抑癌基因P16在卵巢肿瘤组织中的表达

抑癌基因ｐ１６在卵巢肿瘤组织中的表达杨红郑维国辛晓燕陈必良本研究应用免疫组化ＡＢＣ法及原位杂交方法，对７２例卵巢肿瘤及正常卵巢组织中的ｐ１６蛋白和ｍＲＮＡ进行了检测，并初步探讨了抑癌基因ｐ１６与卵巢癌发生、发展及生物学行为的关系。现报道如下。一、资料...     

Survivin Ki67在卵巢原发癌与转移癌中的表达及其临床意义

目的研究卵巢原发癌和转移癌组织中SurvivinmRNA、Ki67蛋白表迭的相关关系及临床意义。方法对1985—2002年中国医科大学附属一院妇科收治的38例卵巢原发癌、36例卵巢转移癌、16例卵巢良性肿瘤病例组织标拳及10例正常卵巢组织运用原住杂交技术检测SurvivinmRNA的表达．免疫组化SP法检测Ki67蛋白标记指数，并对其与临床各因素关系进行综合分析。结果Survivin、Ki67在卵巢原发癌和转移癌组织中的表达水平明显高于正常卵巢及良性卵巢肿瘤组织（P〈0．01）。Ki67表达与年龄及有无淋巴结转移无关，与卵巢原发癌和转移癌的临床分期、分化程度及卵巢转移癌的预后密切相关。Survivin表达与K167表达具有一定关联性，呈正相关关系（r=0．498）。结论Survivin和Ki67的高表达在卵巢原发癌厦转移癌的发生发展中有协同作用，可作为评价卵巢肿瘤生物学行为的重要依据，并可评估卵巢肿瘤恶性程度的重要参考指标。     

肿瘤坏死因子基因在卵巢肿瘤中的表达

以地高辛配基记的肿瘤坏死因子－ａｃＤＮＡ为探针，采用原位杂交技术对５４例卵巢肿瘤及１０例正常卵巢组织新鲜标本制成的冰冻切片，进行肿瘤坏死因子基因检测，观察卵巢肿瘤细胞及肿瘤浸润淋巴细胞ＴＮＦ基因的表达。结果：卵巢癌细胞及ＴＩＬ的ＴＮＦ基因阳性率明显高于卵巢交界性和良性肿瘤的瘤细胞及其ＴＩＬ；临床分期赵晚，癌细胞内ＴＮＦ基因的阳性纺赵高，而ＴＩＬ内ＴＮＦ基因阳性率赵低，表明：卵巢肿瘤细胞及ＴＩＬ内Ｔ     

基于组织连续切片子宫骶骨韧带内神经脉管数字化三维模型构建研究

目的 研究基于新鲜组织连续切片数据集构建子宫骶骨韧带内神经脉管的数字化三维模型。方法 选择2014年8月于南方医科大学南方医院因宫颈鳞状细胞癌ⅠB1期行经腹广泛性子宫切除术的新鲜右侧子宫骶骨韧带标本1例,标本外侧缘取切片经病理学检查证实无癌细胞浸润。用印度墨水标记韧带宫颈侧与上缘,标本常规固定、脱水后,利用穿刺针穿取4条略长于标本的猪肝组织（直径约0.5mm）作为定位杆标记,与标本一起进行石蜡包埋,从宫颈侧开始连续切片,切片厚度5μm,连续5张为一组分别进行HE染色、动脉弹力纤维特殊染色及免疫组织化学特殊染色。对所有连续切片进行图像采集、融合,并对其进行配准。利用计算机三维重建软件构建出子宫骶骨韧带内神经与脉管数字化三维模型。结果 成功利用组织连续切片数据集构建出子宫骶骨韧带及其内神经与脉管的数字化三维模型。结论 通过观察重建的数字化三维模型,可清楚地显示出交感神经、副交感神经、血管及淋巴管在子宫骶骨韧带内的走行分布及立体关系,为系统保留盆腔自主神经的广泛性子宫切除术步骤的规范化提供依据。     

