【作者】 张艳；

【导师】 闫晓娟；

【作者基本信息】 河北医科大学 ， 妇产科学， 2009， 硕士

【摘要】 目的:妊娠滋养细胞疾病(Gestational trophoblastic disease, GTD)是妊娠期特有的疾病,是一组来源于胎盘滋养细胞的疾病,包括葡萄胎、侵袭性葡萄胎、绒毛膜癌和胎盘部位滋养细胞肿瘤等。除葡萄胎外,又统称为妊娠滋养细胞肿瘤(Gestational trophoblastic tumor, GTT)。侵蚀性葡萄胎以及大约50%的绒毛膜癌都是由萄葡胎恶变而来。妊娠滋养细胞肿瘤恶性程度极高,极易侵蚀血管并发生远处转移,严重危害妇女生育功能及生命健康。葡萄胎恶变发生的确切原因,目前尚不完全清楚。因此,研究葡萄胎恶变机制,对于早期预测葡萄胎的恶变,指导临床预防性化疗,改善妊娠滋养细胞疾病的预后有重要意义。核转录因子-κB(Nuclear transcription factor-kappa B)NF-κB,是一种具有基因转录多项调控作用的细胞因子,在机体的免疫应答、炎症反应及细胞生长发育、凋亡等过程中发挥重要作用,并与多种肿瘤转移有密切关系。研究显示NF-κB存在于人类滋养细胞中,故提示NF-κB有可能在GTD的发病机制中起重要作用。研究表明金属基质蛋白酶-2(matrix metalloproteinase-2,MMP-2)、金属基质蛋白酶-9(matrix metalloproteinase-9,MMP-9)与肿瘤转移密切相关,因此本课题拟通过分析正常妊娠和葡萄胎组织中NF-κB和MMP-2、MMP-9的表达以及三者间的相关性,以探讨其在葡萄胎恶变中的可能的作用机制,从而为葡萄胎恶变的临床防治提供理论依据。方法:选取河北医科大学第三医院妇产科2000年-2006年住院葡萄胎病人第一次清宫术后葡萄胎组织标本,经病理学检查确诊为葡萄胎,共41例。其中包括经随访发现未发生恶变的葡萄胎29例称为葡萄胎未恶变组,恶变的葡萄胎12例,称为葡萄胎恶变组。所有病例均有完整的临床病理资料,其诊断标准参照宋鸿钊教授提出的妊娠滋养细胞疾病诊断标准。另外,选取正常相应妊娠绒毛组织17例作为对照组,通过免疫组化法检测正常妊娠和葡萄胎组织中NF-κB和MMP-2、MMP-9的表达,每个切片随机挑选5个视野,用HMIAS-2000图像处理系统进行定量灰度扫描,取平均灰度值。灰度值越低,则蛋白表达越多。应用SPSS 13.0统计软件进行数据处理。临床一般资料为计量资料采用单因素方差分析,P<0.05有统计学意义;相关性分析用Pearson积差相关,以α=0.05作为差异显著性水准。结果:1临床一般资料比较:各组间年龄、孕周的差异均无统计学意义(P>0.05)。2 NF-κB、MMP-2以及MMP-9在组织中的表达及定位:活化的NF-κB蛋白主要表达于绒毛滋养细胞和血管内皮细胞,定位在细胞浆,偶见于细胞核;MMP-2表达位于合体滋养细胞及绒毛外滋养细胞,定位在胞膜及胞浆中;MMP-9主要表达于细胞滋养细胞,定位在胞膜及胞浆。3 NF-κB的阳性表达在对照组、葡萄胎未恶变组以及葡萄胎恶变组的灰度值分别为137.82±6.66;115.55±9.61及97.15±11.15,F=71.22,P<0.01,各组间两两比较P<0.05,说明NF-κB在对照组、葡萄胎未恶变组和葡萄胎恶变组的表达均存在显著性差异。灰度值越低,则蛋白表达量越多。葡萄胎恶变组NF-κB的表达量明显高于未恶变组及正常绒毛,葡萄胎未恶变组NF-κB的表达量亦高于正常绒毛组的表达。与葡萄胎未恶变组比较,在葡萄胎恶变组组织中NF-κB核表达率明显升高(P<0.05),核表达率增加20%,有显著性差异。4 MMP-2的阳性表达在对照组、葡萄胎未恶变组及葡萄胎恶变组灰度值分别为105.64±5.30;102.40±7.13及70.93±8.87,F=103.21,P<0.01,灰度值越低,则蛋白表达量越多。葡萄胎恶变组MMP-2的阳性表达明显高于未恶变组及对照组(P<0.05),但未恶变组与对照组相比无显著性差异(P>0.05)。5 MMP-9的阳性表达在对照组、葡萄胎未恶变组及葡萄胎恶变组的灰度值分别为111.27±4.33;109.38±5.60及86.91±8.81,F=69.66,P<0.01,灰度值越低,则蛋白表达越多。葡萄胎恶变组MMP-9的阳性表达明显高于未恶变组及对照组(P<0.05),但未恶变组与对照组相比无显著性差异(P>0.05)。6葡萄胎组织中NF-κB与MMP-2及MMP-9的相关性:NF-κB在葡萄胎恶变组的表达与MMP-2和MMP-9分别呈正相关(r=0.890,P<0.01;r=0.713,P<0.01)。结论:1在葡萄胎发生恶变的组织中NF-κB、MMP-2及MMP-9的表达明显高于正常绒毛组织以及葡萄胎未发生恶变的组织。2葡萄胎恶变组中NF-κB与MMP-2及MMP-9表达呈正相关,且NF-κB核表达率明显升高,提示葡萄胎组织中MMP-2及MMP-9的表达受NF-κB的调控。3推测葡萄胎的预后,NF-κB、MMP-2及MMP-9蛋白表达的强弱与葡萄胎的预后有关,葡萄胎组织中NF-κB、MMP-2及MMP-9表达增高者预后差,侵蚀性葡萄胎或绒癌发生率高;反之,葡萄胎组织中NF-κB、MMP-2及MMP-9低表达者预后较好,侵蚀性葡萄胎或绒癌发生率低。4 NF-κB、MMP-2及MMP-9参与了葡萄胎恶变,对葡萄胎的预后有预测作用。更多还原

【Abstract】 Objective: Gestational trophoblastic diseases (GTDs) are originated from embryo trophoblast,and they are pregnancy associated diseases, including hydatidiform mole, invasive mole, choriocarcinoma and placental site trophoblastic tumor. The last three are termed gestational trophoblastic tumor (GTT). All invasive mole and around 50% of choriocarcinoma are derived from hydatiform mole. Invasive mole and choriocarcinoma are high malignant tumors characterized by blood vessels erodsion and distant metastasis. They are severly threatening womens’reproductive function and health. So far, it is still not clear about the etiology of malignant transformation of hydatidiform mole. Therefore, to study the mechanism of malignant transformation of hydatidiform mole will help us evaluate the prognosis of hydatidiform mole, provide the evidence for proper clinical treatment as well as reduce the incidence of malignant transformation of hydatidiform