子宫内膜癌组织COX-2和VEGF及KDR蛋白表达临床意义的探讨

目的:分析子宫内膜癌组织COX-2、VEGF和KDR蛋白的表达及临床意义.方法:应用免疫组化法对112例子宫内膜癌组织COX-2、VEGF和KDR蛋白进行检测.结果:112例子宫内膜癌组织COX-2、VEGF和KDR蛋白的阳性率分别为47.32%、66.07%和52.68%,三者表达具有协同性.Ⅱ+Ⅲ+Ⅳ期子宫内膜癌组织COX-2、VEGF和KDR蛋白表达阳性率分别为68%、88%和72%,高于Ⅰ期41.38%、59.77%和47.13%,P<0.05.低分化子宫内膜癌组织COX-2和VEGF蛋白表达阳性率100%,高于中分化61.11%和69.44%以及高分化的29.69%和57.81%,P<0.05.肌层浸润＞1/2子宫内膜癌组织COX-2、VEGF和KDR蛋白表达阳性率62.5%、96.88%和84.38%,高于肌层浸润≤1/2者41.25%、53.75%和40%,P<0.05.结论:COX-2、VEGF和KDR蛋白在子宫内膜癌织中的表达水平均较高且具有协同性.COX-2、VEGF及KDR是参与肿瘤发生、发展和转移的重要基因.     

COX-2与子宫内膜癌的关系

目的 研究子宫内膜癌中环氧合酶-2（cyclooxygenase-2，COX-2）的表达及其与临床病理因素的关系。探讨COX-2在子宫内膜癌发生、发展中的作用。方法 采用免疫组织化学S-P法检测20例正常子宫内膜、20例不典型增生子宫内膜、45例子宫内膜癌等3组标本的COX-2蛋白表达水平。结果 正常子宫内膜、不典型增生内膜及子宫内膜癌组织中率分别为15.0％（3／20）、50.0％（10／20）、77．8％（35／45），由正常子宫内膜至子宫内膜癌的进展过程中，COX-2的阳性表达率呈现逐渐增高的趋势。COX-2阳性表达率与子宫内膜癌的分期、淋巴结转移、肌层浸润深度、病理类型无明显相关性。高分化内膜癌COX-2阳性表达率（89.3％）高于中低分化内膜癌（58．8％），P〈0．05。结论 COX-2在子宫内膜癌发生、发展中起着重要的作用。COX-2表达上调可能是子宫内膜癌形成中的早期事件。     

COX-2及NF-κB p65在子宫内膜癌组织中的表达及临床意义

目的 探讨COX-2及NF-κB p65在子宫内膜癌组织中的表达及临床意义.方法 选择正常子宫内膜组织者60例(对照组),子宫内膜癌60例(观察组),两组都进行COX-2及NF-κB p65的表达检测,同时进行了相关性分析.结果 对照组COX-2阳性表达率为3.3%,观察组为55.0%.对照组NF-κB p65阳性表达率为23.3%,观察组为63.3%,对比差异都有统计学意义(P<0.05).在观察组中,随着子宫内膜癌的病理分期增加、分化程度的减少、淋巴结转移的发生,COX-2、NF-κB p65的阳性表达率都呈现增高的趋势,对比差异都有统计学意义(P<0.05).Spearman等级相关分析结果显示COX-2与NF-κB p65的表达呈正相关(γ=0.307,P<0.05).结论 COX-2、NF-κB p65在子宫内膜癌中呈现高表达状况,且分化程度低、分期晚、发生转移者高于分化程度高、分期早、未发生转移者,两者在子宫内膜癌发生发展中起共同促进作用.     

NF-κB、COX-2、LKB1在子宫内膜癌组织中的表达及临床意义

目的 探讨核转录因子κB(NF-κB)、环氧化酶2(COX-2)、肝激酶基因B1(LKB1)在不同子宫内膜组织中的表达及临床意义.方法 采用免疫组化SP法检测60例正常子宫内膜、31例非典型增生子宫内膜和60例子宫内膜癌组织中NF-κB、COX-2、LKB1蛋白的表达,并分析其与子宫内膜癌临床病理特征的关系.结果 NF-κB、COX-2、LKB1在子宫内膜癌组织、非典型增生子宫内膜组织和正常子宫内膜组织中的阳性表达率分别为63.3％、70.0％、35.0％,41.9％、64.5％、61.3％和35.0％、51.7％、93.3％,NF-κB、LKB1在3组间的表达差异有统计学意义(P＜0.05),而COX-2的表达差异无统计学意义(P＞0.05).子宫内膜癌中NF-κB、COX-2、LKB1蛋白的阳性表达与病理类型、FIGO分期、淋巴结转移均无关(P＞0.05).LKB1蛋白表达与肌层浸润深度和组织学分级相关(P＜0.05),NF-κB蛋白表达与组织学分级亦相关(P＜0.05).结论 NF-κB在子宫内膜癌组织中高表达、LKB1低表达,其表达可能与子宫内膜癌的发生发展有关.     

