-3细胞,Western blot测定COX-2蛋白的变化.结果:PC-3细胞在1～100 μg/L浓度的EGF刺激以后的表达显著上调,和对照组有明显区别(P＜0.05).SC203580、LY294002和ZD1839显著抑制COX-2上调(P＜0.05).结论:EGF可诱导PC-3细胞COX-2的表达上调,促分裂原活化蛋白激酶和磷脂酰肌醇-3激酶参与了其中的调节过程.     

JMJD2B、HPIP对宫颈癌Hela细胞生物学行为的影响

目的分析JMJD2B、HPIP对宫颈癌Hela细胞生物学行为的影响。 方法将Hela细胞分为si-NC组、si-JMJD2B组和si-HPIP组。实时荧光定量PCR检测宫颈癌细胞中JMJD2B、HPIP mRNA表达水平。Western blotting检测宫颈癌细胞和人正常宫颈上皮细胞JMJD2B、HPIP蛋白表达水平。CCK-8检测Hela细胞增殖能力。Transwell检测Hela细胞迁移和侵袭能力。 结果JMJD2B、HPIP蛋白在宫颈癌细胞中表达高于人正常宫颈上皮细胞(P＜0.05)。与si-NC组比较,si-JMJD2B组和si-HPIP组JMJD2B、HPIP mRNA和蛋白表达、细胞增殖、迁移细胞数量、侵袭细胞数量均明显减少(P＜0.05)。 结论JMJD2B和HPIP表达可能调节宫颈癌Hela细胞增殖、迁移和侵袭等生物学行为。     

胃癌不同临床病理分期中环氧合酶-2与基质金属蛋白酶-2的表达及其相关性

目的分析环氧合酶-2（COX-2）与基质金属蛋白酶-2（MMP-2）在胃癌不同临床病理分期中的表达及其相关性。方法采用免疫组化方法检测90例胃癌原发灶标本中COX-2及MMP-2的表达情况,观察其与胃癌临床病理分期的关系,并分析COX-2与MMP-2在胃癌组织中表达的相关性。结果90例胃癌标本中,COX-2和MMP-2的表达阳性率分别为74．4％（62／90）和68．9％（62／90）;COX-2和MMP-2的表达与胃癌TNM分期、局部淋巴结转移及远处转移密切相关（P〈0．05）,COX-2还与肿瘤的分化程度相关;此外,COX-2的表达与MMP-2的表达存在明显相关关系（r=0．749,P〈0．01）。结论胃癌组织中COX-2和MMP-2的表达与胃癌临床病理分期密切相关;胃癌组织中COX-2的表达与MMP-2的表达有明显相关。     

肺癌组织中环氧化酶-2表达和临床病理特征关系的研究

目的:研究肺癌组织中环氧化酶-2(COX-2)的表达及其与临床病理特征、预后的关系.方法:采用免疫组化SABC法检测58例肺癌组织及9例癌远旁正常肺组织中COX-2表达.结果:肺癌组织COX-2阳性表达为60.34%(35/58)较正常肺组织11.11%(1/9)差异有统计学意义(P＜0.01);COX-2的表达与患者的TNM分期、淋巴结浸润显著相关(P＜0.05),与患者的年龄、性别、吸烟状况、组织类型无关(P＞0.05).COX-2阳性表达组的生存率低于阴性表达组(P=0.001).结论:COX-2在肺癌的发展、转移过程中有一定作用,COX-2可以作为肺癌的预后指标.     

Expression of Cyclooxygenase-2 in Ovarian Cancer Cell Lines

Summary： To investigate the expression of cyclooxygenase-2 （COX-2） in ovarian cancer cell lines, RT-PCR and immunocytochemistry were used to detect the expression of COX-2 in 5 ovarian cancer cell lines. The expression of COX-2 mRNA and protein was detected in all 5 cell lines. It is suggested that COX-2 is expressed in ovarian cancer cell lines, which provides a Basis for the chemoprevention of ovarian cancer.     

