卵巢上皮癌患者血浆LPA水平检测的临床意义

目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)在卵巢上皮癌发生和发展中的作用及临床意义。方法用生物化学法检测24例卵巢上皮癌、18例卵巢良性上皮肿瘤患者和10例健康妇女的血浆LPA水平。结果卵巢上皮癌患者血浆LPA水平明显高于卵巢良性上皮肿瘤患者及健康妇女(P<0.001),后两者血浆LPA水平差异无统计学意义;卵巢上皮癌患者血浆LPA的水平Ⅲ~Ⅳ期高于Ⅰ~Ⅱ期,但是差异无统计学意义(P>0.05)。结论LPA在卵巢上皮癌患者中呈高表达,具有潜在的早期诊断价值。     

卵巢上皮性肿瘤中溶血磷脂酸受体表达及其生物学意义

目的：探讨3种溶血磷脂酸（lysophosphatidic acid，LPA）受体（LPA1，LPA2及LPA3）mRNA和蛋白在人卵巢肿瘤组织中的表达状况及意义。方法：采用半定量RT—PCR和Western blot检测LPA1、LPA2和LPA3mRNA以及LPA2和LPA3蛋白在人卵巢上皮性肿瘤组织中的表达水平。结果：RT-PCR结果显示，LPA1mRNA在正常卵巢组织、良性卵巢肿瘤及上皮性卵巢癌中表达阳性率分别为100％、100％、95．7％，差异均无显著性（P〉0．05）；上皮性卵巢癌组织LPA2mRNA、LPA3mRNA表达阳性率（95．7％；91．3％）显著高于良性卵巢肿瘤（21．3％；21．3％）及正常卵巢组织（15．4％；15．4％）（P〈0．01）；良性卵巢肿瘤与正常卵巢组织LPA2mRNA、LPA3mRNA表达阳性率差异无显著性（P〉0．05）；上皮性卵巢癌组织中LPA1mRNA表达率和表达水平高于LPA3mRNA（P〈0．05）。Western blot结果显示，上度性卵巢癌组织中LPA2和LPA3蛋白表达阳性率（95．7％；95．7％）均高于良性卵巢肿瘤（50％；42．9％）和正常卵巢组织表达阳性率（38．5％；38．5％）（P〈0．01）；良性卵巢肿瘤与正常卵巢组织中LPA2和LPA3蛋白表达阳性率及表达水平相比差异无显著性（P〉0．05）。病理资料的相关分析显示，上皮性卵巢癌组织中LPA受体mRNA和蛋自表达与临床病理特征无相关性。结论：LPA1、LPA2和LPA3在卵巢上皮性肿瘤中普遍表达，LPA受体可能参与了卵巢癌的发生发展过程。     

卵巢上皮癌患者血浆LPA和IL-6、IL-8水平的检测分析

目的探讨溶血磷脂酸(LPA)及IL-6、IL-8在卵巢上皮癌临床诊断、治疗中的价值。方法采用生化测定法和酶联免疫法,检测24例卵巢上皮癌、18例卵巢良性上皮肿瘤患者和10例健康妇女的血浆LPA和IL-6、IL-8水平。结果卵巢上皮癌组血浆LPA和IL-6、IL-8水平显著高于卵巢良性上皮肿瘤组、正常组的水平(P均<0.001),卵巢良性上皮肿瘤组与正常组比较,无显著性差异。Ⅲ、Ⅳ期患者的血浆LPA水平高于Ⅰ、Ⅱ期患者,但无显著性差异(P=0.064);Ⅲ、Ⅳ期患者的血浆IL-6和IL-8水平均明显高于Ⅰ、Ⅱ期患者(P<0.001,P<0.05)。卵巢上皮癌患者血浆LPA与IL-6,LPA与IL-8均呈正相关。结论LPA和IL-6、IL-8在卵巢上皮癌患者中呈高表达,在卵巢上皮癌的诊断、治疗中具有重要价值,且提示LPA可能通过上调卵巢上皮癌细胞IL-6和IL-8的表达,促进卵巢上皮癌进一步发展。     