钙黏素6基因在卵巢肿瘤组织中的表达及突变的研究

目的　通过研究卵巢肿瘤组织中钙黏素 (cadherin) 6基因的表达、突变及其临床意义 ,以寻找与卵巢肿瘤发生相关的分子水平标志物。方法　应用RT PCR技术和PCR 单链构象多态性方法分别检测恶性卵巢肿瘤组织 (4 1份 )、良性卵巢肿瘤组织 (15份 )、正常卵巢组织 (17份 )中cadherin 6mRNA的表达及cadherin 6基因的突变情况 ,并分析其临床意义。结果　正常卵巢组织、良性卵巢肿瘤组织、恶性卵巢肿瘤组织中cadherin 6mRNA阳性率分别为 71%、5 3%、2 4 % ,前两者显著高于后者 (P<0 0 5 )。cadherin 6mRNA阳性率与恶性卵巢肿瘤手术病理分期有关 ,Ⅲ～Ⅳ期恶性卵巢肿瘤组织中 ,cadherin 6mRNA阳性率 (5 % )显著低于Ⅰ～Ⅱ期 (4 5 % ,P <0 0 1)。 4 1例恶性卵巢肿瘤患者中 ,2例出现cadherin 6基因突变 ,而在正常卵巢组织和良性卵巢肿瘤组织中无cadherin 6基因突变。结论　cadherin 6mRNA在晚期恶性卵巢肿瘤组织中表达缺失。提示cadherin 6mRNA表达可作为判断恶性卵巢肿瘤预后的一个指标     

妊娠合并卵巢肿瘤诊疗进展

妊娠合并卵巢肿瘤是少见疾病,由于涉及母婴两方面,诊断和治疗与普通卵巢肿瘤不同。对肿瘤性质进行正确鉴别诊断是治疗的基础,也是降低不良妊娠事件发生率的前提。超声和磁共振成像(MRI)在妊娠期卵巢肿瘤的鉴别诊断中有重要作用。妊娠期化疗、妊娠期腹腔镜手术也为妊娠期卵巢恶性肿瘤的治疗提供新思路。总结近年妊娠合并卵巢肿瘤诊疗进展,为鉴别诊断及治疗提供参考。     

应用组织芯片技术研究卵巢癌组织中缺氧诱导因子1α与血管生成的关系

目的　探讨缺氧诱导因子 1α(hypoxiainduciblefactor 1α ,HIF 1α)在卵巢癌组织中的表达及其与血管生成的关系。方法　联合应用组织芯片和原位杂交、免疫组化技术 ,检测 2 95份卵巢上皮性肿瘤 (其中卵巢癌 2 38份、交界性卵巢肿瘤 19份、良性卵巢肿瘤 38份 )及 13份正常卵巢组织中HIF 1αmRNA和血管内皮生长因子 (VEGF)的表达情况 ,以CD3 4 单克隆抗体标记血管内皮细胞来检测微血管密度 (MVD)。结果　卵巢癌、交界性卵巢肿瘤和良性卵巢肿瘤、正常卵巢组织中HIF 1αmRNA的阳性表达率分别为 81 9%、4 2 1%、13 2 %和 0。交界性卵巢肿瘤和卵巢癌组织中HIF 1αmRNA的表达高于正常卵巢和良性卵巢肿瘤 ,卵巢癌高于交界性肿瘤 ,差异均有统计学意义 (P <0 0 1)。卵巢癌组织中HIF 1αmRNA的表达与手术病理分期和病理类型无关 (P >0 0 5 ) ,而与病理分级呈正相关 (r=0 2 4 6 ,P <0 0 1)。卵巢癌组织中HIF 1αmRNA表达与VEGF表达 (r =0 2 0 6 ,P =0 0 1)和MVD计数 (r =0 4 5 1,P <0 0 1)均呈显著正相关。结论　卵巢癌组织过度表达HIF 1αmRNA ,并通过诱导VEGF促进肿瘤的新生血管形成。     

