妊娠滋养细胞疾病中NF-κBp65和IκBα的表达及临床意义

目的:分析核转录因子κB p65(NF-κBp65)和核转录因子κB抑制蛋白α(IκBα)在正常早孕绒毛及妊娠滋养细胞疾病中表达的差异性,以及与妊娠滋养细胞肿瘤(GTN)(包括侵蚀性葡萄胎和绒毛膜癌)患者年龄及临床分期的关系。并探讨两者在妊娠滋养细胞肿瘤组织中的相关性。方法:采用免疫组化方法(SP法)检测20例正常早孕绒毛组织、30例葡萄胎组织、13例侵蚀性葡萄胎和15例绒毛膜癌中NF-κBp65和IκBα的表达情况。结果:NF-κBp65在正常早孕绒毛组与侵蚀性葡萄胎组、正常早孕绒毛组与绒毛膜癌组、葡萄胎组与侵蚀性葡萄胎组、葡萄胎组与绒毛膜癌组之间阳性表达率比较差异均具有统计学意义(P=0.013,0.018,P=0.026,0.035,P<0.05);IκBα在正常早孕绒毛组与侵蚀性葡萄胎组、正常早孕绒毛组与绒毛膜癌组、葡萄胎组与侵蚀性葡萄胎组、葡萄胎组与绒毛膜癌组之间阳性表达率比较差异均具有统计学意义(P<0.01)。NF-κBp65和IκBα在GTN中的表达与临床分期有关(P=0.043,0.042,P<0.05),与患者年龄无关。NF-κBp65与IκBα在GTN中呈负相关(r=-0.403,P=0.034,P<0.05)。结论:NF-κBp65上调、IκBα的下降或缺失可能与滋养细胞肿瘤的发生发展以及侵袭、转移有关,NF-κBp65、IκBα两者在妊娠滋养细胞肿瘤组织中的表达呈负相关。     

Beclin-1在妊娠滋养细胞疾病中的表达意义

目的探讨Beclin-1在妊娠滋养细胞疾病中的表达意义。方法收集正常绒毛、葡萄胎、侵蚀性葡萄胎、绒毛膜癌、上皮样滋养细胞肿瘤和胎盘部位滋养细胞肿瘤组织共150例,采用免疫组织化学方法检测并分析各组组织中Beclin-1蛋白的表达水平。结果 Beclin-1在绒毛膜癌和侵袭性葡萄胎中的表达水平显著高于葡萄胎和正常绒毛(P0.05);化疗耐药的妊娠滋养细胞肿瘤病例中Beclin-1高表达,且与妊娠滋养细胞肿瘤的分期无明显相关性(P0.05)。结论 Beclin-1可能在妊娠滋养细胞疾病的进展和化疗耐药中起重要作用,有望成为判断葡萄胎恶变的辅助指标和化疗耐药评估候选指标之一。     

妊娠滋养叶细胞疾病的超声诊断

妊娠滋养叶细胞疾病( gestational trophoblastic disease,GTD)是指一组来源于胎盘绒毛滋养细胞的疾病。包括葡萄胎、侵蚀性葡萄胎和绒毛膜癌等。可以认为这几种疾病之间有一定联系,良性葡萄胎可能延续发展,经侵蚀性葡萄胎至绒癌。萄葡胎亚洲国家发病率比欧洲或北美高3~10倍[1]。约10%~20%的葡萄胎患者有可能发展为侵蚀性葡萄胎。绒癌除上述途径恶化而来外,也可直接发生于葡萄胎、流产或足月妊娠分娩以后。其临床诊断有一定困难,主要依靠病史、临床表现、HCG测定、诊断性刮宫等项检查。早期诊断与及时化疗是治疗成功的关键。近年来,随着超声显像技术的应用和发展,妊娠滋养叶细胞疾病的诊断率明显提高。现将其超声诊断的研究综述如下。     

滋养细胞疾病雌、孕激素受体的研究

滋养细胞疾病有无ER、PR存在及其变化情况如何?尚未见文献报道。本文采用酶联亲和组化法对正常早孕绒毛、足月妊娠胎盘各10例,滋养细胞疾病28例的ER、PR进行测定,结果表明:①早孕绒毛、胎盘、葡萄胎、恶葡、绒癌ER、PR均可被测知,滋养细胞疾病较正常早孕绒毛、足月胎盘ER、PR为低。②滋养细胞     

