P53与妊娠滋养细胞肿瘤的增殖和凋亡

目的探讨P53在妊娠滋养细胞肿瘤(GTT)中的表达意义及其与GTT中细胞增殖、凋亡的关系。方法采用免疫组化SP法及核酸原位末端标记(TUNEL)法,检测妊娠滋养细胞肿瘤中P53和PCNA的表达率及细胞凋亡指数(AI)。结果在正常早期绒毛(NP)、完全性葡萄胎(CM)、侵袭性葡萄胎(IM)、绒癌(CCA)中,P53表达率(P53-Ⅰ)分别为4.12%、21.68%、39.61%和27.39%;PCNA表达率(PI)分别为10.40%、20.76%、53.60%和51.95%;凋亡指数(AI)分别为1.88%、2.59%、6.45%和1.26%;各组间比较均P<0.001。P53-Ⅰ和PI呈显著正相关(r=0.587,P<0.001),而与AI无明确的相关性。结论GTT的发生可能是由于机体的细胞增殖与凋亡调节系统失调所致;P53可能促进GTT中的细胞增殖活性,与GTT的发生、发展密切相关;未发现P53与GTT细胞凋亡的相关性。     

PCNA和Caspase-3在妊娠滋养细胞疾病中的表达及意义

目的研究PCNA和Caspase-3在妊娠滋养细胞疾病中的表达变化及意义,为妊娠滋养细胞疾病早期诊断、预后判断提供理论依据。方法收集滋养细胞疾病标本50例,其中葡萄胎10例(均为完全性葡萄胎),侵蚀性葡萄胎20例,绒毛膜癌20例。正常早孕绒毛标本10例为对照组。应用免疫组织化学技术检测PCNA和Caspase-3在早孕绒毛、葡萄胎、侵蚀性葡萄胎、绒毛膜癌组织中的表达情况。结果 PCNA在正常早孕绒毛、葡萄胎、侵蚀性葡萄胎和绒癌中的表达分别为10.24%、20.70%、50.92%、53.65%,各组差异有统计学意义(P0.05)。Caspase-3在正常早孕绒毛、葡萄胎、侵蚀性葡萄胎和绒癌中的表达先升高又降低,分别为1.25%、2.60%、6.40%、1.90%,各组差异有统计学意义(P0.05)。结论 PCNA及Caspase-3表达异常,增殖/凋亡失衡是引起GTT的重要因素。     

米非司酮对早孕绒毛滋养层细胞Bcl-2和caspase-3表达的影响

探讨米非司酮对人早孕绒毛滋养细胞中Bcl—2和caspase-3表达的影响及其在滋养细胞凋亡中的作用。方法：将105例自愿要求终止妊娠的早孕妇女随机分成两组，即米非司酮流产组(55例)及人工流产组(50例)。采用免疫组化染色方法检测早孕妇女的绒毛组织中Bcl—2和caspase-3的表达。结果：米非司酮组与人流组绒毛滋养细胞中Bcl—2的表达率分别为54．6％和88．0％；caspase-3的表达率分别为89．1％和42．0％。米非司酮组与人流组相比较绒毛滋养层细胞Bcl—2呈低表达(P＜0．01)，caspase-3呈高表达(P＜0．01)。结论：Bcl—2和caspase-3在米非司酮诱导的早孕绒毛滋养层细胞凋亡的信号传导中起关键作用。     

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。     

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     

妊娠滋养细胞肿瘤组织中cyclin D1、Rb蛋白产物的表达

目的：探讨增殖相关基因cyclin D1、Rb在妊娠滋养细胞肿瘤中的表达变化。方法：采用免疫组化S－P法,检测20例葡萄胎、15例侵蚀性葡萄胎、15例绒毛膜癌组织中两种基因蛋白产物的表达。结果：在恶性滋养细胞肿瘤组织中,cyclinD1的阳性表达率随临床期别的增高呈递增趋势,而Rb呈递减趋势,二者在Ⅲ期的阳性表达率与Ⅰ期及Ⅱ期相比差异均具有显著性（P＜0．05）;化疗可降低cyclinD1在恶性滋养细胞肿瘤中的阳性表达,而对Rb无影响。结论：cyclin D1的过表达,Rb的缺失在妊娠滋养细胞肿瘤的发展中可能有重要的生物学意义。     

