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。     

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     

葡萄胎中c-myc和ras癌基因表达及其意义

目的　研究探讨葡萄胎中c myc、ras基因表达情况及其意义。方法　采用免疫组织化学染色对 4 2例葡萄胎和10例早孕绒毛标本中c myc、ras基因表达进行检测 ,并采用图像分析技术 ,对正常早孕绒毛组和葡萄胎不同病理分级组间c myc、ras的表达情况进行对比分析。结果　c myc、ras阳性信号表达在正常早孕绒毛和葡萄胎Ⅰ Ⅲ级各组间均无明显统计学差异 (P >0 0 5 )。结论　葡萄胎中c myc、ras癌基因表达情况与病理学分级无关 ,与正常绒毛也无统计学差异 ,支持以下观点 :即大多数葡萄胎只是流产的一种形式而非真正的肿瘤。     

用单克隆抗体免疫测定和原位杂交技术确定人胃腺癌中c-Ha-rasP21的表达增强

ras癌基因族的成员包括有Ha-ras,Ki-ras及N-ras。迄今已有不少报道,证明它们为一系列种属,其中包括人的正常细胞成分。然而点突变可以引起ras基因质的改变,从而导致某些人类肿瘤的形成。另外若原-ras基因(proto-ras gene)表达水平增高也可使细胞发生恶性转化。本研究旨在探讨ras癌基因是否与人胃腺癌有关;rasP21的表达是否与肿瘤分化程度、组织学亚型(histological subtype)或肿瘤浸润深度有关。研究材料取自外科手术切除的或活检冰     

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。     

癌基因研究新发现与进展（二）

五.ras族癌基因 ras癌基因已成为ras族癌基因,除Ha-ras,Ki-ras,N-ras,还有酵母的Ras-1,Ras-2。ras样蛋白的特点为具有高度保守性,如人的c-Ha-ras蛋白(p21~(Ha—ras)),小鼠肉瘤病毒的Ha-ras蛋白,和酵母的Ha-ras蛋白的氨基酸顺序有36％相同。 ras癌基因在许多肿瘤中都已被检定,被认为它与信号传导关系密切,因为牵连到磷酸肌醇和鸟苷三磷酸活化蛋白(GAP)的体系,所以ras基因的研究除了解肿瘤的发病机制外,还有可能解决细胞     

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.     

MM P-2/TI MP-2及VEG F在滋养细胞疾病中的表达及意义

目的 检测不同滋养细胞疾病组织中MMP-2/TIMP-2及VEGF的表达情况,观察比较它们的表达特点,分析与滋养细胞疾病发生、发展的相关性.方法 收集30例正常绒毛;40例葡萄胎组织,30例手术切除的恶性葡萄胎组织,采用单克隆抗体免疫组化SP法对其染色,光镜下分别观察以上不同组织中MMP-2、TIMP-2、VEGF的阳性表达情况.结果 (1)从正常绒毛→良性葡萄胎→恶性葡萄胎,MMP-2的阳性率有升高趋势,差异有统计学意义(P＜0.05).VEGF的表达与MMP-2正相关.(2)TIMP-2的阳性表达在滋养细胞疾病内部不同分组间两两比较,差异均无统计学意义(P＞0.05).(3)MMP-2/TIMP-2组合同时阴性表达在良性葡萄胎中占65%(26/40),在恶性葡萄胎中占16.67%(5/30),两者比较有显著性差异(P＜0.01).MMP-2阳性表达伴TIMP-2阴性表达在良性葡萄胎中占2.5%(1/40),在恶性葡萄胎中占56.67%(17/30),两者比较有显著性差异(P＜0.01).结论 (1)MMP-2和VEGF共同参与了滋养细胞疾病的发生和发展.(2)MMP-2/TIMP-2两者联合检测对鉴别良、恶性滋养细胞疾病有重要价值.     

