MRI诊断妊娠滋养细胞疾病的临床价值

妊娠滋养细胞疾病(gestational trophoblastic dis-ease,GTD)是一组起源于胎盘滋养细胞的疾病,主要有葡萄胎和妊娠滋养细胞肿瘤(gestational trophoblastic neoplasia,GTN)[1,2]。2002年世界卫生组织(WHO)对女性生殖系统组织学分类进行修改,2003年正式公布GTD组织学新分类,指出GTD包括完全性葡萄胎、部分性葡萄胎、侵袭性葡萄胎、绒癌、     

《FIGO2015妇癌报告》解读连载七——妊娠滋养细胞疾病诊治指南解读

         妊娠滋养细胞疾病（GTD）是一组与异常妊娠相关的罕见疾病。包括良性的部分性葡萄胎和完全性葡萄胎,以及恶性侵蚀性葡萄胎和转移性葡萄胎、绒癌、胎盘部位滋养细胞肿瘤（PSTT）和上皮样滋养细胞肿瘤（ETT）。葡萄胎排空后可能发生人绒毛膜促性腺激素（HCG）持续升高（完全性葡萄胎15％～20％,部分性葡萄胎0.1％～5％）,也可能进展为绒癌。恶性GTD也称为妊娠滋养细胞肿瘤（GTN）。
         葡萄胎在亚洲一些地区较常见,发病率高达2/1000妊娠。欧洲和北美发病率通常小于1/1000妊娠。近年来,亚洲国家葡萄胎的发生率似乎在减少,可能与经济和饮食的改善以及出生率下降相关。绒癌的发病率很难估算,因其罕见,并且临床上由于缺乏组织病理学证据,很难将发生于葡萄胎的绒癌与侵蚀性葡萄胎区分开来。据报道绒癌发病率大约占妊娠的1/4 0000~9/4 0000,发病率一直在下降。PSTT和ETT比绒癌更罕见。     

妊娠滋养细胞疾病诊断与治疗指南(第四版)

正妊娠滋养细胞疾病(GTD)是一组来源于胎盘滋养细胞的疾病。从流行病学来看,葡萄胎在我国及亚洲一些地区较常见,发病率高达2/1000次妊娠;欧洲和北美发病率通常小于1/1000次妊娠。近年来,亚洲国家葡萄胎的发生率有所下降,可能是经济和饮食的改善以及生育率下降所致。绒毛膜癌(绒癌)的发病率低,难以估算,约为(1~9)/40 000次妊娠,由于临床上缺乏组织病理学证据,发生于葡萄胎后的绒癌难以与侵蚀性葡萄胎     

妊娠滋养细胞疾病和正常胎盘中EGFR相关产物表达及与葡萄胎后持续性滋养细胞肿瘤发生的关系

表皮生长因子受体(EGFR)家族包括EGFR、c-erbB-2、c-erbB-3和c-erbB-4,参与细胞转化和肿瘤发生。为了解葡萄胎和绒癌细胞中EGFR的表达强度及临床意义,对妊娠滋养细胞疾病和正常胎盘中EGFR、c-erbB-3、c-erbB-4的表达进行检测。     

