环氧合酶-2及其抑制剂与子宫内膜癌

环氧合酶-2(COX-2)参与子宫内膜癌的发生、发展,可能影响其预后.COX-2在子宫内膜癌中的表达诱导肿瘤细胞增殖,抑制细胞凋亡,促进肿瘤细胞侵袭和转移.因此,COX-2抑制剂有望成为子宫内膜癌新的防治方法.     

同源盒基因MSX2在子宫内膜癌中的表达和功能研究

目的;通过检测 ⅡI类肌节同源盒基因2（muscle segment homeobox 2，MSX2）在子宫内膜癌中的表达情况.分析其表达与临床病理参数的关系，并初步探索 MSX2 基因在子宫内膜癌细胞中的生物学功能。方法∶通过生物信息学、Real-Time PCR 及免疫组化方法分析 MSX2 基因在正常子宫内膜及子宫内膜癌中的表达及与临床病理，预后的关系。使用 CCK8 实验、细胞划痕实验 、Transwell 实验验证 MSX2 基因的表达调控子宫内膜癌细胞的增殖、侵袭与转移。结果;MSX2 基因在子宫内膜癌组织中旱现高表达，肿瘤分级越低，FIGO 分期越早，MSX2 表达越高.高表达 MSX2 的患者预后更好。MSX2 基因的高表达能够抑制子宫内膜癌细胞增殖、侵袭和转移能力。结论∶MSX2 基因可能在子宫内膜癌中发挥了抑制肿瘤进展的功能。     

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。     

PDCD5在人子宫内膜癌中的表达及对细胞凋亡的影响

目的 探讨PDCD5对子宫内膜癌细胞克隆形成,增殖以及凋亡的作用.方法 收集行手术切除的子宫内膜癌及对应癌旁组织共60例,采用qRT-PCR,western blot分别检测PDCD5 mRNA水平及蛋白水平在子宫内膜癌及对应癌旁组织中的表达情况,通过重组PDCD5(rhPDCD5)处理HEC1A子宫内膜癌细胞,检测其对细胞增殖、克隆形成及细胞凋亡的影响.结果 子宫内膜癌组织中PDCD5表达水平显著低于对应癌旁组织(P＜0.05);肿瘤组织中PDCD5低表达与病理分期、组织学分级、肌层浸润以及肿瘤淋巴结转移均具有显著相关性(P＜0.05);低表达PDCD5的患者总生存时间均显著缩短(P＜0.05);重组的PDCD5蛋白能够通过促进子宫内膜癌细胞凋亡抑制细胞增殖和克隆形成能力.结论 PDCD5在子宫内膜癌组织中低表达并与子宫内膜癌临床病理特征及不良预后显著相关,并能够通过促进子宫内膜癌细胞凋亡进而抑制细胞增殖及克隆形成,PDCD可能参与子宫内膜癌发生发展进程,并可作为早期诊断及预后评价的重要分子.     

同源盒基因MSX2在子宫内膜癌中的表达和功能研究

目的;通过检测 ⅡI类肌节同源盒基因2（muscle segment homeobox 2,MSX2）在子宫内膜癌中的表达情况.分析其表达与临床病理参数的关系,并初步探索 MSX2 基因在子宫内膜癌细胞中的生物学功能。方法∶通过生物信息学、Real-Time PCR 及免疫组化方法分析 MSX2 基因在正常子宫内膜及子宫内膜癌中的表达及与临床病理,预后的关系。使用 CCK8 实验、细胞划痕实验 、Transwell 实验验证 MSX2 基因的表达调控子宫内膜癌细胞的增殖、侵袭与转移。结果;MSX2 基因在子宫内膜癌组织中旱现高表达,肿瘤分级越低,FIGO 分期越早,MSX2 表达越高.高表达 MSX2 的患者预后更好。MSX2 基因的高表达能够抑制子宫内膜癌细胞增殖、侵袭和转移能力。结论∶MSX2 基因可能在子宫内膜癌中发挥了抑制肿瘤进展的功能。     

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。     

bcl-2、Survivin 在子宫内膜癌组织中的表达及其意义

近年来子宫内膜癌发病率逐渐上升。研究显示，肿瘤的发生发展与细胞凋亡障碍关系更为密切。很多研究还证明bcl-2是调节细胞凋亡的关键性生物因子。新近发现的凋亡抑制基因Survivin属于凋亡抑制蛋白家族的新成员。对于bcl-2及Survivin蛋白在子宫内膜癌中表达的研究少见报道。我们采用免疫组织化学SP法研究bcl-2及Survivin蛋白在子宫内膜癌中的表达情况，旨在探讨其在子宫内膜癌发生发展过程中的意义。     