环氧合酶-2 在子宫内膜癌中的表达及其与肿瘤血管形成的关系

 目的 探讨环氧合酶-2(Cyclooxygenase-2，COX2)在子宫内膜癌组织中表达及与肿瘤血管形成的关系。方法 采用免疫组化SIP方法检测34例子宫内膜癌组织COX-2和血管内皮生长因子(Vascu1arendothelial growth factor，VEGF)的表达和微血管密度(Microvessel density，MVD)，观察COX-2表达与肿瘤血管形成之间的相关性。结果 COX-2在子宫内膜癌组织中的阳性表达率为64．7％，而对照组正常子宫内膜均未见表达，内膜癌组的中分化细胞COX-2蛋白的表达高于低分化细胞，差异有显著性(P<0．05)；COX-2表达阳性组和阴性组MVD分别为(41．53±19．10和28．79±8．20)，两组比较具有显著性差异(P<0．05)。COX-2的表达评分与VEGF及MVD高度均呈正相关(P<0．01；P<0．0001)。结论 COX-2可能主要参与子宫内膜癌发生的早期；子宫内膜癌组织中COX-2的高表达可能在VEGF诱导肿瘤血管形成的过程中起重要作用。     

COX-2、VEGF、Ki67在子宫内膜癌中的表达及意义

目的探讨环氧合酶-2(COX-2)、血管内皮生长因子(VEGF)和增殖细胞核相关抗原(Ki67)在子宫内膜癌发生、发展中的作用。方法采用免疫组织化学Envision法检测50例子宫内膜癌和15例正常子宫内膜组织中COX-2、VEGF、Ki67的表达。结果子宫内膜癌组织中COX-2、VEGF、Ki67的表达明显高于正常子宫内膜组织(P<0.01);COX-2的高表达与临床分期无关(P>0.05),与分化程度和浸润深度有关(P<0.01);VEGF的高表达与临床分期、浸润深度无关(P>0.05),分化程度有关(P<0.05);Ki67的高表达与临床分期(P<0.05)、分化程度、浸润深度有关(P<0.01)。COX-2与VEGF和Ki67的表达间存在正相关(r=0.39,P<0.01;r=0.32,P<0.05)。结论COX-2、VEGF、Ki67与子宫内膜癌的发生和发展相关,COX-2可通过使VEGF高表达,增加细胞增殖活性促进子宫内膜癌的发生、发展。     

人子宫内膜癌组织COX-2和CD44V6及Bcl-2表达临床意义分析

目的:探讨人子宫内膜癌组织中环氧化酶-2(cyclooxygenase-2,COX-2)、CD44V6及Bcl-2的表达状况及其临床意义。方法:使用流式细胞仪(flow cytometry,FCM)检测55例人子宫内膜癌组织及36例癌旁组织中COX-2、CD44V6及Bcl-2蛋白表达。结果:子宫内膜癌组织中COX-2、CD44V6和Bcl-2蛋白的表达率分别为(39.06±18.51)%、(36.73±2.97)%和(12.52±5.23)%,癌旁组织中分别为(2.85±1.42)%、(2.93±1.78)%和(3.07±1.66)%。三者在癌组织中的表达明显高于癌旁组织,差异有统计学意义,P<0.05。COX-2及CD44V6表达与人子宫内膜癌患者的年龄(P=0.08)、病理分级(P=0.11)无关,而与肿瘤的分期及淋巴结转移有关,P<0.05。Bcl-2蛋白表达与患者年龄、病理分级、临床分期及淋巴结转移均无关,P>0.05。COX-2与CD44V6蛋白表达正相关,r=0.507,P<0.01;COX-2及CD44V6与Bcl-2蛋白表达无关。结论:人子宫内膜癌组织中COX-2和CD44V6高表达的患者更易发生浸润、转移,且两者有正协同作用。Bcl-2与人子宫内膜癌的发生有一定关系。     