胚胎干细胞因子Nanog、Oct4、Sox2与多囊卵巢综合征的相关性

目的探讨颗粒细胞中胚胎干细胞因子Nanog、Oct4、Sox2与多囊卵巢综合征(PCOS)发生的关系。方法选择接受体外受精/卵胞质内单精子注射-胚胎移植术(IVF/ICSI-ET)治疗的PCOS患者40例,同期输卵管因素或男方因素不孕患者40例作为对照组。利用Real time-PCR技术检测颗粒细胞中Nanog、Oct4、Sox2以及抗苗勒管激素(AMH)mRNA的表达水平,并用Western blotting分析蛋白表达水平与PCOS发生的关系。结果 PCOS组体质量指数(BMI)[(26.44±3.46)kg/m~2]、黄体生成素(LH)[(7.56±0.98)IU/L]、LH/卵泡刺激素(FSH)(1.06±0.15)及获卵数(28.7±3.9)较对照组[(22.43±3.17)kg/m~2、(4.26±0.34)IU/L、0.57±0.04、15.1±2.0]有显著升高(P均0.05)。Real time-PCR结果显示,PCOS组胚胎干细胞因子Nanog(0.60±0.09)和Oct4(0.85±0.14)mRNA表达水平显著低于对照组(1.65±0.17,1.59±0.17)(P0.05),AMH转录水平(1.28±0.11)显著高于对照组(0.89±0.07)(P0.05)。Logistic回归分析结果显示,Nanog是PCOS发生的危险因素(OR=26.577,P=0.002),且Nanog与BMI、获卵数及AMH呈显著负相关(r1=-0.415,P=0.023;r2=-0.415,P=0.022;r3=-0.373,P=0.043),与Oct4呈显著正相关(r=0.684,P0.01)。Western blotting结果显示,与对照组比较,PCOS组Nanog蛋白表达降低,AMH蛋白表达水平增高。结论 PCOS卵巢颗粒细胞中胚胎干细胞因子Nanog以及AMH的异常表达可能参与了PCOS的发生并影响卵母细胞的发育与成熟,为PCOS的临床治疗提供理论依据。     

E-钙粘素和β-连环素在妊娠滋养细胞疾病中的表达与意义

目的探讨E钙粘素和β连环素表达与妊娠滋养细胞疾病(GTD)恶性转化的相关性。方法应用逆转录聚合酶链反应技术检测12例正常早孕绒毛和35例妊娠滋养细胞疾病组织中E钙粘素mRNA和β连环素mRNA的表达。结果E钙粘素和β连环素mRNA在正常早孕绒毛与葡萄胎组织的表达差异无显著性(P>0.05);而在正常早孕绒毛、葡萄胎组织的表达量明显高于侵蚀性葡萄胎、绒癌组织(P<0.05);在侵蚀性葡萄胎组织的表达高于绒癌组织(P<0.05)。结论E钙粘素和β连环素的表达下调与妊娠滋养细胞疾病的恶性转化有关,对预测妊娠滋养细胞疾病的恶变以及评价预后有一定的参考价值。     

上皮型钙粘附素在妊娠滋养细胞疾病中的表达及与滋养细胞肿瘤侵袭的关系

目的 :明确上皮型钙粘附素 (E cd)在妊娠滋养细胞疾病中的表达及其与恶性滋养细胞肿瘤侵袭的关系。方法 :应用免疫组织化学SP方法检测葡萄胎、侵蚀性葡萄胎及绒毛膜癌石蜡切片中E cd的表达情况 ,并分析其与恶性滋养细胞肿瘤侵袭的关系。结果 :①E cd在葡萄胎、侵蚀性葡萄胎及绒癌的细胞滋养细胞中表达存在差别 (P <0 .0 0 1)。②E cd在侵蚀性葡萄胎及绒毛膜癌中细胞滋养细胞上的表达与葡萄胎比较均明显下降 (P <0 .0 0 1)。③侵蚀性葡萄胎及绒癌浸润子宫肌层的深度与E cd的表达状况差异有显著性 (P <0 .0 5 )。结论 :E cd表达下降或缺失与恶性滋养细胞的高侵袭力密切相关。     

子痫前期PI3K/Akt通路相关非编码RNA的研究进展

子痫前期（pre-eclampsia）是妊娠期常见并发症之一,是导致孕产妇和围生儿死亡的重要原因。子痫前期的发生与胎盘滋养细胞功能障碍密切相关。胎盘滋养细胞侵袭不全和子宫螺旋动脉重铸异常是子痫前期发生的主要病理机制。磷脂酰肌醇3激酶/蛋白激酶B(phosphatidylinositol-3 kinase/protein kinase B,PI3K/Akt)通路对胎盘滋养细胞的增殖、存活、凋亡、迁移和侵袭等过程具有重要调节作用。非编码RNA(non-coding RNA,ncRNA)中的微小RNA、长链非编码RNA和环状RNA具有重要的多水平基因调控功能,在PI3K/Akt通路中发挥重要调节作用,影响胎盘滋养细胞的功能,与子痫前期的发生、发展相关。综述近年子痫前期PI3K/Akt通路相关ncRNA的研究进展,为探索和发现子痫前期发病机制中的关键分子提供参考。     

