胃癌组织中环氧合酶-2表达与其临床病理因素及催化产物的关系

目的检测环氧合酶-2(Cyclooxygenase-2,COX-2)蛋白在胃癌组织中的表达,探讨COX-2表达与胃癌临床病理因素及其催化产物PGE2、TXA2的关系。方法应用免疫组织化学技术及放射免疫分析法检测胃癌组织中COX-2蛋白表达及PGE2、TXA2含量。结果COX-2在62.2％胃癌组织中表达增高,COX-2表达与胃癌病灶大小、淋巴结转移和TNM分期密切相关(P<0.05)。COX-2阳性表达的胃癌组织中PGE2水平明显升高,而TXB2水平在各组间差异均无显著性。结论 胃癌组织中COX-2表达增加与肿瘤大小、淋巴结转移、TNM分期有关,COX-2表达与其产物PGE2水平密切相关,COX-2可能通过催化PGE2的合成而在胃癌形成中起作用。     

Expression of cyclooxygenase-2, Bcl-2 and Ki-67 in pleomorphic adenoma with special reference to tumor proliferation and apoptosis

The present report is of immunohistochemical and cell biological studies on cyclooxygenase(COX)-2, Bcl-2 and Ki-67 in pleomorphic adenoma (n = 35). Pleomorphic adenomas from oral minor glands (n = 15), parotid gland (n = 11), Submandibular gland (n = 8) were studied and sublingual gland (n = 1). Expression of COX-2, Bcl-2 and Ki-67 was examined immunohistologically. Immunostaining intensity of COX-2 and Bcl-2 was classified into expression levels from +3 to 0. The positivity to Ki-67 was evaluated by counting the number of positive cells per 1000 cells, and the values were expressed in as percentage. Apoptotic cells were detected using the modified TUNEL method. Expression of COX-2 mRNA was analyzed by real time PCR using fresh tissues of 4 cases. The relationship between morphological findings and the level of COX-2 mRNA expression was analyzed using a laser capture microdissection (LCM). The results of immunohistochemistry and gene analysis using LCM revealed expression of COX-2 mainly in luminar tumor cells. Expression of COX-2 in pleomorphic adenoma was correlated with expression of Bcl-2 statistically (p = 0.044 < 0.05, r = 0.342). There was only a small number of apoptotic cells cells, and intensity of expression of TUNEL was not correlated with the expression of COX-2 (p = 0.463 > 0.05, r = -0.128). There was no statistical correlation between COX-2 and Ki-67 (p = 0.97 > 0.05, r = -0.07). It is suggested that COX-2 may inhibit apoptosis mediating Bcl-2 expression in pleomorphic adenoma, rather than playing a role in cell proliferation.     

乳腺癌组织中环氧化酶-2 mRNA的表达及其临床病理意义

目的：探讨环氧化酶－2（COX－2）mRNA在乳腺癌中的表达以及与乳腺癌临床病理特征的关系。方法：以β-actin基因为参照,应用逆转录聚合酶链反应（RT－PCR）技术,检测30例乳腺癌组织及癌旁正常腺体中COX－2 mRNA的表达。应用条带密度分析软件,定量分析RT－PCR产物电泳带密度。结果：30例癌组织中,28例COX－2 mRNA表达水平明显增高,全距为0.05-0.91,中位数为0.53;癌旁正常腺体中无表达或表达很弱,全距为0－0.09,中位数为0。癌组织与正常腺体中COX－2的表达差异有显著性（P＜0．05）。COX－2的高表达与淋巴结的转移有关（P＜0．05）,而与年龄、癌肿的大小、临床病理分期、病理类型无关（P＞0．05）。结论：乳腺癌组织中COX－2 mRNA表达水平高于正常腺体,其表达与乳腺癌的发生、发展及淋巴结转移有关。     

Influence of cigarette smoking on prostaglandin synthesis and cyclooxygenase-2 gene expression in human urinary bladder cancer

