【作者】 刘连连；

【导师】 马晓艳；

【作者基本信息】 吉林大学 ， 临床医学， 2011， 硕士

【摘要】 卵巢癌在妇科肿瘤中死亡率居首位。由于其早期没有特异的临床症状而不易被察觉,多数患者就诊时已属晚期,因此提高其早期诊断率尤为重要。转化生长因子-β(TGF-β)对正常细胞的生长起负调控作用,具有广泛的生物学活性,不仅调节细胞的增殖、分化、迁移和凋亡,还与细胞外基质的形成、免疫抑制及机会性感染等密切相关。TGF-β1是TGF-β的主要存在形式,活性最强。Smad4是TGF-β超家族信号传导通路的中心环节,50%胰腺癌中存在其突变、缺失。TGF-β1与Smad4在人类多种肿瘤的发生中起重要作用。本实验是通过检测卵巢肿瘤和正常对照组血清TGF-β1和Smad4水平及卵巢组织中Smad4的表达状况,为卵巢癌的发病机制、诊断、监测、治疗及预后提供一定的实验依据。材料和方法:采集卵巢癌、卵巢交界性肿瘤、卵巢良性上皮性肿瘤、卵巢性索间质肿瘤和正常对照组患者/妇女的空腹外周静脉血及组织,通过酶联免疫吸附实验检测其血清TGF-β1、Smad4水平,通过免疫组织化学检测Smad4在卵巢组织(上皮和间质)中的表达状况。结果:1、卵巢癌、卵巢交界性肿瘤及卵巢性索间质肿瘤患者术前血清TGF-β1水平差别无统计学意义(P>0.05),而三者均显著高于卵巢良性上皮性肿瘤及正常对照组患者血清TGF-β1水平,差别有统计学意义(P<0.01);卵巢良性上皮性肿瘤患者血清TGF-β1水平显著高于卵巢正常对照组,差别有统计学意义(P<0.01)。五组术后血清TGF-β1水平均显著高于其术前水平,差别有统计学意义(P<0.05),而术后各组之间差别无统计学意义(P>0.05)。卵巢癌患者术前血清TGF-β1水平与卵巢癌组织学分类及腹水量多少有关(P<0.05)。2、卵巢癌、卵巢交界性肿瘤、卵巢良性上皮性肿瘤及卵巢性索间质肿瘤患者术前血清Smad4水平高于正常对照组,差别有统计学意义(P<0.05)。卵巢癌患者术后血清Smad4水平低于卵巢良性上皮性肿瘤和卵巢性索间质肿瘤,差别有统计学意义(P<0.05)。卵巢良性上皮性肿瘤及正常对照组患者术后血清Smad4水平较术前升高,差别有统计学意义(P<0.05)。卵巢癌患者术前血清Smad4水平与组织学分级、组织学分类和腹水量有关(P<0.05)。3、Smad4蛋白在卵巢癌、卵巢交界性肿瘤、卵巢良性上皮性肿瘤、卵巢性索间质肿瘤和正常卵巢上皮组织中的缺失性表达率分别为46.3%(38/82)、40.0% (8/20)、58.3%(14/24)、25.0%(4/16)、12.5%(2/16)。其中,82例卵巢癌中有14例是粘液性腺癌(上皮缺失性表达率为85.7%,12/14),20例卵巢交界性肿瘤中12例是粘液性者(上皮缺失性表达率为50%,6/12),24例卵巢良性上皮性肿瘤中14例是粘液性囊腺瘤(上皮缺失性表达率为71.4%,10/14)。另外,有2例卵巢粘液性肿瘤组织中同时存在良性、交界性和恶性三种病变,Smad4在上述病变中均呈缺失性表达。Smad4蛋白在卵巢癌上皮组织中的缺失性表达率高于正常卵巢组织,差别有统计学意义(P〈0.05),卵巢良性上皮性肿瘤的上皮组织缺失表达率高于卵巢性索间质肿瘤和正常卵巢组织,差别有统计学意义(P〈0.05)。Smad4蛋白在卵巢癌间质中的缺失性表达率为24.4% (20/82),而其余各组卵巢间质中无缺失性表达。Smad4蛋白在卵巢癌上皮细胞中的缺失性表达率与患者年龄、组织学分类、临床分期、腹水量及淋巴结转移有关(P<0.05)。Smad4蛋白在卵巢癌间质细胞中的缺失性表达率与本文所有临床病理指标均无关(P>0.05)。结论:1、血清TGF-β1水平增高和Smad4水平增高均有可能作为诊断卵巢上皮性肿瘤和卵巢性索间质肿瘤的生物学指标。2、血清TGF-β1水平增高和Smad4水平增高均有可能成为判定卵巢癌预后不良的指标。3、卵巢上皮性肿瘤和卵巢性索间质肿瘤血清Smad4水平增高可能来源于卵巢肿瘤的上皮组织和间质组织。4、Smad4蛋白在卵巢癌组织中的缺失性表达可能是部分患者TGF-β1-Smad4信号传导通路障碍的重要原因之一,并有可能作为判定卵巢癌预后不良的指标之一。5、Smad4不能作为鉴别原发性和转移性卵巢粘液性癌的生物学指标。6、Smad4对于其在卵巢癌组织中缺失性表达的患者可能具有潜在的生物学治疗价值。更多还原

【Abstract】 Ovarian cancer has the highest mortality of all gynecological malignancies. Because the symptoms of early-stage ovarian cancer are not specific, it is not easy to detect. In the majority of patients, ovarian cancer is not detected until late stages. It is particularly important to improve the early diagnostic rate of ovarian cancer. Transforming growth factor-β(TGF-β)plays a negative role in normal cells growth regulation, it plays its biological effects in a broad range, not only the regulation of cell proliferation, differentiation, migration and apoptosis, but also the close relationship with extra cellular matrix production, the immune suppression and opportunistic infections. TGF-β1 is the main form of TGF-βand it is the most active.Smad4 is a central component of the TGF-βsuperfamily signaling pathway. The mutations and deletions of Smad4 are seen in about 50% of pancreatic cancers.TGF-β1 and Smad4 play important roles in human tumorigenesis. This