Alteration and clinical relevance of PTEN expression and its correlation with survivin expression in epithelial ovarian tumors

The tumor suppressor PTEN, phosphatase and tensin homolog on chromosome 10, plays an essential role in regulating signaling pathways involved in cell growth and apoptosis and is inactivated in a wide variety of tumors. Survivin, a member of the inhibitor of apoptosis protein family (IAP), is associated with cell proliferation, and overexpressed in common human tumors. Both PTEN and survivin proteins can regulate cell cycle and apoptosis, but their biological effects are adverse. We have previously investigated the role of survivin expression in epithelial ovarian tumors. In this study, we evaluated the alteration and clinical relevance of PTEN expression and further assessed its correlation with survivin expression in epithelial ovarian tumors. Immunohistochemical analysis was performed in 103 cases of ovarian tumors, and 26 of the 103 cases were evaluated by Western blot analysis. PTEN expression was reduced from benign to malignant ovarian tumors (p=0.0003), and an inverse correlation between PTEN and survivin was found in benign, borderline, and malignant tumors (p=0.004, p=0.015 and p=0.0005, respectively). PTEN expression was significantly associated with tumor grade (p=0.001), histological subtype (p=0.037), ascites (p=0.038), and residual disease (p=0.0006). Kaplan-Meier survival analysis showed that the loss of PTEN expression was significantly associated with poor overall survival (p=0.021), and patients with PTEN(-)/survivin(+) expression had the worst prognosis among all phenotypes of PTEN/survivin expression (p=0.039). Our results suggest that the altered PTEN expression and its inverse correlation with survivin may be involved in the development and progression of ovarian tumors, and the combined detection of PTEN and survivin proteins might be more valuable in the evaluation of malignancy and prognosis in epithelial ovarian tumors.     

p21-激活激酶1基因在卵巢上皮性肿瘤中的过度表达及其机制和意义

目的探讨p21-激活激酶1（PAK1）基因在卵巢上皮性肿瘤中的过度表达及其机制和意义。方法运用免疫组化、荧光原位杂交和末端脱氧核苷酸转移酶介导缺口末端标记方法,结合组织芯片技术,检测PAK1基因在30例卵巢腺瘤、20例卵巢交界性肿瘤和80例卵巢癌中的表达、扩增及其细胞凋亡情况。结果在免疫组化有效检测的病例中,7例（25．9％）卵巢良性腺瘤、7例（36．8％）交界性肿瘤和53例（68．8％）卵巢癌出现PAK1蛋白的过度表达,而且PAK1蛋白过表达与卵巢肿瘤的细胞凋亡指数呈负相关（P=0．002）。此外,87．1％（27／31）的低分化卵巢癌（Silverberg G3级）出现PAK1蛋白的过度表达,其过表达率显著高于G1～G2级的卵巢癌（26／46,56．5％;P=0．01）。荧光原位杂交结果显示,只有2例（4．7％）卵巢癌出现PAK1基因扩增,卵巢交界性肿瘤和良性腺瘤均未观察到PAK1基因的扩增。结论PAK1蛋白过度表达可能在卵巢上皮性肿瘤的发生发展中起重要作用,而且与卵巢癌的恶性组织学表型密切相关;在卵巢肿瘤PAK1蛋白的表达调控中,基因扩增以外的其他调节机制可能起更为关键的作用。     