早孕绒毛及妊娠滋养细胞疾病中癌基因表达的研究

目的:研究癌基因c-myc和N-ras在早孕绒毛及GTD中的表达。方法:地高辛标记的c-myc、N-ras裸核探针与石腊包埋的18例早孕绒毛及54例GTD组织进行原位杂交。结果:c-myc mRNA在早孕绒毛、葡萄胎、侵蚀性葡萄胎和绒癌中阳性表达率分别为27.8%、44.4%、55.5%和84.4%。N-ras mRNA在早孕绒毛、葡萄胎、侵蚀性葡萄胎和绒癌组织中阳性表达率分别为83.8%、88.9%、77.8%和44.4%。Ⅲ-Ⅳ期GTT中c-myc阳性表达率高于Ⅰ期。c-myc和N-ras在GTT组织中共同表达率为47.2%。结论:c-myc与GTD病变的发展及临床分期有关,N-ras为GTT发生的早期事件。     

p53在妊娠滋养细胞和肿瘤中的表达

目的：分析Ｐ５３过度表达与葡萄胎、恶性葡萄胎（恶葡）、绒毛膜癌（绒癌）的恶性程度及预后关系。方法：采用单克隆抗体免疫组化技术，对正常胎盘绒志及妊娠滋养细胞肿瘤组织，进行肿瘤抑制基因Ｐ５３表达检测。其中包括早期妊娠１０例，中期妊娠１０例，晚期妊娠１３例，葡萄胎４０例，恶葡９例，绒癌２０例；结果：正常胎盘Ｐ５３异常表达阴性，葡萄胎、恶葡、绒癌阳性表达率分别为４２．５％（１７／４０）、５５．５％（５／９     

妊娠滋养细胞病变中FHIT和PTEN蛋白的表达

目的　探讨抑癌基因FHIT、PTEN蛋白表达在妊娠滋养细胞疾病发生、发展中的作用及临床病理意义。方法　采用免疫组化S P法分别检测了 30例完全性葡萄胎、15例侵袭性葡萄胎、15例绒毛膜细胞癌中FHIT、PTEN的蛋白表达。结果　完全性葡萄胎中FHIT蛋白表达低于正常早孕绒毛 (P <0 0 1) ,FHIT蛋白表达与葡萄胎滋养细胞增生呈负相关关系。侵袭性葡萄胎和绒毛膜细胞癌中FHIT蛋白表达均低于完全性葡萄胎 (P <0 0 5 ,P <0 0 1)。侵袭性葡萄胎和绒毛膜细胞癌中核PTEN蛋白表达均低于完全性葡萄胎 (P <0 0 1,P <0 0 1)。绒毛膜细胞癌中核PTEN蛋白表达与临床分期呈负相关关系。结论　FHIT和PTEN蛋白表达下降或缺失可能参与了妊娠滋养细胞疾病的发生、发展过程。     

胎盘部位滋养细胞肿瘤中NF-κBp65、CD44v6、Kiss-1的表达及相关性

目的 探讨细胞核因子-κBp65(NF-κBp65)、CD44v6、Kiss-1在胎盘部位滋养细胞肿瘤中的表达及相关性.方法 采用免疫组化PV 6000两步法检测20例正常早孕绒毛、15例葡萄胎、15例绒癌、10例胎盘部位滋养细胞肿瘤(placent site trophoblastic tumor,PSTT)中NF-κBp65、CD44v6、Kiss-1的表达情况及相关性.结果 NF-κBp65、CD44v6分别在正常早孕绒毛、葡萄胎、PSTT、绒癌中表达呈升高趋势,差异有统计学意义(P1＜0.01、P2＜0.01).NF-κBp65、CD44v6在PSTT中表达高于正常早孕绒毛和葡萄胎中的表达,差异有统计学意义(P1＜0.01、P2＜0.01、P3＜0.01、P4＜0.05);NF-κBp65在PSTT和绒癌中表达差异有统计学意义(P＜0.01);CD44v6在PSTT和绒癌中表达差异无统计学意义(P>0.05).Kiss-1在正常早孕绒毛、葡萄胎、PSTT、绒癌中表达呈下降趋势,差异有统计学意义(P＜0.01).Kiss-1在PSTT中表达低于正常早孕绒毛和葡萄胎中的表达,差异有统计学意义(P1＜0.01,P2＜0.01);在PSTT和绒癌中表达差异无统计学意义(P>0.05).NF-κBp65和CD44v6呈正相关(r=0.356,P=0.005)和Kiss-1呈负相关(r=-0.527,P=0.000);CD44v6和Kiss-1呈负相关(r=-0.346,P=0.007).结论 NF-κBp65、CD44v6的高表达和Kiss-1的低表达可能在胎盘部位滋养细胞肿瘤的浸润转移中起重要作用.     