子宫内膜癌手术预后因素的多因素分析

目的:分析和评价子宫内膜癌常用临床病理因素(绝经前后、手术病理分期、组织学分级、淋巴结转移、病理类型、术后辅助治疗、肌层浸润深度)和生物学因素(COX-2、VEGF、KDR及ER、PR)对子宫内膜癌手术预后的影响,筛选出对子宫内膜癌手术预后最有显著的影响因素.方法:将可能与子宫内膜癌预后有关的常用临床病理因素和生物学因素进行单因素分析.通过Cox模型对单因素分析筛选出的影响子宫内膜癌预后的因素进行多因素分析.结果:137例子宫内膜癌患者中,通过随访,共获得确切随访者112例,5年生存率为83.04%.Ⅰ～Ⅳ期5年生存率分别为89.66%,80.00%,53.85%,0.子宫内膜癌组织中COX-2、VEGF及KDR蛋白的阳性率分别为47.32%、66.07%及52.68%,且三者表达具有协同性.COX-2、VEGF、KDR蛋白阳性患者5年累积生存率分别为75.47%、78.38%、72.88%.将这些临床病理因素及生物学因素进行单因素分析,结果手术病理分期、组织学分级、肌层浸润深度、有无淋巴结转移、COX-2阳性表达、KDR阳性表达、ER阳性表达、PR阳性表达8个因素与预后明显相关(P 均＞＜0.05＝,绝经前后、病理类型、术后辅助治疗、VEGF阳性表达与预后无关(P 均＞0.05).应用Cox模型对单因素分析筛选出的8个有显著意义的因素进行多因素分析表明:肌层浸润深度、手术病理分期、组织学分级是影响子宫内膜癌手术预后最显著的3个因素.结论:肌层浸润深度、手术病理分期、组织学分级是影响子宫内膜癌手术预后的因素,但浸润肌层是影响子宫内膜癌手术预后的最重要的因素.     

塞来昔布对子宫内膜癌细胞增生和COX-2 mRNA表达的影响

 【目的 探讨塞来昔布对子宫内膜癌细胞HEC-1-B的作用及对COX-2 mRNA表达的影响。方法 采用细胞培养、MTT试验研究不同浓度的塞来昔布对人类子宫内膜癌细胞HEC-1-B生长的抑制作用，通过RT-PCR法检测塞来昔布作用后COX-2 mRNA表达的变化。结果 塞来昔布能够有效地抑制HEC-1-B细胞的生长，并呈一定的剂量和时间依赖关系，同时能够抑制其COX-2 mRNA表达，20 μmol／L和50 μmol／L塞来昔布对COX-2 mRNA表达的抑制率分别为25.92 ％和50.81％。结论 塞来昔布可能是通过降低COX-2 mRNA的表达而抑制子宫内膜癌细胞的生长。     

环氧合酶-2在子宫内膜癌中表达及临床意义的研究进展

环氧合酶-2（cox-2）是前列腺素合成过程中的重要限速酶，研究表明其过度表达与肿瘤的发生发展密切相关，可能通过促进细胞增殖，抑制细胞凋亡，促进血管形成及抑制机体免疫等机制促进肿瘤的形成。近年来研究发现COX-2不仅在子宫内膜癌组织中高表达，并与子宫内膜癌的一些临床病理特征和预后之间存在联系，提示其可能成为判断肿瘤恶性程度、转移能力及预后的新的生物学指标。现将COX-2在子宫内膜癌中的表达及其临床意义的研究进展作一综述。     

Cyclooxygenase-2 expression in endometrial carcinoma: correlation with clinicopathologic parameters and clinical outcome

BACKGROUND: Cyclooxygenase-2 (COX-2) is overexpressed in endometrial hyperplasia and carcinoma, but no data have been reported until now about the expression of COX-2 and its possible clinical significance in endometrial carcinoma. We investigated by immunohistochemistry the expression of COX-2 in a single institutional series of primary untreated endometrial carcinoma patients. The relationship between COX-2 expression and microsatellite instability (MI) status was also analyzed. METHODS: The study was conducted on 69 primary untreated endometrial carcinoma patients who were admitted to the Department of Obstetrics and Gynecology, Catholic University of Rome. Immunohistochemistry was performed by using rabbit polyclonal antiserum against human COX-2. Analysis of MI was performed for 47 patients with endometrial carcinomas. RESULTS: Twenty-seven patients (39.1%) were scored as COX-2 positive. COX-2 positivity was higher (60.8%) in endometrial carcinoma with cervical or extrauterine involvement than in tumors limited to the corpus (28.3%; P = 0.0174). COX-2 positivity increased from Grade 1 (13.6%) to Grade 2 (41.7%) to Grade 3 (60.9%) endometrial carcinoma (P = 0.0049). Interestingly, considering early International Federation of Gynecology and Obstetrics stage patients (n = 53), the percentage of COX-2 positivity was higher in patients with deep myometrial invasion (66.7%) than in patients without or less than 50% myometrial invasion (15.6%) (P = 0.0003). No association between COX-2 and MI status was found. COX-2-positive patients showed a trend to a shorter disease-free survival than COX-2-negative patients (P = 0.09). CONCLUSIONS: COX-2 is expressed in a high percentage of a large series of primary endometrial tumors and its expression may be associated closely with parameters of tumor aggressiveness The possible prognostic role of COX-2 in endometrial carcinoma deserves further study.     