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     

卵巢癌中EIF-5A2 基因表达与临床意义*

目的:探讨真核翻译起始因子5A2（EIF-5A2）基因在卵巢上皮性肿瘤中的表达及其临床意义。方法:运用免疫组化和荧光原位杂交方法,结合组织芯片技术,检测EIF-5A2 基因在50例卵巢腺瘤、50例卵巢交界性肿瘤和150 例卵巢癌中的表达,分析其与肿瘤临床病理学参数之间的相关性。结果:免疫组化检测结果,分别有6.4% 的卵巢良性腺瘤、28.3% 的交界性肿瘤和56.6% 的卵巢癌出现EIF-5A2 蛋白的过度表达。在卵巢癌中,EIF-5A2 蛋白表达与肿瘤的组织学Silverberg 氏分级和临床FIGO分期均有显著的相关性（P<0.05）,其中70.0% 的高级别（G3 级）的卵巢癌出现EIF-5A2 蛋白的过度表达,明显高于G1/G2 级的卵巢癌（49.5%）;在FIGO分期中,65.6% 的临床晚期（Ⅲ/Ⅳ期）卵巢癌呈EIF-5A2 蛋白过度表达,明显高于Ⅰ/Ⅱ期的肿瘤（38.3%）。 另外,EIF-5A2 蛋白过度表达与卵巢癌细胞增殖（Ki-67的表达水平）显著正相关（P<0.01）,大多数（72.8%）EIF-5A2 蛋白过度表达的卵巢癌中出现Ki-67蛋白高表达,而多数（66.1%）EIF-5A2 蛋白正常表达的卵巢癌则呈Ki-67蛋白低表达。荧光原位杂交结果显示,只有13.8% 的卵巢癌出现EIF-5A2 基因扩增;卵巢交界性肿瘤和良性腺瘤中均未观察到EIF-5A2 基因的扩增。结论:EIF-5A2 蛋白过度表达可能通过促进肿瘤细胞增殖的效应,在卵巢上皮性肿瘤的发生发展中起重要作用,而且与卵巢癌的恶性组织学表型和浸润转移密切相关。      

APRIL和RegⅣ在卵巢癌中的表达及意义

目的检测APRIL和RegⅣ在卵巢肿瘤和正常组织中的表达情况及其临床意义。方法采用免疫组化染色,光镜观察结果确定APRIL和RegⅣ在良性、交界性及恶性上皮性卵巢肿瘤组织中的表达,并结合临床病理资料进行分析。结果在良性、交界性及恶性上皮性卵巢肿瘤组织中皆有APRIL和RegⅣ的表达,但卵巢癌组织中的表达明显高于良性及交界性肿瘤,其表达强度与淋巴结转移、病理类型、病理分级、年龄等无关。相关分析表明APRIL和RegⅣ的表达呈正相关。结论APRIL和RegⅣ基因与癌组织的发生和进展密切相关。细胞的增殖和机体的免疫调控以及凋亡途径有着密切联系。     

PCNA及bcl-2与caspase-3在卵巢上皮性交界性肿瘤中的表达及意义

目的:探讨增殖细胞核抗原(PCNA)、bcl-2与caspase-3在卵巢上皮性交界性肿瘤中的表达及临床意义.方法:选取天津医科大学附属肿瘤医院病理科存档的卵巢上皮性交界性肿瘤40例,采用免疫组化SP法检测PCNA、bcl-2与caspase-3在肿瘤组织中的表达,并与良性、恶性肿瘤进行比较.结果:在卵巢交界性肿瘤中1)PCNA阳性率为72.5%,明显高于良性肿瘤(47.6%),低于恶性肿瘤(89.6%,P＜0.05);bcl-2与caspase-3的阳性率为60.0%和72.5%,而良性与恶性肿瘤分别为61.9%和76.2%、86.2%和44.8%,二者在恶性肿瘤中的表达与良性、交界性有显著差异(P＜0.05).2)PCNA与bcl-2的表达存在相关性,其协同表达率为52.5%,与良性(28.6%)、恶性肿瘤(75.9%),存在显著性差异(P＜0.05);PCNA、bcl-2与caspase-3的协同表达率为72.7%,良性与交界性肿瘤明显高于恶性肿瘤(P＜0.05).3)仅bcl-2与组织学类型有关,浆液性肿瘤的阳性率明显高于粘液性(P＜0.01);PCNA、caspase-3与临床病理参数无关.结论:PCNA与bcl-2任一高表达尤其协同表达,并伴caspase-3低表达是卵巢交界性肿瘤恶性潜能增加的标志,应密切随访.     

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.     