HMGB1和NF-κBp65在葡萄胎、绒癌中的表达变化及意义

目的研究早期妊娠、晚期妊娠的胎盘组织、葡萄胎和绒癌组织中高迁移率族蛋白1(HMGB1)和核因子-κBp65(NF-κBp65)的表达变化及意义,探讨HMGB1和NF-κBp65与滋养细胞疾病的关系。方法选择2010年6月至2013年12月潍坊市中医院和潍坊市人民医院收治的流产、剖宫产、葡萄胎和绒癌标本,应用免疫组织化学方法检测HMGB1及NF-κBp65在早期妊娠、晚期妊娠的胎盘组织、葡萄胎和绒癌组织中的表达变化。结果 1.HMGB1阳性颗粒主要定位于细胞质和细胞核中,主要表达于绒毛滋养细胞和血管内皮细胞和间质细胞。与早期和晚期妊娠绒毛组织相比较,葡萄胎和绒癌组织中HMGB1阳性着色强度均明显增强,差异有统计学意义(P0.001)。2.NF-κBp65阳性颗粒主要定位在细胞质中,主要分布于绒毛滋养细胞、绒毛间质、血管内皮细胞。与早期和晚期妊娠绒毛组织相比较,葡萄胎和绒癌组织中NF-κBp65阳性着色强度均明显增强,差异有统计学意义(P0.001)。3.HMGB1与NF-κBp65平均光密度值在葡萄胎中表达强度呈正相关(r=0.7499,P=0.000);在绒癌组织中表达强度呈正相关(r=0.7338,P=0.000)。结论 HMGB1及NF-κBp65高表达与葡萄胎、绒癌的发生相关;HMGB1和NF-κBp65在葡萄胎和绒癌组织中的表达趋势一致,两者线性相关分析为正相关,提示HMGB1高表达可增强NF-κBp65的表达,与肿瘤细胞的生长有关。     

妊娠滋养细胞疾病中拓扑异构酶Ⅱα与Ki-67的免疫组织化学研究

一、材料与方法1.病例选择 :复习解放军总医院、香港大学玛丽医院和河北医科大学附属医院病理科存档的妊娠滋养细胞肿瘤 ,选择资料完整的 76例。其中 ,伴有绒毛水肿的流产 15份、部分性葡萄胎 2 0份、完全性葡萄胎 2 0份、侵蚀性葡萄胎 13份和绒癌 8份。除侵蚀性葡萄胎和绒癌为子宫切除标本外 ,余均来源于刮宫标本。所有标本均经 4%甲醛固定 ,石蜡包埋。同时选择妊娠早期和妊晚期正常胎盘作为对照组。2 .免疫组织化学方法 :采用枸橼酸 微波 ＡＢＣ法 ,检测鼠抗人拓扑异构酶 (ｔｏｐｏｉｓｏｍｅｒａｓｅ ,Ｔｏｐｏ)Ⅱα(单抗Ｋｉ Ｓ1,1∶2 0…     

Decreased expression of Ras GTPase activating protein in human trophoblastic tumors.