experiment aims to detect the levels of TGF-β1 and Smad4 in serum and Smad4 expressive profiles in patients with ovarian tumors and normal ovaries in order to provide some experimental reference for the pathogenesis, diagnosis, monitoring, treatment, and prognosis of ovarian cancer.Materials and Methods: Enzyme-Linked Immunosorbent Assays is used to detect the serum levels of TGF-β1,Smad4 and Immunohistochemistry is used to detect Smad4 expressive profiles in patients/women with ovarian cancer, borderline ovarian tumor, benign epithelial ovarian tumor ,ovarian sex cord-stromal tumor and the normal ovaries.Results:1、The preoperative serum levels of TGF-β1 in patients with ovarian cancer show no statistical difference with borderline ovarian tumor and ovarian sex cord- stromal tumor(P>0.05),but they are all significantly higher than the serum levels of TGF-β1 in patients with benign epithelial ovarian tumor and the normal ovaries, which are statistically significant difference(P<0.01). The preoperative serum levels of TGF-β1 in patients with benign epithelial ovarian tumor are significantly higher than the normal ovaries, which are statistically significant difference (P<0.01). After operations, the serum levels of TGF-β1 in patients with ovarian cancer, borderline ovarian tumor, benign epithelial ovarian tumor ,ovarian sex cord-stromal tumor and the normal ovaries are all significantly higher than the preoperative serum levels, which are statistical difference (P<0.05) ,but they all show no difference(P>0.05). The preoperative serum levels of TGF-β1 in patients with ovarian cancer are related to histological type and the volume of ascites , which are statistical difference (P<0.05).2、The preoperative serum levels of Smad4 in patients with ovarian cancer, borderline ovarian tumor, benign epithelial ovarian tumor and ovarian sex cord- stromal tumor are higher than the serum levels of Smad4 in patients with the normal ovaries , which are statistical difference (P<0.05). After operation , the serum levels of Smad4 in patients with ovarian cancer are lower than the serum levels of Smad4 in patients with benign epithelial ovarian tumor and ovarian sex cord- stromal tumor, ,which are statistical difference (P<0.05). The postoperative serum levels of TGF-β1 in patients with benign epithelial ovarian tumor and the normal ovaries are all higher than the preoperative serum levels, which are statistical difference (P<0.05). The preoperative serum levels of Smad4 in patients with ovarian cancer are related to histological grade, histological type and the volume of ascites (P<0.05).3、The expression of Smad4 protein in ovarian epithelial tissues is lost in 38 ovarian cancer tissues(38/82,46.3%) , in 8 borderline ovarian tumor tissues (8/20, 40.0 %),in 14 benign epithelial ovarian tumor tissues(14/24,58.3%),in 4 ovarian sex