The present study examines the effect of tobacco smoking on the expression of cyclooxygenase (COX)-2 gene, COX enzymatic activity and prostaglandin (PG) synthesis in urothelial mucosal tissues from patients with bladder cancer and from normal individuals. The detection frequency of COX-2 mRNA was 2-fold higher in bladder cancer patients compared to controls and it was accompanied by a significantly increased COX enzymatic activity and PGE2 synthesis (p < 0.05). Smokers, in both control and patients groups, had higher COX-2 expression, COX activity, and PGE2 synthesis compared to the nonsmokers (p < 0.05). The number of cigarettes smoked in the cases, but not controls, correlated well with COX enzymatic activity (r = 0.42, p = 0.016). The observed over-expression of COX-2 gene in human urinary bladder and the concomitant increases in PG synthesis may explain, at least in part, the mechanism by which cigarette smoking influences the development of urothelial neoplasia.     

Cyclooxygenase-2 pathway correlates with VEGF expression in head and neck cancer. Implications for tumor angiogenesis and metastasis

We evaluated the role of COX-2 pathway in 35 head and neck cancers (HNCs) by analyzing COX-2 expression and prostaglandin E2 (PGE2) production in relation to tumor angiogenesis and lymph node metastasis. COX-2 activity was also correlated to vascular endothelial growth factor (VEGF) mRNA and protein expression. COX-2 mRNA and protein expression was higher in tumor samples than in normal mucosa. PGE2 levels were higher in the tumor front zone in comparison with tumor core and normal mucosa (P<.0001). Specimens from patients with lymph node metastasis exhibited higher COX-2 protein expression (P=.0074), PGE2 levels (P=.0011) and microvessel density (P<.0001) than specimens from patients without metastasis. A significant correlation between COX-2 and tumor vascularization (r(s)=0.450, P=.007) as well as between COX-2 and microvessel density with VEGF expression in tumor tissues was found (r(s)=0.450, P=.007; r(s)=0.620, P=.0001, respectively). The induction of COX-2 mRNA and PGE2 synthesis by EGF and Escherichia coli lipopolysaccharide (LPS) in A-431 and SCC-9 cell lines, resulted in an increase in VEGF mRNA and protein production. Indomethacin and celecoxib reversed the EGF- and LPS-dependent COX-2, VEGF, and PGE2 increases. This study suggests a central role of COX-2 pathway in HNC angiogenesis by modulating VEGF production and indicates that COX-2 inhibitors may be useful in HNC treatment.     

Clinical implication of expression of cyclooxygenase-2 and peroxisome proliferator activated-receptor gamma in epithelial ovarian tumours

Expression of cyclooxygenase (COX)-2 plays a key role in tumorigenesis and development and peroxisome proliferator-activated receptor gamma (PPARgamma) has been implicated in the control of COX-2 expression in some tissues. The aim of this study is to investigate (1) whether expression of COX-2 and PPARgamma is associated with ovarian carcinogenesis and progression of ovarian tumours and (2) whether COX-2 expression is controlled through ligand-mediated activation of PPARgamma in ovarian carcinoma cells. For this purpose, the presence of COX-2 and PPARgamma was immunohistochemically examined in 71 epithelial ovarian carcinomas, 18 borderline tumours and 23 benign tumours and the levels of COX-2 and PPARgamma proteins were determined by enzyme immunoassay in four benign tumours, three borderline tumours and 12 carcinomas. The frequency of COX-2 and PPARgamma detection was significantly increased and decreased as lesions progressed to carcinoma, respectively. The COX-2 protein was not detected in the three borderline tumours, whereas PPARgamma protein was detected in all of them. COX-2 protein was detected in eight of the 12 carcinomas, whereas PPARgamma protein was detected in only two cases. In addition, PPARgamma protein was not detected in all of the eight carcinomas in which COX-2 protein was detected, suggesting that expression of PPARgamma and COX-2 was in a reciprocal relationship. Furthermore, in cultured ovarian carcinoma cells, Western blot revealed that PPARgamma and COX-2 expression was regulated conversely as a result of stimulation by 15-deoxy-Delta(12, 14) PGJ(2) (15-PGJ(2)), a PPARgamma activator. In addition, 15d-PGJ(2) suppressed tumour necrosis factor-alpha-induced-COX-2 expression, confirming the reciprocal correlation between COX-2 and PPARgamma. From these results, it was suggested that PPARgamma activation might suppress COX-2 expression via the nuclear factor-kappaB pathway in the ovarian carcinoma cells and that low expression of PPARgamma and high expression of COX-2 might be involved in carcinogenesis and progression of ovarian tumours.     