子宫内膜癌组织中环氧化酶-2表达及其与血管生成的关系

目的　研究环氧化酶 2 (COX 2 )在子宫内膜癌组织中的表达及其与血管生成的关系。方法　用免疫组化法检测42例子宫内膜癌和15例正常子宫内膜组织中COX 2的表达,并检测微血管密度(MVD)。结果　子宫内膜癌组织中COX 2、MVD明显高于正常子宫内膜组织(P <0 0 5 ) ,COX 2高表达与子宫内膜癌组织学分级和浸润深度有关(P <0 0 5 ) ,与临床分期、淋巴结转移和组织学分型无关(P >0 0 5 ) ,COX 2表达阳性者MVD明显高于COX 2表达阴性者(P <0 0 5 )。结论　COX 2过表达可能参与子宫内膜癌发生发展过程,且与子宫内膜癌新生血管形成有关。     

Role of cyclooxygenase-2 in immunomodulation and prognosis of endometrial carcinoma

Although there are several hypotheses explaining the mechanisms of immune-privileged status of malignant tumor, the exact pathway has yet to be explored. Cyclooxygenase (COX)-2 plays a vital role in prognosis of cancer patients in terms of contribution to neoangiogenesis and apoptosis inhibition; however, the impact of COX-2 in immunomodulation has not been reported. We have evaluated the expression of COX-2 and its impact on infiltration of immune-competent cells into the tumor cell nest in endometrial carcinoma. Tissue specimens from 70 endometrial carcinoma patients who had undergone a curative resection were evaluated for COX-2 expression and host immune status (CD8+ T cells). COX-2 expression was associated with FIGO stage and myometrial invasion, but there was no statistically significant impact. CD8+ T cells within cancer cell nest (Nest CD8) were inversely correlated with the expression level of COX-2 (p = 0.0006). Nest CD8 became an independent predictor of patient survival (Hazard ratio = 10.300, p = 0.0304) in Cox's multivariate analysis. The expression level of COX-2 was found to be a significant predictor of disease relapse in univariate analysis (p = 0.0294) but not in multivariate analysis (p = 0.5949). In conclusion, increased nest CD8 produced a survival advantage in endometrial carcinoma patients. Moreover, tumor-produced COX-2, which reduces the infiltration of CD8+ T cells into cancer cell nests, may allow tumors to avoid immune surveillance.     

Effects of ectopic HER-2/neu gene expression on the COX-2/PGE2/P450arom signaling pathway in endometrial carcinoma cells: HER-2/neu gene expression in endometrial carcinoma cells

Objectives

To investigate the role of HER-2/neu-mediated COX-2/P450arom signal in estrogen-dependent endometrial carcinoma.

Methods

The recombinant eukaryotic expression vector, pcDNA3.1-HER-2/neu, was constructed and transfect to Ishikawa endometrial carcinoma cells. The expression of COX-2 and P450arom in transfected cells were detected by real-time PCR and western blotting. The levels of estrogen in cell supernatants were detected by ELISA.

Results

Over-expression of HER-2/neu in transfected cells was confirmed by real-time PCR and western blotting. The levels of autocrine estrogen in transfected cells was significantly increased which combination with the enhancement of COX-2 and P450arom expression in transfected cells.

Conclusion

HER-2/neu induced the improvement of autocrine estrogen in endometrial carcinoma cell through triggering the COX-2/P450arom signal.     