E-cadherin与乳腺癌分化程度及其淋巴结转移的关系研究

目的 探讨E-cadherin基因表达在乳腺癌侵袭和转移中的作用。方法 采用原位杂交技术检测30例正常乳腺组织和48例乳腺癌原发肿瘤及其淋巴结转移灶中E-cadherinmRNA表达情况。结果 乳腺癌原发肿瘤组织和淋巴结转移灶中，E-cadherin mRNA阴性表达率（分别为31．3％，46．7％）与正常乳腺组织之间的差异非常显著（P＜0．001）。E-cadherin mRNA表达强度在乳腺癌Ⅰ-Ⅲ级之间及有无淋巴结转移的原发肿瘤之间有显著性差异（P＜0．05）。结论 肿瘤抑制基因E-cadherin是一种有价值的反映乳腺癌恶性程度和预测淋巴结转移的指标。     

Ki67在正常妊娠和胎儿生长受限患者胎盘组织中的表达和意义

目的 通过检测增殖蛋白Ki67在正常妊娠各期和FGR患者胎盘组织中的表达，探讨其在正常妊娠中的作用及与FGR的关系。方法 采取免疫组化法取正常妊娠各期和FGR患者胎盘组织共42例，检测Ki67蛋白的表达。结果 （1）Ki67主要在细胞滋养细胞中表达强，在合体滋养细胞中呈阴性。（2）Ki67在早、中、晚期胎盘组织阳性表达率无统计意义（P〉0．05），表达强度早、中期及中、晚期无显著差别（P〉0．05）；早、晚期有显著差异（P〈0．05）：FGR组与正常妊娠晚期组胎盘中Ki67表达阳性率、表达强度之间无显著差异（P〉0．05）。结论 （1）Ki67在妊娠早期、中期表达强度高，说明滋养细胞处于高增殖状态。（2）Ki67随妊娠进展表达下调，提示与胎盘老化和分娩发动有关。（3）FGR组Ki67表达阳性率上调说明细胞滋养细胞的增生。     

PCNA和P57^kip2在葡萄胎中表达的研究

目的探讨PCNA（增殖细胞核抗原,proliferation cell nuclear antigen）和P57^kip2在CM（完全性葡萄胎,com-plete hydatidiform mole）及PM（部分性葡萄胎,partial hydatidiform mole）中的表达及意义。方法应用免疫组化的方法检测PCNA及P57^kip2蛋白在CM、PM及HA（水肿绒毛,hydrops trophonema）组织中的表达。结果 PCNA在所有组织切片中均见到。PCNA主要在细胞滋养层细胞及中间型滋养细胞表达。PI在HA中最低,于PM中增高,CM中达到最高。其中CM与PM及与HA之间,差异均有统计学意义（P〈0.05）。而PM与HA之间无显著性差异（P〉0.05）。P57^kip2在30例HA27例呈阳性表达（90%）,30例PM 25例呈阳性表达（83.33%）,28例CM 3例呈阳性表达（10.71%）,并且在CM中的阳性强度均较弱。3种病变P57^kip2染色阳性率统计分析显示CM与PM及HA之间均存在显著性差异（P〈0.01）,而PM与HA之间无显著性差异（P〉0.05）。结论 PCNA与P57^kip2能代表葡萄胎滋养细胞异常增生的客观指标,P57^kip2蛋白的低表达和PCNA的高表达可能在葡萄胎的的发生发展中起重要作用,两者的共同检测对早期CM的发现有重大作用。     

JAM-A通过调控FAK/ERK信号通路对宫颈癌细胞进展的影响

目的 探讨连接黏附分子A(JAM-A)在宫颈癌细胞进展中的作用及其潜在分子机制。方法 实时荧光定量PCR(RT-qPCR)实验检测JAM-A在宫颈癌患者癌组织中的表达水平。将Vector、pcDNA-JAM-A、NC-siRNA、JAM-A-siRNA质粒分别转染至宫颈癌细胞中，CCK-8检测细胞增殖能力；Transwell实验检测细胞侵袭能力；细胞划痕实验检测细胞迁移能力；Western blotting分别检测细胞中JAM-A、PCNA、MMP-2、E-cadherin及FAK/ERK通路蛋白的水平。进行裸鼠皮下成瘤实验，检测JAM-A对宫颈癌细胞体内肿瘤生长的影响。结果 与癌旁组织比较，宫颈癌组织中JAM-A mRNA和蛋白水平均显著上调。过表达JAM-A可显著增强细胞增殖、侵袭和迁移能力，升高JAM-A、PCNA、MMP-2蛋白表达，降低E-cadherin蛋白表达。敲低JAM-A可显著抑制细胞增殖、侵袭和迁移能力，降低JAM-A、PCNA、MMP-2蛋白表达，升高E-cadherin蛋白表达。pcDNA-JAM-A+PF-562271组细胞中p-ERK2蛋白表达较pcDNA-...     