精胺氧化酶高表达对人宫颈癌Hela细胞生长和药物敏感性的影响

目的 研究精胺氧化酶(SMO)稳定性高表达对人宫颈癌Hela细胞生长和药物敏感性的影响.方法 含SMO全长编码序列的真核表达质粒用脂质体法稳定转染人宫颈癌Hela细胞后,QT-RT-PCR和酶活性分析法筛选SMO高表达的细胞株.MTS法检测细胞的增殖和对抗癌药物的敏感性.流式细胞分析法检测细胞周期和细胞凋亡.结果 成功获得SMO稳定高表达的人Hela宫颈癌细胞株.MTS分析发现,SMO高表达细胞的生长速度显著性高于对照细胞,且对多胺类似物抗癌药物CPENSpm的敏感性明显下降.与对照细胞相比,CPENSpm诱导SMO高表达细胞发生细胞凋亡的能力明显减弱.结论 SMO稳定性高表达可降低宫颈癌细胞对抗癌药物CPENSpm的敏感性.     

家族性腺瘤性息肉病中COX-2的表达分析

目的:分析环氧化酶2(COX-2)在家族性腺瘤性息肉病(FAP)中的表达,探讨COX-2在腺瘤形成以及癌变过程中的可能作用.方法:收集我科2001年1月至2003年6月10例FAP患者手术切除标本,分别取≤0.5 cm、≥1 cm的腺瘤性息肉以及癌变腺瘤组织,采用EnVision免疫组化方法观察COX-2在这些病灶中的表达.结果:COX-2主要在上皮细胞内表达,在部分间质细胞中也表达.COX-2在≤0.5 cm腺瘤上皮细胞中的表达强于正常上皮,但相差并不显著;在≥1.0 cm腺瘤中表达显著强于正常上皮(P＜0.01);在不同大小腺瘤之间的表达差异显著,≥1.0 cm腺瘤中表达强于≤0.5 cm腺瘤(P＜0.01);癌变腺瘤中的COX-2表达显著高于正常上皮(P＜0.01);癌变腺瘤中表达强于≥1.0cm的腺瘤组织,但相差并不显著.结论:COX-2可能是FAP发展中的重要促进因子,参与了腺瘤早期形成以及癌变过程;选择性抑制COX-2是预防治疗FAP的有效靶点.     

体外培养的皮层神经元和胶质细胞上COX-2的表达

目的观察环氧合酶-2（cyclooxygenase-2,COX-2）在体外培养的皮层神经元和胶质细胞上的表达。方法在体外培养鼠的皮层神经元和神经胶质细胞,神经元培养第6、10、12d时收获细胞,神经胶质细胞传代后培养待细胞密度约为每毫升5×105时和6×105时收获细胞,然后进行免疫染色以观察神经元和胶质细胞上COX-2的表达。同时用免疫荧光双标记法对COX-2阳性的细胞类型进行了鉴定。结果体外培养6、10和12d时的胚鼠皮层神经元细胞均呈现OX-2免疫阳性,但不同时间点免疫染色强度不同,10d时免疫染色最强;而体外培养的新生鼠的皮层神经胶质细胞只有极个别细胞呈现COX-2免疫阳性;双标记结果表明这些胶质细胞既有星形胶质细胞,又有小胶质细胞。结论离体条件下COX-2主要在神经元上表达,而神经胶质细胞则很少表达,与在体研究结果基本一致。     