HER-2/neu COX-2 PGE2 P450arom在子宫内膜癌组织的表达及其相关性研究

探讨子宫内膜腺癌组织中HER-2/neu、COX-2、PGE2、P450arom 4种生物学指标的表达,分析这4种生物学指标表达强度之间相关性及与临床病理因素的相关性。方法:应用逆转录-聚合酶链反应（RT-PCR）技术及酶联免疫吸附试验（ELISA）检测HER-2/neu、COX-2、P450arom及PGE2在正常子宫内膜、子宫内膜不典型增生和子宫内膜腺癌组织中的表达情况。结果:HER-2/neu、COX-2、P450arom及PGE2在癌组织中表达高于非癌组织,有显著性差异（P<0.01）。癌组织中HER-2/neu与COX-2、P450arom表达强度呈正相关（r值分别为0.931,0.934,P<0.05）,COX-2与P450arom表达强度亦呈正相关（r=0.908,P<0.05）。COX-2mRNA和PGE2表达强度变化呈正相关（r=0.552,P<0.05）。结论:HER-2/neu、COX-2、P450arom及PGE2可能存在协同作用,共同促进子宫内膜腺癌的发生发展。      

Expression of cyclooxygenase-2 in uterine endometrial cancer and anti-tumor effects of a selective COX-2 inhibitor

A role for cyclooxygenase-2 (COX-2) in the development and progression of various tumors has been identified. Selective COX-2 inhibitors produce anti-proliferative effects in various cancer cell lines that express COX-2. However, the mechanisms underlying anti-tumor effects are unclear. Furthermore, few studies have studied COX-2 expression in gynecological cancers, especially endometrial cancer. The current study had two goals. We investigated the correlation between COX-2 expression and clinicopathological factors of uterine endometrial cancer. We also investigated effects of treatment with etodolac, a selective COX-2 inhibitor, on the uterine endometrial cancer cell line TMG-L, which expresses COX-2. We conclusively confirmed expression of COX-2 mRNA and protein in endometrial cancer that exceeded levels of COX-2 seen in normal endometrium. However, no significant correlations were observed between COX-2 expression in endometrial cancer tumor samples and clinicopathological factors or disease-free survival rate of patients with endometrial cancer. Study of COX-2 inhibition of TMG-L cells showed that etodolac produced dose-dependent inhibition of cell proliferation through G1 phase cell-cycle arrest. Etodolac-induced cell-cycle arrest might be caused by increases in p53 and P21WAF1 protein expression. Production of basic-fibroblast growth factor (bFGF, a pro-angiogenesis factor) was inhibited by etodolac in a dose-dependent manner. Furthermore, telomerase activity was inhibited and expression of hTERT mRNA was significantly inhibited with etodolac, leading to the conclusion that anti-tumor effects of etodolac on TMG-L cells are due to inhibition of both angiogenesis and telomerase activity. These results strongly suggest that COX-2 inhibitors have potential as therapeutic (and possibly, chemopreventive) agents for endometrial cancers that overexpress COX-2.     

慢病毒介导COX-2基因的shRNA对子宫内膜癌侵袭力的影响

目的：探讨慢病毒介导COX-2基因的shRNA对子宫内膜癌HEC-1B细胞侵袭力的影响。方法：设计、合成4对针对COX-2基因shRNA的寡核苷酸序列,分别构建质粒,转染293T细胞,据其对COX-2的抑制率,筛选最有效的COX-2基因RNAi靶序列,设计并合成其shRNA的双链DNA Oligo,与含绿色荧光蛋白（GFP）编码基因的pGCL-GFP线性化载体连接产生重组慢病毒质粒LV-COX-2-ShRNA,并行PCR和测序鉴定。将LV-COX-2-ShRNA,pHelper 1.0和pHelper 2.0三种质粒DNA共转染293T细胞,包装产生慢病毒,以293T细胞GFP蛋白的表达水平采用逐孔稀释滴度法测定病毒滴度。重组慢病毒感染人子宫内膜癌HEC-1B细胞,行RT-PCR和western-blot验证COX-2基因的沉默效果,用Transwell侵袭试验检测HEC-1B细胞体外侵袭力的改变。结果：成功构建COX-2基因RNA干扰慢病毒载体并获得相应的慢病毒,浓缩病毒悬液的滴度为5×107Tu/ml。和对照组相比,RNA干扰组HEC-1B细胞COX-2基因的mRNA水平和蛋白水平的抑制率分别为61.87%和67.48%;其穿过人工基底膜的平均数为16.6,明显少于空白对照组50.2及非特异性siRNA感染组47.2,侵袭抑制率为64.8%（P〈0.01）。结论：RNA干扰可有效抑制HEC-1B细胞的侵袭能力。