mole.Nuclear transcription factor-κB (NF-κB) is a gene transcription factor, which can regulate a wide range of cytokines in immune response, inflammatory response, cell growth and apoptosis, and it is also associated with a variety of tumor metastasis. Studies have shown that NF-κB exists in the human trophoblastic cells, suggesting that NF-κB may play an important role in the pathogenesis of GTD. It has been shown that matrix metalloproteinase-2(MMP-2),matrix metallo proteinase-9(MMP-9)are closely related to tumor metastasis. Therefore, we explored the possible roles of NF-κB, MMP-2 and MMP-9 in malignant transformation of hydatidiform mole by examining the expressions of NF-κB, MMP-2 and MMP-9, as well as their correlations. Our current study will provide a theoretical basis for clinical prevention and treatment of malignant transformation of hydatidiform mole.Methods: 41 cases molar tissues (including 29 cases molar tissues without maliganant transformation and 12 cases molar tissues with maliganant transformation through follow-up visiting) were collected from patients with hydatidiform mole who received initial evacuation in the Third Hospital of Hebei Medical University between 2000-2006, and 17 cases normal villi samples (normal control group) were derived from early pregnant women who received artificial abortion. All 41 cases with complete information and pathologic results were diagnosed according to the diagnostic criteria of trophoblatic diseases proposed by Song Hongzhao. The expressions of NF-κB, MMP-2 and MMP-9 were detected by immunohistochemical staining. Five areas on each section were randomly selected, and HMIAS-2000 image processing system was used to carry out Quantitative gray-scale scanning.The average gray value was used for statistic analysis. The lower the gray value, the higher the protein was expressed.The experimental results were analyzed by using SPSS 13.0 statistical software. General clinical data were analyzed by using one-way ANOVA, statistical significances were inferred at P<0.05; correlation analysis using Pearson product-moment correlation to setα= 0.05 as a standard of significant differenceResults:1 Comparison of general clinical information: The inter-group age and gestational age were not statistically significant (P> 0.05).2 The expressions and locations of NF-κB, MMP-2 and MMP-9 in tissues: The expression of activated NF-κB was mainly located in cytoplasm of trophoblastic cells and endothelial cells of blood vessels, occasionally found in the nucleus; MMP-2 expression was located in cell membrane and cytoplasm of syncytiotrophoblast and extravillous trophoblastic cells; MMP-9 expression was located in cell membrane and cytoplasm of cytotrophoblast cells3 The gray-scale values of NF-κB positive expressions were 137.82±6.66; 115.55±9.61 and 97.15±11.15, respectively in control group, hydatidiform mole without malignant transformation group and hydatidiform mole with malignant transformation group (F=71.22, P<0.01). The expressions of NF-κB were significantly different between each pair of groups (P<0.05). The lower the gray value, the more protein expressed. The expression of NF-κB in hydatidiform mole with malignant transformation group was much higher than