The normally developing placenta undergoes extensive but regulated noninvasive cellular proliferation. Various proto-oncogenes and growth factors have been associated with the regulation of trophoblastic placental growth. Activation of some oncogenes and altered expression of growth factors have been demonstrated in trophoblastic tumors (hydatidiform mole and choriocarcinoma). The ras proto-oncogene plays a key role in the signal transduction cascade of activated growth factors, and is known to be activated or overexpressed in multiple tumor types. Ras GTPase activating protein (RasGAP), a major down-regulator of ras activity, is present at high levels in placenta. To assess the role that Ras-GAP plays in the development of trophoblastic tumors, we performed immunohistochemical analyses with anti RasGAP antibodies of normal placentas, hydatidiform moles, invasive moles, and malignant choriocarcinomas. Normal placentas and noninvasive hydatidiform mole displayed intense positive staining confined to trophoblasts, whereas no staining was observed in the trophoblasts of invasive moles or choriocarcinomas. Thus, there was an inverse correlation between expression levels of RasGAP protein and the invasive potential and malignant phenotype in human trophoblastic tumors. The data indicate that RasGAP may play a regulatory role in trophoblast proliferation and that abolishing its activity may be associated with malignant transformation of these cells.     

子痫前期患者胎盘滋养细胞血管生长抑素、血管内皮因子表达的临床意义

目的检测胎盘部位血管生长抑素（AS）、血管内皮生长因子（VEGF）表达的水平,探讨在子痫前期患者发病中的临床意义。方法取84例孕妇胎盘绒毛组织,其中正常妊娠组30例,轻度子痫前期23例,重度子痫前期组31例。采用免疫组织化学方法检测在AS、VEGF胎盘部位的表达,光镜下观察表达的分布特点、细胞染色强度和阳性细胞数。结果在子痫前期组绒毛滋养细胞AS表达强度与正常对照组比较显著增强（P〈0.05）,表达强度与子痫前期病情呈正相关（r=0.371 P〈0．05）。子痫前期组与正常对照组胎盘VEGF的表达分布特点相同,子痫前期组较正常对照组显著降低,差异有统计学意义（P〈0．05）,表达强度与子痫前期病情呈负相关（rs=-0．473P〈0．05）。对正常对照组及子痫前期组的AS、VEGF的表达进行组内相关性分析,显示As与VEGF之间存在关联关系,呈负相关。相关系数分别为r=-0．476P〈0．05,r=-0．419P〈0．05。结论AS和VEGF在子痫前期患者胎盘部位的表达有显著变化,两者之间呈显著负相关,可能是子痫前期发病的重要因素之一。     

P57kip2 immunohistochemical expression and ultrastructural findings of gestational trophoblastic disease and related disorders

Gestational trophoblastic disease (GTD) is a unique spectrum of diseases ranging from complete hydatidiform mole (CHM), partial hydatidiform mole (PHM), and invasive mole (IM) to choriocarcinoma (CC). Placental site trophoblastic tumor (PSTT) and epithelioid trophoblastic tumor (ETT) have been classified as related disorders. Mesenchymal dysplasia (MD) may be misdiagnosed as PHM; however, it is said to have a quite different histogenesis from PHM. P57kip2 is the protein product of a paternally imprinted or maternal gene that inhibits cyclin-dependent kinases (CDK), thus serving to inhibit cell proliferation and to suppress tumor growth. Its lack of expression in trophoblastic disease plays a role in its abnormal proliferation and differentiation. In this study, P57kip2 immunostaining was absent in the trophoblastic layers of CHM and was positive in the trophoblast layer of nonmolar villi and MD. Ultrastructure of complete molar cystic villi showed tree-like branching of microvillous processes and intracytoplasmic lacunae without capillaries in the stroma, whereas MD contained many newly formed blood vessels and collagen. Also, large lacunae with microvilli and polymorphic nuclei of syncytiotrophoblast cells with well-developed organelles were observed in IM. Lung ETT following CHM and normal deliveries showed two types of large mononuclear cells and binuclear cells with abundant organelles and bundles of intermediate-type filaments in the stroma.     