耐药相关基因在卵巢癌组织中的表达及其临床意义

目的 通过测定多药耐药基因（ＭＤＲ１,多药耐药相关蛋白（ＭＲＰ）,肺耐药相关蛋白（ＬＲＰ）和谷胱甘肽Ｓ－转移酶（ＧＳＴ－π）的卵巢癌组织中的表达,了解其在卵巢癌化学治疗耐药中的作用。方法 采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）测定了４１例卵巢癌,２５例良性卵巢肿瘤和１２例正常卵巢组织中ＭＤＲ１,ＭＲＰ,ＬＲＰ和ＧＳＴ－π的表达,并分析其与临床,病理特征的关系。结果 （１）在正常卵巢,良性卵巢肿瘤     

MEX3A蛋白在上皮性卵巢癌组织中的表达及临床意义

目的:探究MEX3A蛋白在上皮性卵巢癌组织中的表达水平及其与患者临床病理特征之间的关系。方法:免疫组化法检测46例上皮性卵巢癌、25例良性卵巢肿瘤及15例正常卵巢组织中MEX3A蛋白表达。结果:MEX3A蛋白在上皮性卵巢癌组织中的阳性表达明显高于良性卵巢肿瘤和正常卵巢组织(P0.05),而良性卵巢肿瘤和正常卵巢组织比较则无显著差异(P0.05)。MEX3A蛋白表达与上皮性卵巢癌的临床分期、淋巴结转移有关(P0.05),而与患者年龄、组织分级、组织类型及有无腹水无关(P0.05)。结论:MEX3A蛋白在上皮性卵巢癌组织中高表达,并与患者临床分期和淋巴结转移相关,在卵巢癌的发生发展中具有重要作用。     

卵巢肿瘤组织中c-met蛋白表达的意义

目的：探讨肝细胞生长因子（HGF）受体c-met蛋白在卵巢肿瘤组织中的表达及其与细胞增生和血管形成之间的关系。方法：应用免疫组化SABC法检测34例卵巢恶性肿瘤、15例卵巢交界性肿瘤和20例卵巢良性肿瘤组织的c-met蛋白的表达及其增殖细胞核抗原（PCNA）指数和微血管密度（MVD）。结果：c-met在卵巢良性，交界性和恶性肿瘤组织之间的表达有显著性差异，且有逐渐增高趋势。在卵巢恶性肿瘤组织中c-met蛋白表达与PCNA指数及MVD数值相关，c-met蛋白表达与卵巢恶性肿瘤病理分化之间密切相关。结论：HGF和c-met可能促进细胞增殖和血管形成的途径而在卵巢恶性肿瘤的发生，发展过程中起重要作用。     

Uptake of a cholesterol-rich emulsion by neoplastic ovarian tissues.

OBJECTIVE: Previously, it was shown that a lipidic emulsion (LDE) composed of phospholipids and cholesterol esters which binds to low-density lipoprotein (LDL) receptors may concentrate in acute myeloid leukemia cells. In this study, we aimed to verify whether LDE also has the ability to concentrate in malignant ovarian cancer after being injected into the blood circulation of the patients. METHODS: Three groups of women scheduled for surgery were included in the survey: 13 bearing malignant tumors, 9 with benign ovarian tumors, and 13 without ovarian tumor who were scheduled to undergo oophorectomy due to malignant disease of the uterine cervix or endometrium. On the day prior to surgery they were injected with LDE labeled with [(14)C]cholesteryl oleate. Specimens of tumors and normal ovaries excised during surgery were lipid extracted and analyzed for radioactivity counting. Results were expressed in radioactive count (cpm) per gram of tissue. RESULTS: The mean of the uptakes of the emulsion radioactivity by the malignant tumors was roughly eightfold greater when compared with that of the contralateral normal ovaries (2261 +/- 1444 and 275 +/- 137 cpm/g, respectively, P < 0.012), benign tumors, and normal ovaries of the patients without ovarian tumors. CONCLUSION: LDE has the ability to concentrate in malignant ovarian tumor tissue. Therefore, it can be used as a vehicle to direct cytotoxic drugs against malignant ovarian tumors, thus diminishing the side effects of chemotherapy.     