非小细胞肺癌患者肺组织环氧合酶-2表达的变化及其临床意义

目的探讨非小细胞肺癌(NSCLC)患者肺组织中环氧合酶-2(COX-2)的表达和血浆PGE 2含量的变化及其与临床病理参数之间的关系.方法采用逆转录聚合酶联反应(RT-PCR)和放射免疫法,分别检测38例NSCLC患者和正常健康者肺组织中COX-2 mRNA的表达以及血浆PGE 2含量的变化.结果 38例肺癌患者肺癌组织和正常肺组织中分别有34例和3例检测到COX-2 mRNA表达,阳性表达率分别为13.16%和89.47%;半定量分析肺癌组织中的COX-2 mRNA平均吸光度显著高于正常肺组织(P＜0.001).肺癌患者和正常健康者血浆中PGE 2浓度分别为(42.33±6.13)μg/L和(5.87±1.40)μg/L,二者比较,差异亦具有显著性(P＜0.01).直线相关分析肺癌组织中COX-2 mRNA的表达和PGE 2浓度的变化呈正相关(r=0.552,P＜0.05).COX-2 mRNA表达在不同性别、年龄、组织分型、TNM分期和分化程度之间差异分别均无显著性(P＞0.05).结论 NSCLC患者肺癌级织中COX-2 mRNA和PGE 2表达增强,可能在NSCLC的发生和发展中具有重要作用.     

Down-regulation of CacyBP is associated with poor prognosis and the effects on COX-2 expression in breast cancer

Calcyclin-binding protein (CacyBP) is a tumor suppressor in gastric and renal cell carcinoma, but an oncogene in pancreatic cancer. However, the function of CacyBP in breast cancer has not been well elucidated. In this study, we explored the clinical relevance of CacyBP and investigated the relationship between CacyBP and COX-2 in breast cancer. Immunohistochemical analysis in 172 cases of breast tissues showed that the positive rate of CacyBP protein expression in normal breast tissues (NBT) (89.3%) was higher than that in invasive ductal carcinoma (IDC) (56.1%) (P<0.05). RT-PCR and Western blot analysis showed that CacyBP mRNA and protein expression were significantly lower in tumor tissues as compared to those in the corresponding non-tumorous tissues (P<0.05). The expression trend of COX-2 was opposite with CacyBP in breast carcinogenesis. Moreover, the CacyBP expression was significantly negatively associated with the COX expression in the 132 breast cancer samples (correlation coefficient = 0.505, P<0.001). The clinicopathological data analysis in 132 breast cancer samples showed that CacyBP expression was positively correlated with well differentiated samples (P=0.021), low pathologic TNM stage (P=0.009), and no lymphatic metastasis (P=0.027) of patients with breast cancer. Furthermore, reduced CacyBP expression was associated with poor prognosis. Knockdown of CacyBP gene using siRNA enhanced the proliferation and invasion ability of breast cancer cells, which was dependent on COX-2 expression. In conclusion, CacyBP regulation of COX-2 expression may play an important role in human breast carcinogenesis. Restoration of CacyBP gene is a potential therapeutic target of breast cancer.     