子宫内膜癌组织中PRMT2高表达及其促进细胞增殖的机制探讨

目的:探究蛋白质精氨酸甲基转移酶2（protein arginine methyltransferase 2，PRMT2）在子宫内膜癌恶性进展中的作用及临床意义。方法:免疫组化方法检测子宫内膜癌组织及其癌旁组织中PRMT2的表达。通过感染PRMT2 shRNA慢病毒和转染pcDNA3.1-PRMT2质粒，构建子宫内膜癌HEC-1A细胞PRMT2敲低和过表达模型，CCK-8和克隆形成实验检测PRMT2对细胞增殖的影响。通过RT-qPCR探明PRMT2下游靶基因，并采用Western blot实验进行验证。采用挽救实验证实PRMT2调控靶基因的特异性和在子宫内膜癌增殖中的重要性。结果:免疫组化实验发现PRMT2在子宫内膜癌组织中表达明显高于癌旁组织，且PRMT2表达与患者生存期呈显著负相关（P＜0.05）。敲低PRMT2表达能显著抑制子宫内膜癌细胞增殖（P＜0.05），并且发现PRMT2敲低可以抑制CCND1表达；相反，过表达PRMT2能够促进子宫内膜癌细胞增殖（P＜0.05），促进CCND1表达。挽救实验结果表明，在PRMT2敲低的细胞中过表达CCND1能够部分恢复细胞的增殖能力。结论:PRMT2在子宫内膜癌组织中高表达，并且PRMT2能够通过调控CCND1表达促进子宫内膜癌细胞的增殖。     

环氧化酶-2表达与肿瘤关系的研究进展

环氧化酶-2(COX-2)在多种恶性肿瘤中高表达。通过抑制细胞凋亡等机理促进肿瘤的发生和发展。本文综述了COX-2基因结构、生物学特点和在常见恶性肿瘤中的表达及预后的研究进展。     

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.     

Akt regulates COX-2 mRNA and protein expression in mutated-PTEN human endometrial cancer cells

In human endometrial cancer, the fourth most common cancer in women, tumor suppressor phosphatase tensin homologue (PTEN) is frequently mutated. In the presence of a mutated PTEN protein, Akt phosphorylation levels are increased leading to the activation of this survival pathway. Numerous studies indicated that COX-2 is inappropriately induced and up-regulated in a number of malignant cancer cells. COX-2 plays an important role in tumor cell biology, taking part actively in angiogenesis particularly via the production of prostaglandin E2 (PGE2). The present study was undertaken to determine the involvement of PI 3-K/Akt pathway in the regulation of COXs expression and PGE2 synthesis. Three different human endometrial cancer cell lines known to have wild-type PTEN (HEC 1-A) or a mutated inactive PTEN protein (RL 95-2 and Ishikawa) were used for these studies. Results showed that Akt phosphorylation was high in mutated PTEN cells. RT-PCR studies revealed that Akt1 and Akt2 were the regulated forms whereas Akt3 mRNA was nearly undetectable. COX-2 mRNA expression and protein levels were high in these cells compared to wild-type PTEN cells as demonstrated by RT-PCR and Western analysis respectively. PGE2 production was higher in mutated-PTEN expressing phospho-Akt and COX-2 compared to wild-type PTEN cells. Inhibition of PI 3-K with Wortmannin and LY294002 blocked Akt phosphorylation and inhibited expression of COX-2 in mutated-PTEN cells. Inhibition of Akt phosphorylation with specific PI 3-K inhibitors and down-regulation of COX-2 increased apoptosis in human endometrial cancer cells. Likewise, transfection of mutated-PTEN cells with a dominant negative Akt vector, resulted in COX-2 down-regulation and activation of apoptosis, as demonstrated by Hoechst nuclear staining. On the opposite, activation of Akt using a constitutively active expression vector, resulted in the up-regulation of COX-2 protein expression. Specific inhibition of COX-2 with NS-398 induced apoptosis in COX-2 expressing human endometrial cancer cells. It is concluded that the PI 3-K/Akt survival pathway is involved in the regulation of COX-2 and PGE2 synthesis in human endometrial cancer cells.     

Cyclooxygenase-2 and p53 expressions in endometrial cancer.

Cyclooxygenase-2 (COX-2) has been known to be related with various types of carcinoma, but we have insufficient knowledge about the association between COX-2 and endometrial cancer. Many have reported a close relationship between p53 expression and a poor prognosis in endometrial cancer, but it is unclear whether p53 is an independent prognostic factor. To clarify these uncertainties, we examined the expressions of COX-2 and p53 in endometrial cancer tissues. The study was carried on 152 endometrial cancer patients who had operation at Seoul National University Hospital. Paraffin-embedded tissue blocks were sectioned and immunostained using monoclonal anti-COX-2 and anti-p53 antibodies. Twenty-seven (17.8%) specimens stained as COX-2 positive. COX-2 positivity was more frequently observed in postmenopausal patients than in premenopausal patients (8.8% versus 25.0%; P = 0.009). However, COX-2 positivity did not show a statistically significant association with any other clinicopathologic characteristic (parity, body mass index, histotype, International Federation of Gynecology and Obstetrics stage, grade, lymph node metastasis, deep myometrial invasion, or p53 overexpression). Thirty-one (20.4%) specimens showed p53 overexpression and this was significantly correlated with an advanced stage (P = 0.001), poor differentiation (P < 0.001), lymph node metastasis (P = 0.012), and deep myometrial invasion (P < 0.001). Multivariate Cox regression analysis showed that advanced stage was an independent prognostic factor of survival, but p53 overexpression was not. COX-2 may be associated with endometrial cancer carcinogenesis during the postmenopausal period but not with tumor aggressiveness and p53 overexpression. The p53 overexpression was found to be strongly associated with endometrial cancer aggressiveness.     