Expression of epidermal growth factor receptors in benign ovarian tumors

The objective of the present study was to determine the level of EGFR expression in patients having benign ovarian tumors and in morphologically normal ovarian tissue by quantitative ligand binding assays. In a prospective study 42 patients with benign ovarian tumors and 30 with normal ovaries were included. In the group of patients expressing EGFR, those having benign tumors were 23 (64%) and the ones with normal ovaries--13 (36%). The average level of EGFR expression was the highest in the cases of teratoma adultum (110 fmol/mg), followed by mucinous (97 fmol/mg) and serous (30 fmol/mg) cystadenomas. The differences between the average levels of EGFR expression in mucinous and serous benign ovarian tumors were significant (p = 0.032), as for the remaining histological variants--statistically insignificant (p = 0.086). Mucinous cystadenomas possessed higher proliferative potential compared to serous cystadenomas.     

Three-dimensional ultrasound and three-dimensional power Doppler in the assessment of adnexal masses

Three-dimensional ultrasound is a new, emerging technology that provides additional information for the evaluation of ovarian tumors. Multiplanar and volume-rendering display methods combined with the ability to rotate volume data into standard orientations are essential components of the current and future success of three-dimensional ultrasound.

Increasing knowledge about three-dimensional ultrasound, as well as improved handling allow application of this method to the field of gynecological oncology. The recent development of real-time three-dimensional ultrasound imaging will further advance the clinical applications, particularly in the assessment of pelvic tumors. The introduction of the three-dimensional power Doppler systems may improve the information available on ovarian tumor vascularity and speed up the entire patient management process.     

Insulin producing primary ovarian carcinoid tumor

Carcinoid tumors are slow growing and originate most frequently from gastrointestinal tissue 1. They may also appear in genital tissue like the ovaries. Primary ovarian carcinoid tumors are rare and contribute to less than 0.1% of all ovarian carcinomas 2. We report the first case of insulin producing primary carcinoid tumor of the ovary, initially presented with amnesia and hypoglycemia and subsequently successfully treated with surgery.     

Preliminary observations on whole-ovary xenotransplantation as an experimental model for fertility preservation

Ovarian tissue preservation and retransplantation is a promising strategy to restore fertility in cancer survivors. Ischaemia accompanying ovarian tissue grafting, however, can lead to significant follicle loss. Transplantation of the whole ovary by vascular anastomosis has been considered as an alternative to prevent widespread ischaemic damage. In this study, the feasibility and function of transplanting whole ovary with intact vasculature were evaluated, with the goal of developing a xenograft model for studies using donated human ovaries. Whole-swine ovaries with vascular pedicles were perfused and transplanted as intact ovaries by anastomosis into irradiated ovariectomized nude rats (n = 10). The observation period was between 1 and 4 weeks. Fresh swine ovaries served as controls (n = 10). Ovarian stroma and follicle populations were assessed through histological examination in both transplanted and control ovaries. Most of the transplanted whole ovaries (n = 6) maintained stromal quality and all preantral follicle classes were represented, although follicle numbers decreased compared with fresh control. Four transplanted ovaries were fibrotic after 1–4 weeks within the nude rat. Our results demonstrate transplantation of whole-pig ovary into nude rats is possible and support development of this xenograft model system for human studies.     