层粘蛋白及受体在正常妊娠及妊娠滋养细胞疾病中的表达

目的探讨层粘蛋白(LN)及其受体(LN-R)在正常妊娠及妊娠滋养细胞疾病中的表达及其与恶性滋养细胞肿瘤侵袭的关系。方法收集1991年1月至2003年12月温州医学院附属一院正常妊娠、葡萄胎、侵蚀性葡萄胎及绒毛膜癌(绒癌)石蜡包埋组织,应用免疫组织化学SP方法检测各组中LN、LN-R的表达情况。结果LN、LN-R在正常妊娠、葡萄胎、侵蚀性葡萄胎及绒癌中的表达差异有非常显著性意义(P<0·01)。LN、LN-R在侵蚀性葡萄胎及绒癌中的表达与正常妊娠、葡萄胎比较差异有非常显著性意义(P<0·01)。结论LN、LN-R在妊娠滋养细胞疾病中的表达异常与恶性滋养细胞的高侵袭力密切相关。     

血管生成素在妊娠滋养细胞疾病中表达的临床意义探讨

目的:通过检测血管生成素(ANG)在人正常早孕绒毛组织、葡萄胎及妊娠滋养细胞肿瘤中表达的差异性,探讨ANG在妊娠滋养细胞疾病(GTD)表达的意义。方法:采用免疫组化(En Vision)法检测人正常早孕绒毛组织28例、葡萄胎组织26例、妊娠滋养细胞肿瘤组织14例(其中侵蚀性葡萄胎1例、绒毛膜癌12例、胎盘部位滋养细胞肿瘤1例)中ANG蛋白的表达以及定位情况。结果:ANG蛋白在人正常早孕绒毛组织、葡萄胎及妊娠滋养细胞肿瘤组织中均有表达且主要定位于细胞质和细胞膜、少量表达于细胞核;ANG阳性表达率葡萄胎组(100.0%)高于正常早孕绒毛组(60.7%)(P0.05)和妊娠滋养细胞肿瘤组(85.7%)(P0.05),妊娠滋养细胞肿瘤组高于正常早孕绒毛组(P0.05)。在妊娠滋养细胞肿瘤中,ANG阳性表达率在年龄≥40岁组(83.3%)高于年龄40岁组(75.0%)(P0.05);临床分期晚期组(Ⅲ、Ⅳ期)(88.9%)高于早期组(Ⅰ期、Ⅱ期)(80.0%)(P0.05);FIGO预后评分高危组(87.5%)高于低危组(83.3%)(P0.05)。结论:ANG在GTD组织中的表达明显高于在正常早孕绒毛组织,且在妊娠滋养细胞肿瘤晚期、高危患者表达增高,表明ANG的过表达可能参与了GTD的发生过程。     

Dysregulated activation of c-Src in gestational trophoblastic disease contributes to its aggressive progression

Introduction

Gestational trophoblastic disease (GTD) is a heterogeneous group of pregnancy-related disorders. Hydatidiform mole (HM) is the most common type of GTD, whereas gestational choriocarcinoma is the most aggressive. Non-receptor tyrosine kinase c-Src contributes to the transformation to a malignant phenotype in various cancers. However, the role of c-Src in the pathogenesis of GTD remains largely unknown.

Methods

The expression level of phosphorylated c-Src was determined by immunohistochemistry and Western blotting assay. JAR and JEG-3 cells were treated with hCG, specific c-Src inhibitor saracatinib and PP2, and PKA specific inhibitor, PKI. Cell growth rate and cell migration/invasion ability was determined by cell proliferation and transwell assays respectively.

Results

c-Src was highly activated in HM tissues and choriocarcinoma cells (JAR and JEG-3). c-Src was activated by hCG in a time and concentration-dependent manner, which was abrogated by specific c-Src and PKA inhibitors. Inhibition of c-Src activity in JAR and JEG-3 cells by saracatinib leaded to a decrease in the rate of cell growth and cell migration/invasion ability. Furthermore, inhibition of c-Src phosphorylation induced cell cycle arrest and reduced expressions of cyclin A2, cyclin B1, cyclin E1, FOXD3 and NANOG. Moreover, inhibition of c-Src activity resulted in decreased p-FAK^Tyr397 phosphorylation.