溶血磷脂酸及其受体和IL-6 IL-8在乳腺癌进展中的表达变化与意义

  目的  探讨溶血磷脂酸(lysophosphatidic acid, LPA)及其受体和IL-6与IL-8在乳腺癌进展中的表达及临床意义。  方法  采用半定量RT-PCR方法检测乳腺肿瘤组织和瘤旁组织中LPA受体的表达水平。采用LPA生化测定法和酶联免疫吸附(ELISA)法分别检测乳腺肿瘤患者和健康妇女的血浆LPA、IL-6和IL-8水平。  结果  术后复发转移乳腺癌患者血浆LPA、IL-6、IL-8的表达水平均显著高于局限期乳腺癌和良性乳腺肿瘤患者及健康妇女(P均 < 0.05);LPA1在乳腺癌组织中的表达水平显著高于良性乳腺肿瘤组织和正常乳腺组织(P < 0.05);乳腺癌患者血浆LPA水平与IL-6和IL-8水平均呈正相关(P均 < 0.01)。  结论  LPA对乳腺癌患者内源性IL-6和IL-8的表达可能具有上调作用, 检测LPA、IL-6和IL-8的表达水平对乳腺癌的转移可能有一定的预测作用, 尤其是骨转移。      

卵巢及卵巢肿瘤组织FAS和FASL表达的初步研究

目的:探讨FAS、FASL与上皮性卵巢癌的关系.方法:采用免疫组化法检测10例正常卵巢组织、30例良性卵巢上皮肿瘤、32例交界性卵巢上皮性肿瘤和52例卵巢上皮癌组织中FAS、FASL的表达情况,分析两者在卵巢上皮性肿瘤发病机制中的作用.结果:卵巢上皮癌组织中FAS的表达明显低于交界性卵巢上皮肿瘤、良性卵巢上皮肿瘤和正常卵巢组织,差异有统计学意义,H=20.784,P＜0.001;而正常卵巢组织、良性卵巢上皮肿瘤、交界性卵巢上皮肿瘤之间Fas表达差异无统计学意义,P＞0.05.卵巢上皮癌组织中FASL的表达明显高于良性卵巢上皮肿瘤和正常卵巢组织,差异有统计学意义,P＜0.05;而交界性卵巢上皮肿瘤、良性卵巢上皮肿瘤和正常卵巢组织之间的FASL表达差异无统计学意义,P＞0.05.结论:FAS在卵巢上皮癌组织中表达下调;FASL在卵巢上皮癌组织中表达上调.FAS、FASL在卵巢上皮癌的发生发展过程中可能通过共同的机制参与卵巢上皮癌的发生.     

溶血磷脂酸受体-1在乳腺癌组织中的表达

目的探讨溶血磷脂酸(lysophosphatidicacid,LPA)受体的三种亚型(LPA1、LPA2和LPA3)在乳腺癌组织中的表达水平及与乳腺癌发生发展的关系。方法采用半定量RT-PCR方法检测乳腺癌组织(35例)、癌旁组织(14例)、乳腺纤维瘤组织(15例)和正常乳腺组织(8例)中的LPA受体的三种亚型mRNA的表达水平,并对乳腺癌组织中这些受体的表达水平与病理类型、肿瘤大小和临床分期等的关系进行分析。结果LPA1在乳腺癌组织中的表达水平显著高于乳腺纤维瘤组织(P<0.05)和正常乳腺组织(P<0.05);最大直径≥2cm乳腺癌组显著高于直径<2cm组(P<0.05);浸润性导管癌也显著高于其它浸润性乳腺癌(P<0.05)。LPA2与LPA3在乳腺癌、癌旁、乳腺纤维瘤和正常乳腺四种组织中的表达水平均无统计学差异。结论LPA1在乳腺癌组织中表达增高,可能在乳腺癌发生与发展的过程中有重要的作用。     