应用组织微阵列和原位杂交检测卵巢上皮性肿瘤中HIF-1α mRNA的表达

目的　探讨缺氧诱导因子 1α(HIF 1α)在卵巢上皮性肿瘤中的表达及意义。方法　采用组织微阵列 (TMA)新技术和原位杂交方法 ,回顾性研究 2 95例卵巢上皮性肿瘤中HIF 1αmRNA的表达情况 ,并以 13例正常卵巢组织作对照。结果　正常卵巢、卵巢上皮性良性肿瘤、交界性肿瘤和卵巢癌组织的HIF 1αmRNA表达阳性率分别为 0、13.2 %、4 2 .1%和 81.9%。交界性肿瘤和卵巢癌组织中的HIF 1αmRNA表达显著高于正常卵巢和良性卵巢上皮性肿瘤 (P <0 .0 0 1) ,卵巢癌显著高于交界性肿瘤 (P <0 .0 0 1)。卵巢癌组织中HIF 1αmRNA的表达与临床病理分期和病理类型无相关性 (P >0 .0 5 )。但在不同组织学分级 (即高、中、低分化 )的卵巢癌组织中 ,HIF 1α表达率分别为 6 0 .4 %、81.2 %和 96 .4 % ,与组织学分化程度呈负相关 (P <0 .0 0 1)。结论　HIF 1α在卵巢癌的发生、发展过程中可能起着重要作用。组织微阵列技术适用于大样本量组织的检测 ,是一种高效、快速、可比性强的分子生物学研究方法。     

Overexpression of the MET/HGF receptor in ovarian cancer

The MET oncogene encodes the receptor for Hepatocyte Growth Factor/Scatter Factor, a unique growth factor that induces not only proliferation of epithelial cells, but also cell motility and invasiveness. DNA level and expression of the Met/HGF receptor gene were examined with Southern- and Western-blot analyses, respectively, in human ovary, benign ovarian tumors and epithelial ovarian carcinomas. The Met/HGF receptor was detectable in the surface epithelium of normal ovary. The level of expression was unchanged in benign ovarian tumors of various origins. Fourteen out of 67 malignant carcinomas (20%) showed a 3-to 10-fold increase in expression. In 5 additional cases the Met/HGF protein was overexpressed over 50-fold. This represents a total of 28% of cases. Overexpression was not associated with MET gene amplification. Overexpressing tumors belonged to different histotypic variants, but showed a well-differentiated phenotype. Clinically, overexpression was associated with disease at any pathologic stage, but was significantly correlated with premenopausal status of patients. These data suggest that expression of the Met/HGF receptor may add a selective growth advantage to a narrow subset of differentiated ovarian cancers in premenopausal patients.     

Overexpression of follicle-stimulating hormone receptor facilitates the development of ovarian epithelial cancer

We previously showed that the expressing level of FSH receptor (FSHR) increased from ovarian epithelial inclusions (OEIs) to benign ovarian epithelial tumors (OETs) and to borderline OETs, whereas FSHR levels decreased with an increase in carcinoma grade. The aim of this study was to investigate the role of FSHR in OET development. MCV152 cells with FSHR overexpression showed an increased cellular proliferation and invasive capacity, which was associated with reduced levels of prohibitin and RII-β expression and increased levels of HER-2/neu, c-Myc, and EGFR expression. Overexpression of FSHR may be associated with an elevated level of OET cell proliferation via an enhanced activity of potential oncogenic pathways. Therefore, the findings in this study suggest that overexpression of FSHR may play a role in OET development.     

