【作者】 陈芳芳；

【导师】 柳忠辉；

【作者基本信息】 吉林大学 ， 免疫学， 2007， 硕士

【摘要】 肿瘤是严重威胁人类健康和生命的疾病之一,其发生、发展是多因素、多步骤、多阶段的复杂过程,不同分期、分型肿瘤选择的治疗方式不同,早发现、早治疗是肿瘤诊治的重要原则。卵泡抑素(follistatin,FS)是一种单链糖蛋白,具有广泛的组织分布及多种生物学活性。粘蛋白1(MUC1)是mucin粘蛋白家族成员,存在于正常腺管上皮细胞及其来源的肿瘤细胞中。FS、MUC1在肿瘤诊断、预后判断等方面具有重要意义。为进一步探讨FS及MUC1在肿瘤诊断方面的临床意义,本实验利用双位点ELISA法检测肿瘤患者外周血中FS蛋白水平变化;荧光定量RT-PCR检测不同类型肺癌组织、肺癌及乳腺癌患者外周血循环FS及MUC1 mRNA水平变化;采用免疫组织化学染色检测FS、MUC1在不同病理类型卵巢癌、乳腺癌及肺癌组织内的分布情况。研究结果显示,卵巢癌、肝癌、乳腺癌、肺癌患者外周血FS蛋白水平明显高于健康对照组;除小细胞肺癌外周血MUC1 mRNA无明显变化外,肺癌组织和外周血循环FS mRNA及外周血MUC1 mRNA水平明显高于对照组,p<0.01;乳腺癌患者外周血MUC1 mRNA水平明显高于对照组,p<0.05。免疫组化研究结果表明,卵巢肿瘤组织抗FS抗体组化染色强度依次为粘液性癌>子宫内膜样癌>鳞癌>透明细胞癌>浆液性囊腺癌,抗MUC1抗体组化染色强度各肿瘤间无明显差异;肺癌组织抗FS抗体组化染色强度依次为腺癌>腺鳞癌>鳞癌>未分化小细胞癌。为研究FS、MUC1与妊娠的关系,本实验同时观察了妊娠早期及难免流产患者外周血循环FS及MUC1 mRNA水平及胎盘绒毛组织FS、MUC1表达情况。难免流产和正常妊娠比较,其绒毛组织、外周血中FS mRNA、MUC1 mRNA水平均显著下降;妊娠早期胎盘绒毛组织FS、MUC1蛋白表达水平明显高于难免流产胎盘绒毛组织。本研究资料提示FS在不同组织来源的肿瘤细胞表达水平不同,在同一组织来源的不同病理类型的肿瘤细胞表达水平也不同。肿瘤患者外周血FS、MUC1 mRNA水平升高,可能直接来源于肿瘤细胞,对肿瘤诊断具有重要意义。绒毛组织的FS及MUC1水平变化提示FS、MUC1与生理病理妊娠有关,可能在维持正常妊娠中具有重要作用,但其机制有待进一步研究。更多还原

【Abstract】 Follistatin is a single polypeptide chain, which was isolated from ovarian follicular fluid and directly inhibit the secretion of FSH from anterior pituitary, and has a widely histological distribution. The characterization of FS expression indicates that it may have different functions on different kind of cells. FS can specifically bind activin, directly inhibit activin’s biological activity and take part in regulation of physiological functions. Mouse gene knock indicates that FS plays important role from embryo to maturity of organism. FS and activin often coexist and coexpress to make a balance system for maintaining normal function of organization and organ. That is to say one is over expression or another decrease will result in imbalance of FS-ACT system with physiological defect and the course of pathology of some organization tissues and organ.MUC1 is the mucin that is a membrane protein composed of core peptide and carbohydrate side chain expressed on normal epithelia cells and adenocarcinoma cells. There are some differences of MUC1 expression between normal and tumor cells. Normal MUC1 exists on ductal epithelia as a large heavily glycosylated protein is expressed only on the apical cell surface near the duct lumen. The expression of MUC1 is up-regulated on tumour cells where it undergoes changes in deglycosylation and distributes on the whole tumor cell surface,that make it distinct from normal mucin. Therefore,the differences of MUC1 expression between normal and tumor cells make it a potential marker for tumor diagnosis.1. ObjectiveTo investigate the expression of FS and MUC1 in Human tissues and their clinical significance for diseases diagnosis.2. Methods(1) The level of FS proteins in sera of patients with cancer was assayed by two site ELISA.(2) FS and MUC1 mRNA in tissues and the peripheral blood of different patients, such as lung tumor, breast cancer, was tested by fluorescent-quantitative RT-PCR.(3) The distribution of FS and MUC1 proteins in Lung tumor, breast and ovarian cancer tissues was analysed by immunohistochemistry.(4) FS and MUC1 mRNA in the peripheral blood and placenta of pregnant woman was examined by fluorescent-quantitative RT-PCR.(5) The distribution of FS and MUC1 proteins was examined in placenta tissues by immunohistochemistry.3. Research results(1) The levels of FS protein in sera of lung cancer, breast carcinoma, liver cancer and ovarian cancer patients were greatly higher than that of the normal health group, The positive rate is ovarian carcinoma(60%)>liver cancer(40%)and breast cancer(40%)>lung cancer(30%)>gastric cancer(20%) >renal carcinoma(0%)。(2) The levels of FS protein were distinct in different type of lung cancer tissues, adencarcinoma is significantly higher than the normal group, p<0.01. There was no change in small cell lung cancer group.(3) In lung cancer patients, the level of circulating MUC1 mRNA in sera was significantly higher than the control group (p<0.01), the level of circulating FS mRNA was significantly higher than the control group (p<0.01) exclude small cell lung cancer. The level of circulating FS mRNA had not obviously changed in the cancer of breast, but the level of MUC1 mRNA was remarkably higher than control group.(4) Immunohistochemistry staining showed that FS protein expressed differently in lung cancer, adenocarcinoma>adeno and squamo cancer> squamocarcinima>small cell lung cancer.(5) FS and MUC1 protein were expressed obviously in ovary tumor tissuses. Intensity of FS is as follow: mucinous carcinoma>serous cystadenoma>endometrioid carcinoma > squamo cancer > clear-cell carcinoma > serous cystadenocarcinoma. Intensity of MUC1 has no significant difference in ovary tumor tissue.(6) The levels of FS and MUC1 mRNA in placental villus and peripheral blood were significantly decreased in inevitable abortion patients, compared with that in the subjects of the early trimester of pregnancy. Immunohistochemistry staining showed that the expression of FS and MUC1 was more significant decrease in placental tissues in inevitable abortion pateints than that in early trimester of pregnancy. 4. ConclusionThe level of FS protein is distinct in diverse tumor,and the quantity of FS, MUC1 protein and mRNA is distinct in different types of tumors come from the same tissue. All the data suggested that circulating FS and MUC1 in circulation blood may be directly secreted from tumor cells.All the findings suggest that FS expression of tumor cells and Circulating FS mRNA levels have important diagnosis value plays an important role in clinical diagnosis and determine phathological types of different cancer. However, it still need further discussion to determine whether abnormal high levels of FS and MUC1 mRNA was the cause of tumor genesis or the result of tumor genesis.更多还原