人表皮生长因子2及拓扑异构酶Ⅱα在卵巢上皮肿瘤的表达及相关性检测

目的探讨人类表皮生长因子2（HER-2／NEU）及拓扑异构酶Ⅱα（TopoⅡα）在卵巢上皮肿瘤的表达及二者的相关性的检测。方法采用免疫组化S-P法检测53例卵巢上皮癌，19例良性卵巢上皮肿瘤及15例正常卵巢组织的HER-2／NEU及TopoⅡα的蛋白水平的表达。结果免疫组化结果显示：卵巢上皮癌组织中HER-2，TopoⅡα的阳性表达率分别是47．2％，66％。明显高于良性上皮肿瘤及正常卵巢组织中的表达。对二者相关性进行检测，二者表达正相关，（rs=0．358 P〈10．05）。结论HER-2，TopoⅡα在卵巢恶性上皮肿瘤的表达显著高于在卵巢良性上皮肿瘤和正常卵巢组织的表达，且两者的表达相关。提示两基因表达的上调可能在卵巢恶性上皮肿瘤的发生及发展中起一定的作用。     

血浆溶血磷脂酸及CA125水平与子宫内膜癌临床病理特征的关系

目的:探讨血浆溶血磷脂酸（LPA）及CA125在子宫内膜癌的表达及与临床病理特征之间的关系。方法:利用化学比色法检测65例子宫内膜癌患者和40例子宫内膜非典型增生患者及40例健康献血员（对照组）血浆溶血磷脂酸含量,同时采用电化学发光法检测血浆CA125含量并作对比分析。结果:子宫内膜癌组患者血浆LPA水平为（5.87±2.02）μmol/L,明显高于对照组（2.31±0.45）μmol/L（P<0.01）,子宫内膜非典型增生组与对照组比较,差异无统计学意义（P>0.05）。LPA和CA125对子宫内膜癌诊断的灵敏度分别为69.2%、63.1%,特异度为60.0%、53.8%;LPA对子宫内膜癌早期（0+Ⅰ期）诊断的阳性率明显高于晚期（Ⅱ～Ⅳ期）（P<0.05）。与肿瘤大小无明显的相关性（P>0.05）。其阳性率随病理分级逐渐降低并有显著性差异（P均＜0.05）,且随肿瘤浸润肌层深度及淋巴结转移程度增加亦逐渐降低（P均＜0.05）。而CA125阳性率随着病理分级逐渐升高,并有显著性差异（P均＜0.01）,在深肌层浸润及子宫外转移病例中明显升高（P均＜0.01）。结论:血浆LPA检测对子宫内膜癌的早期诊断及鉴别诊断具有一定的临床应用价值,明显优于CA125,LPA可作为判断子宫内膜癌预后的重要指标。     

胰腺癌患者血浆溶血磷脂酸的临床意义

目的:检测胰腺癌患者血浆溶血磷脂酸(lysphosphaticlic acid,LPA)的水平,评价LPA在胰腺癌诊治中的临床意义。方法:采用定磷法检测2006年6月至2010年10月南京第一医院收治的胰腺癌患者50例、胰腺良性疾病患者32例及健康志愿者36人的血浆LPA水平,同时测定血清CA19-9、AFP和CEA的水平;免疫组化法检测胰腺癌组织及癌旁胰腺组织LPA2受体的表达。分析血浆LPA水平与胰腺癌临床病理特征的关系。结果:胰腺癌患者血浆LPA水平明显高于胰腺良性疾病患者和健康志愿者[(4.10±2.03)vs(3.28±1.26)、(2.27±1.02)μmol/L,P<0.05],胰腺癌患者血浆LPA水平和血清CA19-9水平密切相关(r=0.9070,P<0.01)。胰腺癌组织LPA2受体表达阳性率显著高于癌旁正常胰腺组织(88%vs 4%,P<0.05)。血浆LPA水平的升高与胰腺癌浸润和淋巴结转移等相关。结论:血浆LPA检测为胰腺癌诊断和预后判断增加了一项潜在的评价指标。     