光动力联合塞来昔布对子宫颈癌COX-2表达及影响的研究

目的研究光动力联合塞来昔布对子宫颈癌细胞中环氧合酶-2(cyclooxynase-2,COX-2)蛋白表达及细胞增殖和凋亡的影响。方法①培养子宫颈癌Hela细胞,应用四甲基偶氮唑蓝(MTT)比色法检测选择性COX-2抑制剂塞来昔布(celecoxib)对子宫颈癌Hela细胞增殖抑制作用。②将Hela细胞分为4组:空白对照组(Hela,H组)、塞来昔布组(celecoxib,C组)、光动力组(photodynamics therapy,PDT,P组)、塞来昔布+光动力组(celecoxib+photodynamics therapy,P+C组)、分别应用蛋白印迹法(Westernblot)和免疫组化法检测COX-2蛋白表达情况。结果 MTT比色法检测显示,celecoxib作用Hela细胞24 h后对宫颈癌细胞有相对的抑制作用。Western blot和免疫组化法检测结果均显示:celecoxib联合PDT作用Hela细胞24 h后COX-2蛋白的相对表达水平明显降低,与其它3组相比,差异有统计学意义(P<0.05)。结论选择性COX-2抑制剂celecoxib能抑制人子宫颈癌细胞的增殖,且呈浓度依赖性,随着浓度的增大作用逐渐增强。PDT联合选择性COX-2抑制剂celecobix能有效抑制靶基因COX-2的表达,进而可抑制子宫颈癌细胞增殖并诱导细胞调亡增加,为子宫颈癌的基因研究及治疗提供新思路。     

环氧合酶2启动子区-899G/C 基因多态性与中国乙肝相关性肝癌的关系

背景：肝细胞肝癌的发病与乙型肝炎病毒感染密切相关,研究发现环氧合酶2（COX-2）在肝癌中高表达,并参与了肝癌的发生。本研究通过分析COX-2启动子区-899G/C基因多态性在中国甘肃省健康人群、乙肝病毒性肝炎、乙后肝硬化、原发性肝癌患者的分布,旨在探讨COX-2 -899G/C基因多态性与中国甘肃省乙肝相关易感性肝癌的关系。 方法：采用PCR-TaqMan 探针方法检测甘肃省健康人群、乙肝病毒性肝炎、乙肝后肝硬化、原发性肝癌患者COX-2启动子区-899G/C的基因多态性。 结果：COX-2-899G/C分为GG、GC、CC三种基因型,其频率在乙肝病毒性肝炎患者中分别为：87.00%, 12.67%, 0.33%;在乙后肝硬化患者中分别为：85.33%, 14.00%, 0.67%;在原发性肝癌患者中分别为：77.00%, 21.67%, 1.33%;在普通人群中分别为: 8790.67%, 9.00%, 0.33%。与GG基因型相比,COX-2-899 C等位基因携带者患乙肝相关易感性性肝癌的风险增加。 结论：COX-2-899G/C的C等位基因增加我国肝癌高发地区甘肃省乙肝相关易感性肝癌发病的风险。     

苦参碱通过P38通路调节H2AX磷酸化抑制宫颈癌细胞的增殖及迁移

目的 探讨苦参碱对宫颈癌细胞增殖、凋亡、迁移的影响及其可能的分子机制。方法 不同浓度苦参碱作用于宫颈癌细胞,通过CCK8法,EdU法、流式细胞术、Transwell实验、划痕实验检测细胞的增殖、凋亡、迁移能力。利用Western印迹法检测苦参碱对H2AX磷酸化蛋白（即γH2AX）、P38磷酸化蛋白（即pP38）水平的影响。通过干扰H2AX（Ser139）位点的磷酸化,检测苦参碱对宫颈癌细胞增殖、迁移的影响。通过干扰P38磷酸化,检测苦参碱对H2AX磷酸化的影响。结果 苦参碱对宫颈癌细胞体外增殖具有显著抑制作用（P<0.05）,且呈药物浓度依赖性;苦参碱显著提高细胞凋亡率,而对迁移能力具有显著抑制作用（P<0.05）;苦参碱能够显著促进γH2AX及pP38蛋白的表达（P<0.05）,且呈浓度依赖性。H2AX（Ser139）位点的磷酸化是苦参碱发挥抑增殖和迁移的作用位点。苦参碱通过P38通路调控H2AX（Ser139）位点的磷酸化。结论 苦参碱抑制宫颈癌细胞的增殖和迁移,诱导细胞凋亡,可能是通过调控P38通路使H2AX（Ser139）位点磷酸化发挥对宫颈癌的抑癌作用。     