cord– stromal tumor tissues(4/16,25.0%), and in 2 normal ovaries tissues(2/16,12.5%). Of 82 ovarian cancer specimens,14 are ovarian mucous adenocarcinomas and the expression of Smad4 protein is lost in 12 ovarian epithelial tissues (85.7%); Of 20 borderline ovarian tumor specimens,12 are mucinous ovarian tumors and the expression of Smad4 protein is lost in 6 ovarian epithelial tissues (50.0%); Of 24 benign epithelial ovarian tumor specimens,14 are mucinous cystadenoma of ovary and the expression of Smad4 protein is lost in 10 ovarian epithelial tissues (71.4%).There are benign, borderline and malignant lesions simultaneously in two cases of mucinous ovarian tumors and Smad4 protein is of deficiency expression in above-mentioned lesions. The expression of Smad4 protein is absent in the epithelial tissue of ovarian cancer is higher than the normal ovaries ,which is statistical difference (P<0.05).The expression of Smad4 protein is absent in the epithelial tissue of benign epithelial ovarian tumor is higher than ovarian sex cord-stromal tumor and the normal ovaries ,which is statistical difference (P<0.05).In 82 cases of ovarian cancer, the expression of Smad4 protein is lost in 20 ovarian interstitial tissues (24.4%), but the expression of Smad4 protein in ovarian interstitial tissues is lost in no borderline ovarian tumor tissues, benign epithelial ovarian tumor tissues, ovarian sex cord-stromal tumor tissues and the normal ovaries tissues. In ovarian cancer ,loss of Smad4 protein expression in the epithelial tissues is related to age, histological type, clinical stage, the volume of ascites and pelvic lymph node metastasis (P<0.05). In ovarian cancer ,loss of Smad4 protein expression in the interstitial tissue is not related to all of the clinicopathological factors that we mentioned in this paper(P>0.05).Conclusions: 1、The elevated serum levels of TGF-β1 and Smad4 in patients with ovarian cancer and ovarian sex cord-stromal tumor can be used as the biological indicators for the diagnosis. 2、The elevated serum levels of TGF-β1 and Smad4 in patients with ovarian cancer can be used as the poor prognostic indicator.3、The serum levels of Smad4 may come from the ovarian epithelial tissue and interstitial tissue in patients with ovarian cancer and ovarian sex cord- stromal tumor.4、The loss of Smad4 protein expression in ovarian cancer may be one of the important reasons for the obstacle of TGF-β1-Smad4 signal transduction and may be considered as one of the poor prognostic indicators for some patients.5、The expression of Smad4 protein can not be used as the biological indicator for the differential diagnosis of primary and metastatic ovarian mucous adenocarcinomas.6、The loss of Smad4 protein expression in ovarian cancer indicates that Smad4 may has potentially biological therapy value for some patients.更多还原