反义寡聚脱氧核苷酸对卵巢癌细胞系LPA受体抑制与对MMP-2和MMP-9的调控

目的:研究溶血磷脂酸受体(LPA1、LPA2、LPA3)的反义寡核苷酸(ASODN)对卵巢癌细胞的抑制作用;探讨LPA对表达LPA受体ASODN的卵巢癌细胞株MMP2和MMP9表达及活性的影响.方法:将LPA1、LPA2和LPA3ASODN转入卵巢癌细胞系SKOV3和3AO,用RT-PCR和蛋白质印迹法鉴定转染细胞株的表达水平.通过检测细胞株生长、增殖变化观察转染细胞株的生物学特性,明胶酶谱检测分析LPA对表达LPA受体的ASODN的卵巢癌细胞株MMP-2、MMP-9表达及活性的影响.结果:LPA1、LPA2及LPA3 ASODN均可有效的抑制LPA1基因、LPA2及LPA3蛋白质的表达.转染前后细胞株生长情况差异无统计学意义.明胶酶谱条带的灰度值分析显示,LPA诱导表达LPA2、LPA3 ASODN的卵巢癌细胞株MMP-2、MMP-9活性降低,与对照组及正义组相比,差异有统计学意义,P<0.05.转染LPA1 ASODN的卵巢癌细胞株MMP-2、MMP-9活性,与对照组及正义组相比,差异无统计学意义,P>0.05.结论:ASODN抑制LPA受体基因及蛋白质表达.在LPA诱导的MMPs活化过程中,LPA2及LPA3更有效,提示卵巢上皮性癌中LPA2和LPA3在LPA诱导的MMPs侵袭、转移过程中发挥主要作用.     

Lack of significant differences in the corrected activity of lysophospholipase D, producer of phospholipid mediator lysophosphatidic acid, in incubated serum from women with and without ovarian tumors

BACKGROUND: Several studies have shown that lysophosphatidic acid (LPA), a phospholipidic chemical mediator, is relevant to the pathogenesis of ovarian carcinoma. Higher plasma levels of LPA have been reported in patients with ovarian carcinoma than in healthy patients, and LPA is known to activate ovarian carcinoma cells. To determine the reason for the increased plasma LPA levels in ovarian carcinoma patients, we compared the activities of serum lysophospholipase D, a novel LPA-producing metallo-enzyme, in healthy volunteers, patients with benign ovarian tumor, and patients with ovarian carcinoma. METHODS: Lysophospholipase D activity was assessed by measuring the percentage conversion of [14C]palmitoyl-lysophosphatidylcholine (LPC) added to human serum. The apparent enzyme activities were corrected based on the serum levels of palmitoyl-LPC determined by gas-liquid chromatography after its purification and conversion to fatty acid methyl esters. RESULTS: The apparent activity of lysophospholipase D in serum preparations from four patients with ovarian carcinoma at Stage IV was significantly higher than those from five healthy subjects, five patients with benign ovarian tumors, and fourteen patients with ovarian carcinoma at Stages I (n = 5), II (n = 4), and III (n = 5). The serum levels of LPC, an endogenous substrate of lysophospholipase D, in ovarian carcinoma patients were less than those in patients with benign ovarian tumors. There were no significant differences in the corrected lysophospholipase D activity for the LPC levels in healthy women, patients with benign ovarian tumors, and patients with ovarian carcinoma at various stages. CONCLUSIONS: The current results suggest that lysophospholipase D is not associated with the elevated plasma levels of LPA in ovarian carcinoma patients previously reported, although only a limited number of patients were analyzed.     