Absence of estrogen receptor-beta expression in metastatic ovarian cancer

OBJECTIVE: To evaluate the expression of estrogen receptor (ER)alpha and ERbeta mRNA and protein in normal ovarian tissue and primary and metastatic tumors. METHODS: Estrogen receptor alpha and ERbeta expression was studied in normal ovarian biopsies (n = 9) and primary (n = 8) and metastatic ovarian epithelial cancers (n = 8). Ovarian tissue was collected from surgical samples. Estrogen receptor alpha and ERbeta mRNA expression was compared by coamplification of the mRNA of the ERs. Expression was confirmed at the protein level by Western blot analysis using antibodies specific for each receptor. RESULTS: Among eight primary ovarian cancer samples, three had only ERalpha, two had only ERbeta, and three had both. All eight metastatic ovarian cancer tissues expressed only ERalpha mRNA and protein. Biopsies from normal ovaries had ERalpha and ERbeta mRNA and protein. Two of the ovarian epithelial cancer samples were paired and showed the same results. CONCLUSION: We found varying amounts of ERalpha and ERbeta in normal ovaries, lower levels of ERbeta expression in ovarian epithelial cancer primary tumors, and only ERalpha in metastatic tumors. Our findings indicate that a fundamental difference might exist between primary and metastatic cells, which could be caused by intrinsic or extrinsic factors that regulate ER gene expression.     

Cyclin D1 overexpression and p53 mutation status in epithelial ovarian cancer

OBJECTIVE: To examine the cyclin D1 mRNA expression level in ovarian tumor samples as compared with normal ovaries and to determine the relationship between cyclin D1 overexpression and p53 mutation status in ovarian tumors. METHODS: mRNA was isolated and cDNA was prepared from 27 epithelial ovarian tumors (3 tumors of low malignant potential (LMP) and 24 cancers) and 6 normal ovaries. The cyclin D1 sequences were amplified by using a thermal cycler in parallel with the beta-tubulin gene as an internal control. The cyclin D1 mRNA expression level relative to beta-tubulin was determined by 32P phosphoimager analysis. To confirm the overexpression of the cyclin D1 protein in ovarian tumor cells, immunostaining was performed. The p53 gene mutation status was examined by direct cDNA sequencing. RESULTS: mRNA levels of cyclin D1 were significantly higher in 21 (78%) of the 27 ovarian tumors than in normal ovaries. Cyclin D1 overexpression was detected in ovarian LMP tumors as well as in ovarian cancer cases. Positive immunostaining of cyclin D1 protein was observed in 10 of 18 (56%) ovarian tumors examined. p53 mutations were found in 11 (61%) of 18 ovarian tumors. Of 11 ovarian tumor cases with p53 mutations, 5 showed overexpression of cyclin D1. All 7 ovarian tumor cases without p53 mutations showed significant cyclin D1 mRNA overexpression. CONCLUSION: Cyclin D1 overexpression seems to be an early genetic event in ovarian tumor development. Although p53 may be one of the proteins whose function regulates the expression of G1 cyclins, ovarian tumors with no p53 mutation consistently showed cyclin D1 overexpression. Cyclin D1 overexpression may play an important role in the tumorigenesis of epithelial ovarian tumors.     

Nuclear survivin expression is a positive prognostic factor in taxane-platinum-treated ovarian cancer patients

Objective

Study of the hen immune system led to seminal contributions to basic immunological principles. Recent studies of spontaneous ovarian cancer in the laying hen show strikingly similar tumor types and antigen expression compared to human ovarian cancer, suggesting hens would be valuable for studies of tumor immunology and pre-clinical vaccine development. Circulating mesothelin is a relatively specific marker for human ovarian cancer and autoantibodies to mesothelin were reported. We hypothesized that hen tumors express mesothelin and that circulating anti-mesothelin antibodies occur in response to tumors.

Methods

Mesothelin mRNA expression was analyzed by RT-PCR in hen ovarian tumors and normal ovaries. Mesothelin protein expression was evaluated by immunohistochemistry (IHC) and two-dimensional SDS-PAGE Western blots. Anti-mesothelin antibodies were assessed by immunoassay of sera from hens with normal ovaries and with ovarian tumors.

Results

Significant mesothelin mRNA expression was observed in 57% (12/21) of hen ovarian tumors but not in normal ovaries and was found predominantly in serous tumors as in humans. Mesothelin protein was detected in tumors with mesothelin mRNA by IHC and 2D Western blots, but not in normal ovaries or tumors without mesothelin mRNA. Circulating anti-mesothelin antibodies occurred in 44% (n = 4/9) of hens with ovarian tumors which express mesothelin mRNA and were not found in hens with tumors that did not express mesothelin (n = 0/5) or normal ovaries (n = 0/5).