激光捕获显微切割技术联合短串联重复序列多态性分析在葡萄胎受精类型鉴别中的应用研究

目的探讨激光捕获显微切割技术（LCM）联合短串联重复序列（STR）多态性分析用于葡萄胎受精类型鉴别的可行性。方法首先通过LCM获得葡萄胎病病理组织的滋养细胞,再用16个位点（D8S1179,D21S11,D2S1338,CSF1PO,TH01,TPOX,D3S1358,D13S317,D16S319,D19S433,FGA,D18S51,D5S818,VWA,D7S820,Am）复合微卫星序列-PCR方法对61例病理学诊断为葡萄胎标本进行分析,明确其受精类型。结果用LCM技术从61例标本中获得滋养细胞后,成功提取DNA46例,浓度范围为：6.2-114.5（ngμ/l）。经STR多态性分析确定空卵单精子受精完全性葡萄胎13例,空卵双精子受精完全性葡萄胎13例,部分性葡萄胎20例。结论 LCM联合多重STR多态性分析能够确定葡萄胎的受精类型。     

Complete hydatidiform mole in twin pregnancy

Six cases of hydatidiform mole associated with normal chorionic villi and a normal embryo/fetus (in five cases) were investigated with interphase cytogenetic and DNA cytometric analyses for diagnostic purposes. DNA probes specific for the pericentromeric regions of chromosomes 1 and X and for the long arm of chromosome Y were used. In four cases a dizygotic twin pregnancy could be proven. In these cases, the histologically normal chorionic villi showed an XY DNA-diploid pattern, consistent with a normal male conceptus, and the molar chorionic villi a XX pattern. In the other two cases an identical sex chromosomal pattern was found in the normal and in the molar villi (XX/XX and XY/XY respectively). In all six cases the molar placental tissues showed prominent trophoblastic hyperplasia with DNA-polyploidy, consistent with a complete hydatidiform mole. In two cases persistent gestational trophoblastic disease developed. It is emphasized that twin pregnancies composed of a normal conceptus and a complete mole have a relatively high risk for the development of persistent trophoblastic disease and therefore, should be carefully differentiated from triploid partial moles with a relatively low risk of persistent gestational trophoblastic disease. These case reports indicate that additional interphase cytogenetic and DNA cytometric analyses are useful in this differential diagnosis.     

子宫内膜样腺癌中Maspin、VEGF—C的表达与肌层浸润程度的临床研究

目的检测maspin和VEGF—C在子宫内膜样腺癌中的表达及与肌层浸润程度的意义。方法采用免疫组化法检测49例子宫内膜样腺癌及15例正常子宫内膜中maspin、VEGF—C的表达及与肌层浸润程度的意义。结果（1）Maspin在子宫内膜样腺癌中表达阳性,肌层浸润深maspin阳性表达率增高,差异无统计学意义（P〉0．05）。（2）VEGF—C在子宫内膜样腺癌中表达阳性,肌层浸润深度〉1／2组阳性率（75．0％）高于肌层浸润深度≤1／2组（36．4％）,差异具有统计学意义（P〈0．05）。（3）子宫内膜样腺癌中maspin和VEGF—C呈正相关关系（P〈0．05）。结论maspin和VEGF—C表达上调可能共同促进了子宫内膜样腺癌的发生、发展、侵袭及转移,二者联合检测可能对临床诊疗、判断预后有指导意义。     

妊娠滋养细胞肿瘤中E-cadherin和增殖细胞核抗原的表达

目的：研究妊娠滋养细胞肿瘤(GTT)中E-cadherin与PCNA的表达意义。方法：采用免疫组化SP法检测了妊娠滋养细胞肿瘤中E-cadherin与PCNA的表达。结果：E-cadherin在正常早期绒毛(NP)及其肿瘤(CM、IM、OCA)中表达率分别为56．29％、42．07％、19．30％、7．14％,各组间差异非常显著。PCNA在NP、CM、IM和OCA中表达率分别为10．40％、20．76％、53．60％、51．95％,各组间差异非常显著。E-cadherin与PCNA阳性率呈负相关。结论：E-cadherin和PCNA的检测有助于良恶性GTT的鉴别;E-cadherin表达减弱或消失促进了GTT的增殖活性。     

p57(KIP2) immunohistochemical staining of gestational trophoblastic tumours does not identify the type of the causative pregnancy