Discussion and conclusion

Our findings indicate an important role of c-Src in the pathogenesis of GTD, and we propose that c-Src inhibitors are potential adjuvant chemotherapeutic drugs for the treatment of GTD.     

妊娠滋养细胞疾病Cyclin B1、PCNA表达的研究

目的 :检测CyclinB1和增殖细胞核抗原 (PCNA)蛋白在妊娠滋养细胞疾病(GTD)中的表达 ,探讨二者的相关性及与GTD的关系。方法 :采用免疫组化S P法检测 1 5例正常绒毛、38例葡萄胎 (HM)、42例侵蚀性葡萄胎 (IM)和 1 8例绒毛膜癌 (CC)组织中Cy clinB1和PCNA蛋白表达情况。结果 :葡萄胎、IM、CC组织中的CyclinB1和PCNA蛋白表达水平显著高于正常绒毛。术前未化疗的IM和CC组织中CyclinB1的表达水平明显高于葡萄胎。CC组织PCNA的表达水平高于葡萄胎 (P =0 .0 0 5)。葡萄胎恶变者CyclinB1和PCNA的表达显著高于未恶变者。术前未化疗的滋养细胞肿瘤患者 (包括IM和CC)Cy clinB1表达显著高于术前化疗≥3疗程者 (P =0 .0 0 2 )。WHO预后评分为高危者及中危者的CyclinB1表达显著高于低危者 (P =0 .0 0 5)。PCNA的表达与滋养细胞肿瘤是否化疗、WHO分期、WHO评分无关。PCNA与CyclinB1的表达呈正相关 (P =0 .0 0 0 )。结论 :GTD中存在CyclinB1的调节紊乱 ,CyclinB1异常表达可能与葡萄胎滋养细胞的增生、恶变及滋养细胞肿瘤的恶性生物学行为有关     

Cytokeratin 20 as a biomarker of gestational trophoblastic disease: diagnostic and prognostic significance

OBJECTIVES: This study was designed to examine whether cytokeratin 20 (CK20) is expressed in molar pregnancies and may therefore be used in the diagnosis of gestational trophoblastic disease (GTD). The potential of CK20 expression in predicting the evolution and the prognosis of the different subtypes of GTD was also assessed. METHODS: A total of 48 samples were studied for CK20 expression by RT-PCR methodology. Among these, 24 samples were obtained by curettage of the uterine cavity of patients diagnosed with hydatidiform mole (14 complete moles and 10 partial moles), 4 samples were obtained from choriocarcinoma cell lines (2 JAR and 2 JEG), and 20 samples were of normal trophoblast (control group) obtained from patients that underwent elective termination of pregnancy. RESULTS: Expression of CK20 was identified in all the samples of complete mole (CM), all choriocarcinoma cell lines, and 50% of the patients with partial mole (PM). None of the preparations of normal trophoblastic tissue from the control group expressed the CK20. A significant difference (P < 0.00001) was found in CK20 expression between samples of patients with GTD and control samples. Comparison between CK20 expression in CMs and PMs revealed a significantly more frequent expression of CK20 in CMs (P = 0.006). More than 50% of the patients with PMs that were positive for CK20 had an invasive evolution. CONCLUSIONS: In our opinion, CK20 may assist in distinguishing between molar and normal trophoblastic tissue and may be considered a marker of GTD. In cases in which pathological classification of different subtypes of GTD is in doubt, CK20-positive expression is suggestive for a CM whereas CK20-negative is more indicative for PM.     