Expression of Vascular Endothelial Growth Factor-C and Vascular Endothelial Growth Factor Receptor-3 in Ovarian Epithelial Tumors

Objective： To explore the role of vascular endothelial growth factor-C （VEGF-C） in the process of angiogenesis, lymphangiogenesis and lymphatic metastasis in epithelial ovarian tumors. Methods： In situ hybridization and immunohistochemical staining for VEGF-C were performed in 30 epithelial ovarian carcinomas, 9 borderline tumors and 26 benign tumors. Endothelial cells were immunostained with anti-VEGFR-3 pAb and anti-CD31 mAb, and VEGFR-3 positive vessels and microvessel density （MVD） were assessed by image analysis. Results： VEGF-C mRNA and protein expression were detected in cytoplasm of carcinoma cells. VEGF-C mRNA and protein expression in ovarian epithelial carcinomas were significantly higher than those in borderline tumors and benign tumors （P〈0.05 or P〈0.01）. In ovarian epithelial carcinomas, VEGF-C protein expression, VEGFR-3 positive vessels and MVD were significantly higher in the cases of clinical stage Ⅲ-Ⅳ and with lymph node metastasis than those of clinical stage Ⅰ-Ⅱ and without lymph node metastasis respectively （P〈0.05 or P〈0.01）. VEGFR-3 positive vessels and MVD were significantly higher in VEGF-C protein positive tumors than negative tumors （P〈0.05）. VEGFR-3 positive vessels was significantly correlated with MVD（P〈0.01）. Conclusion： VEGF-C might play a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in epithelial ovarian tumors, and VBEGF-C could be used as a biologic marker of metastasis in ovarian epithelial tumors.     

微管相关蛋白LC3和自噬基因Beclin1在上皮性卵巢癌中的表达及其意义

背景与目的:有研究表明自噬活性的改变与肿瘤的发生、发展有关。诱导自噬性细胞死亡已经成为杀死肿瘤细胞的新策略。本研究检测微管相关蛋白LC3和自噬基因Beclin1在不同卵巢肿瘤组织中的表达情况,探讨其与上皮性卵巢癌发生、发展的相关性及意义。方法:用免疫组化法检测25例良性卵巢肿瘤、25例交界性卵巢肿瘤及75例上皮性卵巢癌组织的LC3和Beclin1表达,并结合临床病理因素进行分析。结果:良性肿瘤组LC3和Beclin1的阳性率(100%、100%)和交界性卵巢肿瘤组LC3和Beclin1的阳性率(96%、84%)都明显高于上皮性卵巢癌组(57%、57%),且差异具有统计学意义(P均<0.001)。LC3在上皮性卵巢癌中的表达与FIGO分期和肿瘤分化程度相关(P=0.017;0.001)。Beclin1表达与上皮性卵巢癌患者FIGO分期相关(P=0.04)。LC3和Beclin1在卵巢上皮癌中的表达无相关性(P=0.875)。结论:LC3和Beclin1在上皮性卵巢癌中的表达下调,自噬活性的改变可能与上皮性卵巢癌的发生、发展相关。     

三种血清肿瘤标志物联合检测对卵巢癌诊断的临床应用价值

目的:探讨溶血磷脂酸(lysophosphatidic acid,LPA)、恶性肿瘤特异性生长因子（tumor specific growth factor,TSGF）和糖类抗原125（cancer antigen 125,CA125）联合检测对卵巢癌诊断的临床应用价值。方法:对108例卵巢癌和43例卵巢良性肿瘤和50例健康女性体检者的血清中LPA、TSGF和CA125进行测定,分析肿瘤标志物联合检测对卵巢癌的诊断价值。结果:卵巢癌组LPA、TSGF和CA125水平均显著高于良性卵巢肿瘤组和正常对照组(P均<0.01);LPA、TSGF和CA125联合检测的灵敏度和特异度高于单项和两两联合检测。结论:肿瘤标志物联合检测可提高对卵巢癌诊断的准确性,对卵巢癌的鉴别诊断提供可靠的实验依据。     

Clinical significance of Skp2 expression, alone and combined with Jab1 and p27 in epithelial ovarian tumors