The role of cyclooxygenase-2/prostanoid pathway in visceral

Background Cyclooxygenase (COX) is the rate-limiting enzyme in the production of prostanoids from arachidonic acid. COX-2 is the inducible enzyme in the COX family, together with the prostanoids forms the COX-2/prostanoid pathway. Research showed that the COX-2/prostanoid pathway is activated in hepatic diseases and liver stress reaction, such as fibrogenesis, portal hypertension, carcinogenesis, and ischemic/reperfusion injury. But there was no report on visceral pain induced liver stress. This study was to investigate the role of the COX-2/prostanoid pathway in liver stress response in rat acute colitis visceral pain liver stress model. Methods Fifty-three male SD rats were randomly divided into Naive, Model, NS398 treatment, and Morphine treatment groups. The rat acute colitis visceral pain liver stress model was established under anesthesia by the colonic administration of 0.5 ml of 6% acetic acid using a urethral catheter. NS398 and morphine were administrated 30 minutes prior to model establishment in NS398 and Morphine treatment groups respectively. Spontaneous activities and pain behavior were counted and the extent of colonic inflammation was assessed histologically. Liver tissue levels of Glutathione-S-Transferase (GST) activity, COX-2 mRNA, prostaglandin E2 (PGE2), thromboxane B2 (TXB2) and 6-Ketone-prostaglandin F1α (6-K-PGF1α) contents were assessed.Results Thirty minutes after the colonic administration of acetic acid, a significant decrease in spontaneous activities and an increase in pain behaviors were observed in Model group (P&lt;0.01 and P&lt;0.05 respectively), accompanied by colonic inflammation. Liver GST activity levels significantly dropped (P&lt;0.05). Liver COX-2 mRNA expression significantly increased, accompanied by an increase in liver concentrations of PGE2 and TXB2, but no obvious change in 6-K-PGF1α concentrations. NS398 and morphine both ameliorated post-stress liver GST activity (P&lt;0.05 and P&lt;0.01 respectively), decreased stress-induced COX-2 expression, decreased PGE2 and TXB2 production, but increased liver 6-K-PGF1α levels. Morphine attenuation in colonic tissue inflammation was apparent at 24 hours (P&lt;0.05).Conclusions Acute colitis visceral pain liver stress can induce liver injury. Liver injury might have occurred through the activation of the COX-2/prostanoid pathway and increased production of PGE2 and TXB2. Effective analgesia might offer protective effect during visceral pain stress.     

EGFR和COX-2在PBM患者胆囊黏膜上皮中的表达及意义

目的探讨表皮生长因子受体（epidermal growth factor receptor,EGFR）和环氧合酶-2（cyclooxygenase-2,COX-2）在胰胆管汇合异常（pancreaticobiliary maljunction,PBM）患者胆囊黏膜上皮中表达及临床意义。方法应用免疫组化S-P法检测PBM和非PBM慢性胆囊炎患者胆囊黏膜上皮中COX-2和EGFR蛋白的表达。结果 22例PBM患者胆囊黏膜上皮中,COX-2表达为（-）2例,（＋）3例、（＋＋）6、（＋＋＋）11例,对照组（-）4例、（＋）9例、（＋＋）8例、（＋＋＋）1例;PBM患者胆囊黏膜上皮中EGFR表达为（-）2例,（＋）2例、（＋＋）9例、（＋＋＋）9例,对照组为（-）6例、（＋）4例、（＋＋）10例、（＋＋＋）2例。两者均较对照组表达增高,差异有统计学意义（P〈0.05）,且PBM患者胆囊黏膜上皮中COX-2和EGFR表达有相关性（r=0.584,P〈0.05）。结论 COX-2和EGFR在胰胆管汇合异常患者胆囊黏膜上皮中表达升高,且有相关性,可能对PMB患者胆囊上皮过度增生-不典型增生-癌变过程有促进作用。     