Minichromosome maintenance protein 7 expression in gestational trophoblastic disease: correlation with Ki67, PCNA and clinicopathological parameters

AIMS: To assess the proliferative activity of gestational trophoblastic disease (GTD) using one of the novel proliferation markers (MCM7) and to determine its prognostic value in hydatidiform mole (HM). METHODS AND RESULTS: Immunohistochemical staining for MCM7 was performed on 122 samples of paraffin-embedded trophoblastic tissues including 22 normal first-trimester placentas, 12 term placentas, 12 spontaneous miscarriages (SM), 21 partial moles (PM), 44 complete hydatidiform moles (CM), and 11 choriocarcinomas (CCA). The correlations between the proliferative indices assessed by MCM7, proliferating cell nuclear antigen (PCNA) and Ki67 (MIB1) immunoreactivity as well as clinical progress were assessed. MCM7 immunoreactivity was found predominantly in the nuclei of cytotrophoblast and intermediate trophoblast and decreased with placental maturation. MCM7 expression was highest in CCA, followed by CM, PM, normal first-trimester placenta, SM and term placenta. MCM7 index was significantly higher in PM and CM than in SM (P = 0.007, P < 0.001) but not between PM and CM themselves (P = 0.560). Eighteen of the 65 patients with HM developed persistent trophoblastic disease (PTD) requiring chemotherapy. There was no significant difference in MCM7 indices between the patients who developed PTD and those who did not (P = 0.312). MCM7 indices correlated well with Ki67 (P = 0.002) but not with PCNA (P = 0.054) indices. MCM7 indices demonstrated less variability than PCNA and Ki67 and may be a better proliferation marker than the latter two. CONCLUSIONS: We conclude that MCM7 is useful in differentiating molar and non-molar gestations but is not helpful in discriminating PM from CM or in predicting PTD.     

滋养细胞肿瘤DNA图像定量分析

目的:探讨图像分析技术对滋养细胞肿瘤DNA定量分析的应用价值.方法:应用图像分析技术对33例滋养细胞肿瘤细胞核DNA含量进行了定量测定和比较.结果:从正常绒毛→良性葡萄胎→恶性葡萄胎→绒毛膜癌:DNA含量和非整倍体细胞呈增高趋势.恶性葡萄胎和绒毛膜癌明显高于良性葡萄胎和正常绒毛,有显著性差异(P<0.01).良性葡萄胎高于正常绒毛,但两者相比无显著性差异(P>0.05).绒毛膜癌高于恶性葡萄胎,但两者相比亦无显著性差异(P>0.05).结论:图像分析仪对滋养细胞肿瘤DNA定量分析,可对该肿瘤的诊断、化疗、预后提供重要信息.     

p57和p53蛋白在水肿性流产及葡萄胎鉴别诊断中的作用

目的探讨p57和p53蛋白在水肿性流产、完全性和部分性葡萄胎鉴别诊断中的作用。方法分别收集正常绒毛、水肿性流产、部分性葡萄胎和完全性葡萄胎石蜡标本10、12、23和20例,应用免疫组织化学EnVision法检测p57和p53蛋白在这些组织中的分布及表达水平。结果p57蛋白在正常绒毛、水肿性流产及部分性葡萄胎组织中主要分布于绒毛的细胞滋养细胞及间质细胞,阳性表达比例分别为10/10、12/12和100％( 23/23),各组间相比差异无统计学意义(P＞0.05)。在完全性葡萄胎中细胞滋养细胞及间质细胞p57表达缺失,与部分性葡萄胎相比,差异有统计学意义(P＜0.05)。p53蛋白主要表达于完全性和部分性葡萄胎的细胞滋养细胞及中间滋养细胞。在正常绒毛中p53蛋白呈阴性表达,水肿性流产中仅1例p53蛋白呈阳性表达(1/12),部分性葡萄胎和完全性葡萄胎中p53蛋白的阳性率分别为60.9％( 14/23)和85.0％ (17/20);p53蛋白的阳性率,部分性葡萄胎较水肿性流产明显增加,完全性葡萄胎较部分性葡萄胎也明显增加,差异均有统计学意义(均P ＜0.05)。结论p57蛋白免疫组织化学检测可辅助鉴别完全性和部分性葡萄胎,而p53蛋白的检测则有助于鉴别水肿性流产和部分性葡萄胎。     

Subsequent pregnancy outcome in patients with spontaneous resolution of HCG after evacuation of hydatidiform mole: comparison between complete and partial mole