Phosphorylated 4E binding protein 1: a hallmark of cell signaling that correlates with survival in ovarian cancer

BACKGROUND: Growth factor receptors and cell signaling factors play a crucial role in human carcinomas and have been studied in ovarian tumors with varying results. Cell signaling involves multiple pathways and a myriad of factors that can be mutated or amplified. Cell signaling is driven through the mammalian target of rapamycin (mTOR) and extracellular regulated kinase (ERK) pathways and by some downstream molecules, such as 4E binding protein 1 (4EBP1), eukaryotic initiation factor 4E, and p70 ribosomal protein S6 kinase (p70S6K). The objectives of this study were to analyze the real role that these pathways play in ovarian cancer, to correlate them with clinicopathologic characteristics, and to identify the factors that transmit individual proliferation signals and are associated with pathologic grade and prognosis, regardless specific oncogenic alterations upstream. METHODS: One hundred twenty-nine ovarian epithelial tumors were studied, including 20 serous cystadenomas, 7 mucinous cystadenomas, 11 serous borderline tumors, 16 mucinous borderline tumors, 29 serous carcinomas, 16 endometrioid carcinomas, 15 clear cell carcinomas, and 15 mucinous carcinomas. Tissue microarrays were constructed, and immunohistochemistry for the receptors epidermal growth factor receptor (EGFR) and c-erb-B2 was performed and with phosphorylated antibodies for protein kinase B (AKT), 4EBP1, p70S6K, S6, and ERK. RESULTS: Among 129 ovarian neoplasms, 17.8% were positive for c-erb-B2, 9.3% were positive for EGFR, 47.3% were positive for phosphorylated AKT (p-AKT), 58.9% were positive for p-ERK, 41.1% were positive for p-4EBP1, 26.4% were positive for p70S6K, and 15.5% were positive for p-S6. Although EGFR, p-AKT, and p-ERK expression did not differ between benign, borderline, or malignant tumors, c-erb-B2, p-4EBP1, p-p70S6K, and p-S6 were expressed significantly more often in malignant tumors. Only p-4EBP1 expression demonstrated prognostic significance (P = .005), and only surgical stage and p-4EBP1 expression had statistical significance in the multivariate analysis. CONCLUSIONS: In patients with ovarian carcinoma, significant expression of p-4EBP1 was associated with high-grade tumors and a poor prognosis, regardless other oncogenic alterations upstream. This finding supports the study of this factor as a hallmark or pivotal factor in cell signaling in ovarian carcinoma that may crucial in the transmission of the proliferation cell signal and may reflect the real oncogenic role of this pathway in ovarian tumors.     

PKP4在卵巢上皮性浆液性囊腺癌组织中的表达及其临床意义

目的:检测PKP4（plakophilin 4,又称p0071）在卵巢上皮性浆液性囊腺癌组织中的表达,探讨PKP4参与肿瘤发生发展的机制及其临床意义。方法:利用免疫组化SP法检测PKP4在55例卵巢上皮性浆液性囊腺癌,12例卵巢上皮性交界性肿瘤,13例卵巢上皮性良性肿瘤,15例正常卵巢组织中表达情况,分析其与卵巢上皮性浆液性囊腺癌患者临床病理参数及预后的关系。结果:PKP4以细胞膜、细胞质着色为主,PKP4在卵巢上皮性浆液性囊腺癌组织中阳性表达率(94.55%)明显高于正常卵巢组织(26.67%)、卵巢上皮性良性肿瘤组织(46.15%)及卵巢上皮性交界性肿瘤(75.00%)(P＜0.05)。PKP4表达随FIGO分期增加而增高(P＜0.05),是影响卵巢上皮性浆液性囊腺癌患者总生存时间和预后的独立危险因素。结论:PKP4与卵巢癌的发生、发展和预后相关,有望在卵巢癌的早期诊断及预后评估方面发挥重大意义。     

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.     