恶性滋养细胞肿瘤47例临床分析

目的总结恶性滋养细胞肿瘤的诊治经验,探讨恶性滋养细胞肿瘤的治疗方法及效果。方法回顾分析我院47例恶性滋养细胞肿瘤的临床资料。结果侵蚀性葡萄胎42例,绒癌5例。平均发病年龄31.5岁,47例治疗均以化疗为主,其中单纯化疗41例,化疗加手术治疗6例。侵蚀性葡萄胎治愈率97.6%（1/42）,绒癌治愈率100%（5/5）。结论临床上将病理、血HCG以及B超有机的结合起来使恶性滋养细胞肿瘤能够得到早期诊断,化疗对恶性滋养细胞有较好的疗效。     

层粘连蛋白和纤维粘连蛋白在正常绒毛膜葡萄胎、绒毛膜癌组织中的表达

目的探讨层粘连蛋白和纤维粘连蛋白在正常绒毛膜和滋养细胞肿瘤中的作用。方法用免疫组织化学方法观察层粘连蛋白和纤维粘连蛋白在正常绒毛膜、葡萄胎和绒毛膜癌组织中的表达。结果在正常绒毛膜组织中,二种蛋白有广泛表达,如绒毛上皮基膜、毛细血管内皮基膜、绒毛间质、滋养细胞柱和蜕膜。在葡萄胎,层粘连蛋白分布于绒毛上皮基膜和蜕膜组织,在基膜中表达强度有所不同;纤维粘连蛋白分布于绒毛和蜕膜内,呈弱阳性表达。在侵蚀性葡萄胎和绒毛膜癌,层粘连蛋白和纤维粘连蛋白在细胞质中均呈强阳性表达,在细胞间质内呈阳性或弱阳性表达。结论层粘连蛋白和纤维粘连蛋白与孕早期绒毛膜和蜕膜的形成及滋养细胞肿瘤的发生、侵润密切相关。     

A study of early pregnancy factor activity in the sera of women with trophoblastic tumor.

PROBLEM: To detect whether or not the early pregnancy factor (EPF)-like activity, or chaperonin 10, could be in the sera of patients with trophoblastic tumor in order to find another more efficient means to diagnose this kind of tumor. METHOD OF STUDY: The rosette inhibition assay was used to detect EPF-like activity in 216 sera, collected from patients with gestational trophoblastic tumor, including 47 sera of patients with choriocarcinoma, 68 sera of patients bearing invasive mole, and 101 sera of patients with vesicular mole. RESULTS: The accuracy of diagnosing malignant trophoblastic tumor by detecting EPF-like activity is 91.3% (105/115), with a false positive rate of 14.58% and a false negative rate of 4.8% by this method. Furthermore, the rosette inhibition titer (RIT) values have significant difference (P < 0.001) between the sera in patients with malignant trophoblastic tumor before treatment and those after treatment. CONCLUSIONS: This study demonstrated that diagnosis of malignant trophoblastic tumor could be made with an accuracy of 91.3% by detecting EPF-like activity and that EPF-like activity could be used as an indicator to distinguish benign from malignant trophoblastic tumor.     

Trophoblastic neoplasia in an African urban population.

A clinical study of trophoblastic neoplasia in a Nigerian population in Lagos over a four-year period is reported. A high incidence of one in 379 deliveries for benign trophoblastic tumor and one in 846 deliveries for malignant tumor was found. Seventeen percent of benign trophoblastic tumors in this series progressed to the malignant type, but malignant trophoblastic tumor was preceded by the benign type (hydatidiform mole) in 46 percent of cases. The anterior vaginal wall is a common site for metastases of malignant trophoblastic neoplasia and, in one patient, the lesion progressed further to form a vesicovaginal fistula. While the management of benign disease was conservative, all cases of malignant trophoblastic neoplasia received chemotherapy.     

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.     

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.     

Specific inhibitory substance to the husband's HLA allotype in sera from patients with gestational trophoblastic disease detected by lymphocytotoxicity testing

The sera of nine patients with gestational trophoblastic disease were investigated for the inhibitory substance specific to the HLA allotypes of the patients' husbands by microcytotoxicity inhibition testing using monoclonal antibodies (anti-A2, A9, A10) and HLA-typing serum to Bw52. The specific inhibitory substance was present in three of three choriocarcinoma patients, in three of four invasive mole patients, and in one of two hydatidiform mole patients and seemed to change in titer parallel with tumor growth. These facts suggest that the inhibitory substance could have some relation to the patient's immunological response against the tumor; that is, this inhibitory substance could be the reason why patients could not induce graft rejection response to trophoblastic tumor cells.     

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.