This study compared subsequent pregnancy outcome in patients with complete and partial hydatidiform moles. Among 1052 patients with molar pregnancy (complete mole, 801; partial mole, 251) monitored at Chiba University Hospital between 1981 and 1999, 891 patients (84.7%) had spontaneous resolution of human chorionic gonadotrophin (HCG) after mole evacuation, and 161 patients (15.3%) required chemotherapy. Of the 891 patients, 438 (49.2%) had 650 subsequent pregnancies. The pregnancy outcome was not significantly different in patients with complete and partial moles, and was comparable with that in the general Japanese population. The incidence of repeat molar pregnancy in patients with complete and partial mole (1.3 and 1.5% respectively) was 5-fold higher than that of the general population, while no increased risk of persistent gestational trophoblastic tumour (GTT) associated with later molar pregnancy was observed. During HCG follow-up, 10 patients (1.1%) developed secondary high-risk GTT between 14 and 54 months after mole evacuation. The incidence of high-risk GTT in patients with and without subsequent pregnancies was 0.46% (2/438) and 1.8% (8/453) respectively (P = 0.1243). In conclusion, patients with complete and partial mole can anticipate a normal future reproductive outcome, and pregnancies after experiencing hydatidiform mole may not affect the development of high-risk GTT.     

Use of proliferation cell nuclear antigen immunoreactivity for distinguishing hydropic abortions from partial hydatidiform moles.

AIMS: To determine whether the expression of proliferating cell nuclear antigen (PCNA) in villous cytotrophoblast could distinguish between placental tissue from a hydropic abortion and that from a partial hydatidiform mole. METHODS: Tissue from 18 partial hydatidiform moles, 15 hydropic abortions, five normal first trimester placentas and five normal full term placentas were immunostained for expression of PCNA, using the monoclonal antibody PC10. RESULTS: PCNA immunoreactivity was very much higher in the cytotrophoblast of normal first trimester placentas than in normal term placentas. Villous tissue from partial hydatidiform moles showed, on average, less immunoreactivity for PCNA than did villous tissue from hydropic abortions. CONCLUSIONS: Immunostaining for PCNA is of no value for differentiating between partial hydatidiform moles and hydropic abortions. The findings indicate that trophoblastic proliferation or hyperplasia is not a feature of partial hydatidiform moles.     

Telomerase activity in gestational trophoblastic disease

AIMS: To investigate the pattern of telomerase activity in hydatidiform mole as compared with normal placenta and choriocarcinoma, and to determine the prognostic significance of telomerase activity in hydatidiform mole. METHODS: Telomerase activity in 35 cases of hydatidiform mole, 35 normal placentas, one choriocarcinoma sample, and two choriocarcinoma cell lines (JAR, JEG3) was determined using the sensitive polymerase chain reaction based telomeric repeat amplification protocol (TRAP) assay. Two cases of breast carcinoma and two cases of ovarian carcinoma were also included as positive controls in the telomerase assay. RESULTS: Telomerase activity was detected in 11 of 30 early placentas (36.7%), one of five term placentas (20%), five of 27 hydatidiform moles which regressed spontaneously (18.5%), and six of eight hydatidiform moles which developed persistent trophoblastic disease (75%) (including three which developed metastases). Hydatidiform moles which subsequently developed persistent disease, especially those which metastasised, were more likely to express telomerase activity (p < 0.01). However, there was no significant difference in the frequency of telomerase activity between early placentas and hydatidiform mole. Strong telomerase activity was observed in choriocarcinoma tissue, choriocarcinoma cell lines, and ovarian and breast carcinomas. CONCLUSIONS: Telomerase activation occurs in hydatidiform mole with a similar incidence to early normal placentas. This supports the concept that hydatidiform mole is essentially an abnormal conceptus. There is an association between telomerase activation and the development of persistent trophoblastic disease. Further study is warrant to confirm the prognostic significance of telomerase activity in hydatidiform mole.     

The significance of proliferating cell nuclear antigen in human trophobiastic disease: an immunohistochemical study

The expression of proliferating cell nuclear antigen (PCNA) in human trophobiastic disease was assessed immunohistochemically in tissue from 29 spontaneous abortions, 33 partial moles, 40 complete moles and 23 chorio carcinomas using the monoclonal antibody PC ID. PCNA immunoreactivity occurred predominantly in the cytotrophoblasts in each of the four types of tissues. Quantitative analysis showed that the choriocarcinoma group gave a statistically significant higher PCNA index than the other three. There was no significant difference between the groups of spontaneous abortion, partial or complete mole. Sixteen of the 73 patients with partial and complete moles developed persistent gestational trophobiastic disease and there was no significant difference between the patients requiring chemotherapy and those who did not. We conclude that choriocarcinoma has a significantly higher PCNA proliferative index whilst hydatidiform moles cannot be distinguished from abortions by such analysis. The PCNA index does not appear to be useful in predicting the progression of molar pregnancies to persistent trophobiastic diseases.