自噬基因Beclin1与上皮性卵巢癌发生、发展的相关性研究

背景与目的:有研究表明自噬的抑制与肿瘤的发生有关,沉默自噬基因Beclin1可导致多种恶性肿瘤的发生.本研究探讨Beclin1基因在上皮性卵巢癌发生、发展中的作用,并分析其过表达对人卵巢癌细胞株SKOV3体外生长的影响.方法:用免疫组化检测25例正常卵巢组织、25例良性上皮性卵巢肿瘤、19例交界性上皮性卵巢肿瘤及69例上皮性卵巢癌组织的Beclin1表达;构建真核表达载体pcDNA3.1/Beclin1,脂质体法分别将质粒pcDNA3.1/Beclin1、pcDNA3.1转染人卵巢癌细胞株SKOV3,用MTT法分析外源性Beclin1过表达对SKOV3增殖的影响,并用流式细胞仪检测凋亡情况.结果:正常卵巢组织和良性卵巢肿瘤组织中Beclin1表达的积分光密度(integrated optical density,IOD)值均较高,两者之间的差异无显著性(P＞0.05);交界性卵巢肿瘤组织中Beclin1的表达降低,而在上皮性卵巢癌中Beclin1表达的IOD值最低,与正常卵巢组织比较差异有显著性(P＜0.05).ocDNA3.1/Beclin1转染SKOV3细胞后细胞生长抑制率为(68.75±5.10)%,而pcDNA3.1转染组为(10.91±4.20)%,两者之间差异有显著性(P＜0.05).pcDNA3.1/Beclin1转染后72 h SKOV3细胞的凋亡率为19.07%,高于pcDNA3.1转染组和未转染组,差异有显著性(P＜0.05).结论:Beclin1在上皮性卵巢癌中的表达下调,可能与上皮性卵巢癌的发生、发展有关;自噬基因Beclin1的过表达可抑制人卵巢癌细胞SKOV3的增殖,并诱导其凋亡.     

卵巢上皮癌组织中survivin的表达及其与Fas、FasL表达的关系

背景与目的:近期研究表明,凋亡抑制蛋白survivin在细胞凋亡和有丝分裂中起重要作用,于多种常见的恶性肿瘤有表达。本研究拟探讨survivin在卵巢上皮癌中的表达,及其与Fas、FasL表达的相互关系。方法:用免疫组织化学SP法检测84例卵巢上皮癌组织、39例卵巢良性肿瘤组织和20例正常卵巢组织中survivin、Fas、FasL的表达。结果:63.1%的卵巢上皮癌组织表达survivin基因,良性肿瘤组织表达率为30.8%,而正常卵巢组织中无表达,差异有显著性(P<0.01)。Survivin表达的高低与临床分期、组织学分级等密切相关。Fas在卵巢上皮癌组织中的表达(23.8%)显著低于卵巢良性肿瘤组织(53.8%)(P<0.01);而FasL在卵巢上皮癌组织中的表达(44.0%)则显著高于卵巢良性肿瘤组织(23.1%)(P<0.05)。survivin的表达与Fas、FasL的表达密切相关。结论:(1)Survivin基因在卵巢上皮癌组织中表达上调;(2)Fas与FasL在卵巢上皮癌中的异常表达及其与survivin基因的表达密切相关,说明它们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。     

Weapons Ovarian Epithelial Tumors May Use in Immune Escape: An Immunohistochemical Correlational Study

Investigate FasL and survivin expression in a series of primary ovarian surface epithelial tumors, correlate their expression with each other, and characterize the presence of CD3+ T-lymphocytes in studied tumors and determine whether their presence correlates with FasL or survivin expression in malignant cases. FasL and survivin expression was assessed in 54 ovarian epithelial tumors. The results were compared between different tumor types and grades. Correlation between both markers’ expression in all studied tumors was done. Either marker’s expression was compared to the mean CD3+ T-lymphocytes per HPF in the studied malignant tumors. Either FasL or survivin expression was significantly higher in malignant versus benign ovarian epithelial tumors (p < 0.001 for both) and both markers were strongly correlated to each other (r = 0.877 & p < 0.001). Malignant tumors show significantly higher mean CD3+ T-lymphocytes than benign and borderline tumors. The mean CD3+ T-lymphocytes decrease significantly on increasing malignant tumor grade (p = 0.019) and expression of both FasL and survivin (r = −0.729, -0.582, respectively & p < 0.001 for both). The higher expression of FasL and survivin in malignant as compared to benign ovarian tumors suggest that they have a significant role in pathogenesis of ovarian carcinoma. Both markers are strongly correlated to each other and may contribute to immune escape of ovarian carcinoma as their higher expression is associated with decreased number of CD3 + T-lymphocytes.