Hepatocyte growth factor induces anoikis resistance by up-regulation of cyclooxygenase-2 expression in uterine endometrial cancer cells

Cyclooxygenase-2 (COX-2) has been implicated in the promotion of carcinogenesis. Although the role of COX-2 in endometrial cancer remains unclear, recent experiments suggest that COX-2 antagonizes cell apoptosis, increases the invasiveness of malignant cells, and promotes angiogenesis. Hepatocyte growth factor (HGF) is a mesenchymal-derived cytokine and the interaction between HGF and its tyrosine kinase receptor, c-Met proto-oncogene, is associated with tumor progression and metastasis. To investigate the molecular mechanism of HGF-induced anoikis resistance, we analyzed the signal transduction and COX-2 expression in endometrial cancer cells. Here, we show i) the expression of COX-2 protein significantly increased in a dose-dependent manner after HGF stimulation in endometrial cancer cell lines (HEC-IB and RL95-2), reaching 200-270% stimulation at the highest doses of HGF tested (40 ng/ml); ii) flow cytometry and TUNEL analyses revealed that HGF significantly inhibited anoikis of RL95-2 cells; iii) phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), but not mitogen-activated protein kinase/ERK kinase (MEK) inhibitor (PD98059), specifically blocked HGF-mediated anoikis resistance in RL95-2 cells; and iv) COX-2 inhibitor, Meloxicam, abrogated HGF-mediated anoikis resistance. Our data suggest that HGF induces anoikis resistance in endometrial cancer cells possibly through PI3K/Akt pathway-dependent up-regulation of COX-2 expression.     

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.     

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.     

Non-steroidal anti-inflammatory drugs inhibit cellular proliferation and upregulate cyclooxygenase-2 protein expression in endometrial cancer cells

We determined the effects of several non-steroidal anti-inflammatory drugs (NSAIDs), aspirin (acetylsalicylic acid, ASA), indomethacin and a cyclooxygenase-2 (COX-2)-selective inhibitor (NS398), on cellular proliferation and regulation of COX-2 protein expression in endometrial cancer cells in vitro, and investigated their modes of action. All three NSAIDs markedly inhibited the proliferation of Ishikawa, HEC-1A and AN3CA endometrial cancer cell lines in a time- and concentration-dependent manner. ASA and indomethacin triggered apoptosis in cells of all three lines through release of cytosolic cytochrome c, activation of caspase-9 and-3, and cleavage of poly(ADP-ribose) polymerase (PARP), but NS398 induced minimal apoptosis only in Ishikawa cells. ASA altered the cell cycle distribution, with G2/M phase accumulation of cells, and induced overexpression of Ki-67 protein. Both ASA and indomethacin reduced the protein levels of Bcl-2 and Bcl-xl, but upregulated those of Bax and Bcl-xs. COX-2 protein expression and PGE(2) production were upregulated by ASA and indomethacin in all three cell lines. However, NS398 did not alter COX-2 protein expression or PGE(2) production in these cells. These results indicate that NSAIDs inhibit proliferation of endometrial cancer cells independently of the reduction of COX-2 protein expression. A cytochrome c-dependent apoptotic pathway and/or cell cycle arrest may contribute to the inhibitory effects of these NSAIDs.     

环氧合酶-2及诱导型一氧化氮合酶在子宫内膜癌组织中的表达及意义

 目的　探讨环氧合酶 2 (COX 2 )与诱导型一氧化氮合酶 (iNOS)在子宫内膜癌组织中的表达及与子宫内膜癌发生和进展的关系。方法　应用免疫组化方法检测 30例子宫内膜癌组织中COX 2和i NOS的表达 ;并以逆转录聚合酶链反应 (RT -PCR)技术检测COX 2及iNOSmRNA的表达。结果　子宫内膜癌组织中COX 2及iNOS蛋白阳性表达率分别为 6 6 .7%和 73.3% ;子宫内膜癌及癌旁组织中COX 2及iNOSmRNA的表达均明显上调。中分化内膜癌组织COX 2蛋白的表达显著高于低分化的癌组织 (P <0 .0 5 )。子宫肌层浸润深度 >1/2的iNOS表达明显高于肌层未受累或≤ 1/2的患者 30例 (P<0 .0 5 )。COX 2和iNOS蛋白的表达呈明显的正相关 (r =0 .6 0 1,P <0 .0 0 1)。结论　COX 2和iNOS的高表达与子宫内膜癌的发生及发展密切相关 ;COX 2可能主要参与子宫内膜癌发生的早期 ;iNOS的高表达与子宫内膜癌浸润能力有关