COX-2与突变型p53蛋白在乳腺癌组织中的表达及临床意义

目的研究COX-2蛋白和突变型p53蛋白在乳腺癌组织中的表达情况,探讨两者在乳腺癌发生、发展中的作用、相互关系及与临床病理学特征之间的关系。方法运用链霉菌抗生物素蛋白-过氧化物酶免疫组织化学方法（S-P）对99例乳腺癌组织中COX-2蛋白和突变型p53蛋白的表达进行检测。结果99例乳腺癌中COX-2蛋白表达阳性率为69.7%（69/99）,COX-2蛋白表达与患者年龄、原发肿瘤大小、腋淋巴结转移状况、TNM临床分期、激素受体状况均无显著相关（P〉0.05）,而与C-erbB-2蛋白表达呈显著相关（P〈0.05）。99例乳腺癌中突变型p53蛋白表达阳性率为48.5%（48/99）,突变型p53蛋白表达与患者年龄、TNM临床分期、激素受体状况无显著相关（P〉0.05）,而与原发肿瘤大小、腋淋巴结转移状况、C-erbB-2蛋白表达呈显著相关（P〈0.05）。COX-2蛋白表达与突变型p53蛋白在乳腺癌组织中的表达呈正相关（r=0.288,P〈0.05）,两者共表达与肿瘤大小、腋淋巴结转移状况呈显著相关（P〈0.05）。结论COX-2蛋白和突变型p53蛋白在乳腺癌组织中高表达,COX-2蛋白与突变型p53蛋白在乳腺癌组织中的表达呈正相关,两者在乳腺癌的发生、发展中可能起协同作用,是反映乳腺癌生物学行为的重要指标。     

胃癌组织COX-2、VEGF-C表达与淋巴结转移及预后关系的研究

目的:探讨环氧化酶-2(cyclooxygenase-2,COX-2)、血管内皮生长因子C(VEGF-C)在胃癌组织中的表达及其与淋巴管生成、淋巴结转移及预后的关系.方法:采用免疫组织化学SABC法检测 51例胃癌及相应的癌旁组织COX-2、VEGF-C及受体VEGFR-3表达,计数肿瘤内淋巴管密度(LVD),并结合临床病理特征和随访资料进行分析.结果:胃癌组织COX-2、VEGF-C表达阳性率分别为62.8%(32/51),60.7%(31/51).COX-2表达与VEGF-C(r=O.74,P＜0.05)、临床分期(r=0.34,P＜0.05)、淋巴管密度(r=0.69,P＜0.01)和淋巴结转移(r=0.57,P＜0.01)呈正相关,与病理分化呈负相关(r=-0.58,P＜0.01).VEGF-C表达与淋巴管密度(r=0.45,P＜0.01)、淋巴结转移呈正相关(r=0.46,P＜0.05).随访5年,胃癌组织COX-2表达与生存率呈负相关,COX-2表达阴性组5年生存率(36.8%)显著高于COX-2表达阳性组(15.6%)(P＜0.05).结论:在胃癌组织中,COX-2、VEGF-C高表达,COX-2与VEGF-C、淋巴管密度、淋巴结转移呈正相关.推测COX-2通过诱导VEGF-C表达参与淋巴结转移.胃癌组织COX-2检测可能对推测预后具有重要意义.     

环加氧酶-2 通过调控EMT促进乳腺癌MDA-MB-231 细胞的迁移和侵袭

[摘要] 目的：探讨环加氧酶-2（COX-2）在乳腺癌转移中的作用及其可能的机制。方法：收集从2015 年10 月至2018 年4 月在云南省肿瘤医院接受乳腺切除术的患者中获得的原发乳腺癌组织和脑转移乳腺癌组织临床病理样本共45 例,其中原发30 例、脑转移15 例。采用qPCR检测COX-2 在原位乳腺癌和脑转移乳腺癌组织中的表达。将COX-2 过表达重组病毒（LV6-COX2）或敲减COX-2 重组病毒（LV3-COX2 shRNA1、LV3-COX2 shRNA2）感染人乳腺癌MDA-MB-231 细胞并获得稳转细胞株后,CCK-8法检测COX-2 表达对MDA-MB-231 细胞增殖的影响,划痕实验和Transwell 法检测对MDA-MB-231 细胞迁移和侵袭的影响。qPCR和WB实验分析各组细胞中COX-2 mRNA和蛋白的表达水平,qPCR检测COX-2 表达对MDA-MB-231 细胞内EMT相关基因表达的影响。结果：COX-2 表达水平在脑转移乳腺癌患者组织中显著高于原位乳腺癌组织（P<0.01）;并且与乳腺癌患者肿瘤TMN分期有关。成功构建稳定过表达/敲减COX-2 的MDA-MB-231 细胞株。过表达COX-2 促进MDA-MB-231 细胞的迁移和侵袭（均P<0.01）,同时显著提高MMP2、MMP1、N-cadherin 和vimentin 的表达（均P<0.01）,但对细胞增殖无明显影响;而沉默COX-2 则有相反的作用,且可促进细胞增殖（P<0.05）。结论：COX-2 在脑转移乳腺癌组织中高表达,其可能通过调控EMT过程促进乳腺癌MDA-MB-231 细胞的迁移和侵袭。     