AIM: To determine whether immunohistochemical staining for p57(KIP2), the product of the maternally expressed gene CDKN1C, can be used to differentiate between gestational trophoblastic tumours arising from a complete hydatidiform mole and those originating from non-molar pregnancies. METHODS: The immunohistochemical expression of p57(KIP2) was investigated in 23 cases of choriocarcinoma and 17 placental site trophoblastic tumours. Fourteen of the tumours examined were shown by DNA analysis to have arisen from complete hydatidiform moles and 26 from non-molar pregnancies. RESULTS: Five of 11 (45%) post-complete hydatidiform mole choriocarcinomas and two of three (67%) post-complete hydatidiform mole placental site trophoblastic tumours were found to be p57(KIP2)+ and showed similar immunostaining characteristics to tumours that developed from non-molar pregnancies. Although there was a statistically significant reduction in the proportion of cases showing positive p57(KIP2) staining in post-complete hydatidiform mole tumours compared with those originating in non-molar pregnancies [proportion difference 0.35 [95% confidence interval (CI) 0.05, 0.61], P = 0.02], immunostaining did not provide diagnostically useful information to differentiate between these tumours in clinical practice. There was no significant difference between the extent of staining in choriocarcinoma versus placental site trophoblastic tumours [proportion difference 0.17 (95% CI - 12, 42), P = 0.19]. The majority of both types of gestational trophoblastic tumour were positive for the presence of the p57(KIP2) protein irrespective of their genetic origin. CONCLUSION: Immunostaining for p57(KIP2) fails to discriminate between gestational trophoblastic tumours that have arisen from complete hydatidiform moles and those that have originated from other types of pregnancy.     

妊娠期高血压疾病患者胎盘组织中抵抗素的表达

目的检测抵抗素（Resistin）在妊娠期高血压疾病患者胎盘组织中的表达,探讨抵抗素在妊娠期高血压疾病发病机制中的作用。方法选取妊娠期高血压疾病患者40例,包括妊娠期高血压患者20例,子痫前期患者20例,选取无高血压、糖尿病等并发症的孕妇20例作为对照组。并记录每个患者新生儿出生体重。应用免疫组化SP两步法检测抵抗素在其胎盘组织中的表达情况,对免疫组化切片采用Tanaka等级评分法进行评分。应用studengt t检验,分别对子痫前期组与对照组,妊娠期高血压组与对照组免疫组化评分结果进行比较;并采用双变量相关法,估计胎盘抵抗素的表达与新生儿出生体重的相关性,以P〈0.05为差异有显著性。结果本研究发现抵抗素表达于胎盘绒毛滋养细胞的细胞浆中;对免疫组化切片应用Tanaka等级评分法进行评分,经studengt t检验发现子痫前期组抵抗素在胎盘组织中的表达低于对照组,P〈0.05,差异有显著性;妊娠期高血压组与对照组差异无显著性,P〉0.05。抵抗素在胎盘组织中的表达与新生儿出生体重正相关（r=0.883,P〈0.05）。结论抵抗素可能参与了子痫前期的发病机制,推测其表达下降是由胎盘功能减退引起的,这可能是疾病发展过程中的一个结果而不是原因;抵抗素可能参与了子痫前期胎儿生长受限的发生,其具体机制将有待于进一步研究。     

子痫前期患者胎盘组织中尾加压素Ⅱ及一氧化氮的表达及意义

目的研究尾加压素Ⅱ（urotensin Ⅱ,UⅡ）及一氧化氮在子痫前期患者胎盘组织的表达情况及其与子痫前期发病的关系。方法采用RT-PCR方法检测45例子痫前期患者（轻度22例,重度23例）及20例正常晚期妊娠孕妇胎盘组织中UIImRNA的表达情况;同时用硝酸还原酶法检测子痫前期组孕妇与正常组孕妇胎盘组织中NO浓度。结果重度子痫前期患者胎盘组织UIImRNA（0.85±0.40）明显高于正常妊娠组（0.38±0.30）（P〈0.05）;轻度子痫前期患者胎盘组织UIImRNA（0.64±0.31）,与正常妊娠组比较差异无统计学意义（P〉0.05）。轻、重度子痫前期患者胎盘组织NO浓度分别为114.42±6.52 u/mg、79.22±3.31 u/mg,均显著低于正常妊娠组241.36±13.24u/mg（P〈0.05）。结论子痫前期胎盘组织中尾加压素Ⅱ水平升高,NO浓度下降,可能在子痫前期的发病中起一定作用。