17β-雌二醇对子宫内膜癌细胞磷脂酰肌醇3激酶/蛋白激酶B信号传导通路的激活作用

目的　探讨 17β 雌二醇 (E2 )对子宫内膜癌细胞系Ishikawa细胞磷脂酰肌醇 3激酶 /蛋白激酶B(PI 3K/PKB)信号传导通路的激活作用以及PI 3K抑制剂对其的影响。方法　应用免疫印迹杂交技术 ,检测一定浓度 (1× 10 -6mol/L) 17β E2 作用于Ishikawa细胞不同时间 (分别为 0、5、15、30、6 0、12 0min)后和不同浓度 (分别为 0、1× 10 -10 、1× 10 -8、1× 10 -6、1× 10 -4mol/L) 17β E2 作用Ishikawa细胞一定时间 (30min)后PKB的活化情况 [以磷酸化PKB和总PKB比值 (p PKB/PKB)表示 ],以及用同法检测PI 3K抑制剂LY2 94 0 0 2对 17β E2 活化PKB的影响。结果　 1× 10 -6mol/L 17β E2 作用于Ishikawa细胞 15min时 ,PKB的活化水平 (0 5 33± 0 0 2 9)已明显高于对照 (0 36 1± 0 0 2 9,P <0 0 5 ) ,30min时最明显 (0 6 6 6± 0 0 2 1,P <0 0 0 1) ,而且持续至少达 12 0min时 ;随着 17β E2 浓度 (分别为 1× 10 -10 、1×10 -8、1× 10 -6、1× 10 -4mol/L)的增加 ,PKB的活化水平逐渐增高 ,与对照比较 ,差异有显著性 (除 1×10 -10 mol/L外 ,P值分别为 >0 0 5、<0 0 5、<0 0 5、<0 0 0 1)。随着LY2 94 0 0 2作用浓度 (分别为 0 1、10、5 0、10 0 μmol/L)的增加 ,17β E2 作用Ishikawa细胞后P     

hCG Secretion in human choriocarcinoma JAR cells is MAPK but not Stat3 dependent: contributions of TNFalpha and IL-1beta to inflammation-induced hCG secretion

Inflammatory mediators play critical physiologic roles throughout the course of normal pregnancy. At the same time, uncontrolled inflammation appears to have an adverse affect on pregnancy outcome. Thus, a crucial task of reproductive immunology is to define the mechanisms through which inflammation is controlled at the maternal-fetal interface. We examined the signaling pathways activated in the human choriocarcinoma cell line, JAR, in response to an in vitro model of an inflammatory challenge. We incubated JAR cells with medium from peripheral blood mononuclear cells (PBMC) that had been activated with either LPS or PHA. Conditioned medium from each experimental model induced MAP kinase and Stat3 phosphorylation as well as human chorionic gonadotropin (hCG) secretion from JAR cells. hCG secretion could be blocked by pharmacologic inhibition of MAP kinase but not by inhibition of Stat3 using an siRNA approach. The MAPK activators IL-1beta and TNFalpha both induced hCG secretion from JAR cells, but not the Stat3 activators IFN-gamma and IL-6. Furthermore, hCG secretion induced by conditioned media could be blocked by IL-1 receptor antagonist. We conclude that inflammation at the maternal-fetal interface may involve complex webs of regulatory and counter-regulatory mechanisms that involve multiple cytokines and at least two major, independent cell-signaling systems.     

瘦素、胰岛素及IGF-2影响滋养层细胞中PKB和P70S6K蛋白表达的机制

目的:探讨瘦素、胰岛素及IGF-2对滋养层细胞中PKB和P70S6K蛋白表达的影响是否与PI3K/PKB/mTOR信号途径存在联系。方法:应用Western blot检测瘦素、胰岛素及IGF-2作用后滋养层细胞中PKB和P70S6K蛋白的表达,以及分别加入PI3K的特异性抑制剂LY294002和mTOR的特异性抑制剂雷帕霉素后对PKB和P70S6K蛋白表达的影响。结果:瘦素、胰岛素及IGF-2刺激组滋养层细胞中PKB和P70S6K蛋白表达与对照组相比明显增加(P0.05)。瘦素、胰岛素及IGF-2刺激组分别加入LY294002和雷帕霉素,PKB和P70S6K蛋白表达均明显降低(P0.05)。结论:瘦素、胰岛素及IGF-2可能通过PI3K/PKB/mTOR信号通路促进滋养层细胞中PKB和P70S6K蛋白表达。