Induction of cyclooxygenase-2 by insulin in cultured rat vascular smoothmuscle cells

To evaluate cyclooxygenase-2 (COX-2) expression induced by insulin in cultured rat vascular muscle smooth cells (VSMCs). Methods: The rat VSMCs were isolated and cultured in 60 nma plates, pre-treated with insulin 10, 100, 500, 1 000 mU/L for 6 h, and 100 mU/L for 3, 6. 12, and 24 h, respectively. RT-PCR was performed to measure COX-2 mRNA induction, and Western-blot was used to measure protein expression. In regard to the COX-2 enzyme activity, after stimulation for a specified duration, the culture medium was collected ; Prostaglandin E2( PCE2 ) released by VSMCs was measured with an EIA kit. Results: Three hours after insulin ( 100mU/L) stimulation, the induction of COX-2 mRNA was detectable and increased with the time and dosage of insulin. Insulin induced COX-2 protein expression occurred in a time- and dose-dependent manner. In rat VSMCs, COX-2 induced by insulin resulted in PGE2 production, which was increased with insulin action. Conclusion: Insulin could induce COX-2 expression and PGF2, production in VSMCs in time and dose-dependent manners.     

Effects of aspirin on atherosclerosis and the cyclooxygenase-2 expression in atherosclerotic rabbits

Background Atherosclerosis is a complex vascular inflammatory disease. Aspirin is a mainstay in the prevention of vascular complications of atherosclerosis. In this study, the effectiveness of aspirin in suppressing atherosclerosis and the inflammation process was evaluated in rabbits fed with a high fat diet.Methods　Eighteen male New Zealand rabbits were randomly divided into 3 groups: control group, untreated cholesterol-fed group, aspirin treated cholesterol-fed group, which were fed for 12 weeks. After 12 weeks, the aorta was harvested for pathologic morphology observation. Immunohistochemical analysis of cyclooxygenase-2 (COX-2), macrophage and vascular smooth muscle cell (VSMC) was performed. The statistical analysis was performed by the statistical program SPSS10.0.Results　The aorta plaque/intima size (P/I) by pathologic morphology observation was 0%, (59.6±13.7)% and (36.3±16.5)% in the control, untreated cholesterol-fed group and aspirin treated group, respectively. The maximum plaque thickness, the degree of artery stenosis and the proportion of the intimal circumference occupied by atheroma of the 3 groups were significantly different from each other (P&lt;0.01). The expression of COX-2 and macrophage in plaque of the aspirin treated group were decreased compared with that in untreated cholesterol-fed group. However, no difference was found in the expression of VSMC between the aspirin treated and the untreated cholesterol-fed group. Conclusion The mechanism of atherosclerosis suppression by aspirin in cholesterol-fed rabbits is related to the inhibition of COX-2 expression together with the reduced inflammation followed by, but not related to the hypolipidemic effects.     

环氧合酶-2蛋白在胃癌发生发展中的表达

目的研究环氧合酶-2（COX-2）蛋白在胃癌、癌前病变、淋巴结转移灶等组织中的表达。方法应用免疫组化SP法和RT—PCR法检测甘肃省河西地区90例胃癌,及其80例增生性病变,22例正常胃黏膜上皮和29例淋巴结转移灶中COX-2的表达。结果免疫组化结果提示正常胃黏膜上皮、增生病变及原位癌中均未发现有COX-2的表达;无淋巴结转移的胃癌原发灶中COX-2的阳性表达率是46％,有淋巴结转移的胃癌原发灶中阳性表达率是72％,淋巴结转移灶中阳性表达率是90％。在鳞状细胞浸润癌中COX-2呈低表达（14％）,在浸润性腺癌中高表达（78％）。RT-PCR结果与免疫组化结果相似。结论COX-2的表达与胃癌的浸润、淋巴结转移有关,其表达具有组织选择性。