Conclusion

The results support the utility of the hen as a novel model for preclinical studies of mesothelin as a biomarker and a target for immunotherapy.     

B7-H4 is over-expressed in early-stage ovarian cancer and is independent of CA125 expression

OBJECTIVE: This study characterizes the expression of the novel biomarker B7-H4 in ovarian cancer tissue, normal ovaries, and benign ovarian tumors, and evaluates its relationship to CA125. METHODS: Ovarian tissue lysates from 251 patients with ovarian carcinoma were assessed for the levels of B7-H4 and CA125 by ELISA assays. For comparison, ovarian tissues from patients with benign ovarian tumors (n=43) and patients with normal ovaries (n=32) were tested. The marker concentrations were correlated with CA125 expression, clinicopathological variables, and patient outcome. RESULTS: Using a cut-off based on the 95th percentile of B7-H4 or CA125 concentration in the control group, B7-H4 was over-expressed in 48% of patients with stage I cancer, 55% of patients with stage II cancer, and 67% of patients with late stage cancer. CA125 was elevated in 31% patients with early stage cancer. B7-H4 was elevated in tumors of 30 patients with early stage cancer that were negative for CA125. The combination of B7-H4 and CA125 identified 56 early stage cancer patients (65%) as positive. Correlation of marker expression to clinical outcome showed that high B7-H4 levels were correlated with poor prognosis. However, the effect was not significant when outcome was adjusted for other clinicopathological variables. CONCLUSION: B7-H4 expression was low in normal ovaries and in benign tumors while half of early stage and two-thirds of late stage cancers over-expressed B7-H4. The data are consistent with previous observations and support further investigation of B7-H4 in the detection of early stage ovarian cancer either alone, or in combination with CA125.     

Thymidine phosphorylase activity and neo-angiogenesis in ovarian cancer

Neo-angiogenesis seems to play an important role in the progression of ovarian cancer and in formation of distant metastases. Data from literature on role of phosphorylase in neoplasmatic disease and in neo-angiogenesis are controversial. In mammalian cytosole there are two different pirymidine nucleosyde phosphorylases: thymidine (PT) and uridine (PU). Both of them play important role in the metabolism of nucleosides as well as in the recycling of pyrimidine base. Recently thymidine phosphorylase is identified with platelet derived endothelial cell growth factor (PD-ECGF). It has been demonstrated, that PD-ECGF/PT influence on neo-angiogenesis and correlates with degree of neoplasmatic invasion. In literature the data about thymidine phosphorylase activity and its correlation with neoplasmatic angiogenesis in ovarian tumors are controversial. The aim of the study was to evaluate the activity of PT together with the intensity of angiogenesis in epithelial ovarian tumors. 42 patients with ovarian cancer were included into the study. The enzyme activity was measured in ovarian cancer tissue and in the serum in the spectrophotometer. Intratumoral microvessel density (IMD) was evaluated in tumor using immunohistochemical methods. 10 woman with normal ovaries, treated surgically due to non-oncological reasons served as a control. Activity of PT in ovarian tumor and in serum was compared to the control group. Correlation between the intensity of angiogenesis and PT activity in ovarian cancer was also investigated. Significantly higher PT activity was stated both in tumor and serum when compared to the control. Positive correlation between enzyme activity in the serum and neoplasmatic tissue was found. Surprisingly, the negative correlation between neo-angiogenesis and PT activity in ovarian cancer was observed. Neo-angiogenesis is higher in ovarian cancer, when compared to the group of borderline malignancy tumors. Positive correlation between PT activity and staging in ovarian cancer was observed. No correlation between grading and histopathological type of epithelial cancer was observed. PT activity and neo-angiogenesis evaluation might be useful in diagnostics of ovarian cancer.