We have previously demonstrated the inverse correlation of Jab1 and p27 proteins, as well as prognostic significance in epithelial ovarian carcinomas. In order to investigate Skp2 protein and its correlation with Jab1, p27, and clinical outcome, we evaluated Skp2 expression in a group of epithelial ovarian tumors. Immunohistochemical analysis was performed on 80 cases of ovarian tumors (33 benign and 47 malignant), and 26 of the 80 cases were evaluated by Western blot analysis. Immunofluorescence was carried out in the human ovarian adenocarcinoma cell line OVCAR-3. Skp2 expression was detected in 53.2% of malignant tumors and 18.2% of benign tumors. The positive ratio of Skp2 expression was increased from benign to malignant ovarian tumors (p=0.002). A negative correlation between Skp2 and p27 was found in benign and malignant ovarian tumors (p=0.006 and p<0.0001, respectively). Skp2 expression was significantly associated with high tumor grade (p=0.001), lymph node metastasis (p=0.01), and residual disease (p=0.012). Kaplan-Meier survival analysis showed that Skp2 expression was significantly associated with poor prognosis (p=0.013), and patients with Skp2(+)/Jab1(+)p27(-) expression had the worst prognosis among all phenotypes of Skp2/Jab1/p27 expression (p=0.0007). Our results suggest that Skp2 expression was significantly associated with malignancy, and the Skp2 protein level may be a valuable prognostic factor for epithelial ovarian carcinomas. Furthermore, the combined evaluation of Skp2/Jab1/p27 proteins provides important prognostic information on patients with epithelial ovarian carcinoma.     

p27和CyclinE在卵巢上皮性肿瘤中的表达及其临床意义

目的 :研究p2 7、CyclinE与卵巢上皮性肿瘤发生及发展的关系。方法 :采用SP免疫组织化学方法检测了 4 0例卵巢恶性上皮肿瘤、6例交界性肿瘤及 18例良性肿瘤中p2 7、CyclinE蛋白的表达情况。结果 :p2 7和CyclinE在良性肿瘤与交界性肿瘤中表达差异无显著意义。p2 7在恶性肿瘤中的表达率 (35 0 % )显著低于交界性肿瘤和良性肿瘤表达率 (77 78%、4 6 ) ,P <0 0 1;CyclinE在卵巢上皮恶性肿瘤中的表达率显著高于良性肿瘤及交界性肿瘤 ,P <0 0 1。p2 7和CyclinE蛋白表达与卵巢癌的临床分期、病理分级、淋巴结转移有关。p2 7阴性 CyclinE阳性者 ,其生物学行为极差。结论 :p2 7、CyclinE与卵巢癌的发生及发展密切相关 ,其异常表达提示卵巢肿瘤预后差     

间皮素mRNA及蛋白在卵巢癌中的表达及分析

目的 研究间皮素(MESO)在卵巢癌中的表达及意义.方法 用逆转录聚合酶链反应(RT-PCR)技术和免疫组化方法分别检测卵巢肿瘤和正常卵巢组织中MESO mRNA及其蛋白水平.结果MESO mRNA和蛋白在上皮性卵巢癌(1.4005 ±0.4646,2.7857±2.2712)和交界性卵巢肿瘤(1.0650 ±0.3100,2.9167 ±2.391)中的表达水平高于良性卵巢肿瘤(0.6463±0.2419,1.2500 ±1.6125)和正常卵巢组织(0.6439 ±0.2729,0.9167 ±1.2401),差异有统计学意义(P＜0.05);MESOmRNA和蛋白在浆液性卵巢癌(1.5255 ±0.4151,3.3036 ±2.6141)和子宫内膜样癌(1.5250 ±0.5419,3.0000 ±2.3094)中的表达水平高于黏液样癌(1.0675±0.3149,1.0556 ±1.9242),差异有统计学意义(P＜0.05);临床Ⅲ、Ⅳ期MESO表达水平(1.5100 ±0.4142,3.6087 ±3.3959)高于Ⅰ、Ⅱ期(1.1190 ±0.4909,1.7895 ±2.6320),差异有统计学意义(P＜0.05).MESO表达水平与病理分级有关(P＜0.05),而与患者年龄和血清CA125水平无关(P＞0.05).结论 MESO mRNA及蛋白在卵巢癌和交界性肿瘤组表达增高,MESO可能参与卵巢癌的黏附转移.