15-Deoxy-Delta12,14-prostaglandin J2 induces COX-2 expression through Akt-driven AP-1 activation in human breast cancer cells: a potential role of ROS

Recent studies suggest that inflammation is causally linked to carcinogenesis. Cyclooxygenase-2 (COX-2), a rate-limiting enzyme in the biosynthesis of prostaglandins, is inappropriately expressed in various cancers and hence recognized as one of the hallmarks of chronic inflammation-associated malignancies. However, the mechanistic role of COX-2 as a link between inflammation and cancer remains undefined. Here, we report that 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), one of the final products of COX-mediated arachidonic acid metabolism, upregulates the expression of COX-2 in the human breast cancer MCF-7 cell line. 15d-PGJ(2)-induced COX-2 expression was mediated by activation of Akt and subsequently activator protein-1 (AP-1). Furthermore, 15d-PGJ(2) formed reactive oxygen species, which led to increased phosphorylation of Akt, DNA binding of AP-1 and expression of COX-2. In contrast to 15d-PGJ(2), 9,10-dihydro-15d-PGJ(2) did not elicit any of effects induced by 15d-PGJ(2) in this study, suggesting that an electrophilic carbon center present in 15d-PGJ(2) is critical for COX-2 expression as well activation of upstream signal transduction induced by this cyclopentenone prostaglandin. Taken together, these observations suggest that 15d-PGJ(2) produced by COX-2 overexpression may function as a positive regulator of COX-2 in human breast cancer MCF-7 cells.     

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。      

15-脱氧前列腺素J2对胃癌生长影响及机制的体内研究

目的 探讨过氧化物酶体增殖物激活受体gamma(PPARγ)的天然配体15-脱氧前列腺素J2(15d-PGJ2 )在体内对人胃癌细胞生长的作用以及对survivin、p27、Skp2和CD44表达的影响。方法 将人胃癌MGC803细胞,注射在12只裸鼠右上肢肩背部皮下。待长出肉眼可见的肿瘤时,将12只裸鼠随机分为实验组和对照组,每组6只。实验组静脉给予15d-PGJ2,对照组静脉给予PBS。应用反转录聚合酶链反应（RT-PCR）和Western blot方法分别检测皮下移植瘤组织中survivin、Skp2、p27的 mRNA及蛋白的表达。应用免疫组织化学法检测瘤组织CD44的表达,并对阳性细胞进行计数。结果 (1)给予15d-PGJ2 16次后,实验组裸鼠移植瘤平均体积（573.86±242.90）mm³明显小于对照组裸鼠移植瘤平均体积(1206.46±272.22) mm3（P＜0.05）。(2)实验组与对照组相比,瘤组织survivin 和Skp2的mRNA及蛋白质低表达而p27 mRNA及蛋白质高表达。(3) CD44阳性细胞率,实验组 (51.20±12.45)%明显少于对照组 (85.45±15.45)% （P＜0.05）。结论 15d-PGJ2能抑制人胃癌细胞的生长,调节survivin 、p27、Skp2和CD44的表达,提示15d-PGJ2有可能对治疗胃癌有效。