those of control group and hydatidiform mole without malignant transformation group, in addition, the NF-κB expression in hydatidiform mole without malignant transformation group was also higher than that of normal control. The nuclear expression of NF-κB in hydatidiform mole with malignant transformation group was much higher than hydatidiform mole without malignant transformation group with 20% (P<0.05), there is significant difference.4 The gray-scale values of MMP-2 positive expressions were 105.64±5.30; 102.40±7.13 and 70.93±8.87, respectively in control group, hydatidiform mole without malignant transformation group and hydatidiform mole with malignant transformation group (F=103.21, P<0.01). The lower the gray value, the more protein expressed. The expression of MMP-2 in hydatidiform mole with malignant transformation group was much higher than those of control and hydatidiform mole without malignant transformation groups (P<0.05), while, there was no significant difference between hydatidiform mole without malignant transformation group and control group (P> 0.05).5 The gray-scale values of MMP-9 positive expressions were 111.27±4.33; 109.38±5.60 and 86.91±8.81, respectively in control group, hydatidiform mole without malignant transformation group and hydatidiform mole with malignant transformation group (F=69.66, P<0.01). The lower the gray value, the higher protein expression. The expression of MMP-9 in hydatidiform mole with malignant transformation group was much higher than those of control group and hydatidiform mole without malignant transformation group (P <0.05), while, there was no significant difference between hydatidiform mole without malignant transformation group and control group (P>0.05).6 The correlation of NF-κB with MMP-2 and MMP-9 in hydatidiform mole tissues: the expression of NF-κB in hydatidiform mole with malignant transformation group was positive correlated to MMP-2 and MMP-9, respectively (r = 0.890, P<0.01; r =0.713, P<0.01).Conclusion:1 The expressions of NF-κB, MMP-2 and MMP-9 in hydatidiform mole with malignant transformation group were significantly higher than those of control and hydatidiform mole without malignant transformation groups.2 The expression of NF-κB was positively correlated with both MMP-2 and MMP-9 in hydatidiform mole with malignant transformation group, and the nuclear expression of NF-κB was increased, suggesting that the expression of MMP-2 and MMP-9 were regulated by NF-κB.3 To evaluate the prognosis of hydatidiform mole: the expression extensity of NF-κB, MMP-2 and MMP-9 was associated with the prognosis of hydatidiform mole. The prognosis was worse in patients with higher expressions of NF-κB, MMP-2 and MMP-9 mole compared with those with lower expressed mole, and the incidence developing to invasive mole or choriocarcinoma was also higher in these patients. In contrast, the patients with lower levels of NF-κB, MMP-2 and MMP-9 had better prognosis and lower incidence of developing to invasive mole or choriocarcinoma.4 NF-κB, MMP-2 and MMP-9 are involved in malignant transformation of hydatidiform mole, and have an impact on trophoblastic invasion, suggesting that they play an important role in predicating the prognosis of hydatidiform mole.更多还原