【作者】 王永香；

【导师】 柳忠辉；

【作者基本信息】 吉林大学 ， 免疫学， 2006， 硕士

【摘要】 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。为了进一步探讨其在肺肿瘤方面的临床意义,采用荧光定量RT-PCR检测了肿瘤患者外周血循环卵泡抑素mRNA水平;免疫组织化学染色检测不同类型肺肿瘤细胞卵泡抑素的表达情况;利用卵泡抑素双位点ELISA进一步检测肺肿瘤细胞分泌FS影响因素。研究结果表明,肺癌及卵巢癌患者外周血循环FS mRNA水平明显高于对照组,统计学比较差异显著,分别为P<0.05,P<0.01;肝癌、胃癌、乳腺癌、肾癌及非肿瘤疾病糖尿病和甲状腺功能亢进患者血循环FS mRNA水平无明显变化。各种类型肺癌患者外周血循环FS mRNA水平不同,以未分化小细胞癌患者最高;免疫组织化学染色显示抗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;手术获得的肺肿瘤细胞体外培养可以直接分泌卵泡抑素,具有血清依赖特点;TPA能促进肿瘤细胞分泌卵泡抑素,而Forskolin无明显促进作用,显示肿瘤细胞分泌卵泡抑素与PKC信号传导途径有关。本研究资料提示肿瘤患者外周血卵泡抑素水平升高,可能来源于肿瘤细胞的直接分泌;卵泡抑素在不同类型肺癌细胞表达水平不同;肺癌患者循环FS mRNA表达增加。因此,肿瘤细胞卵泡抑素表达水平及循环卵泡抑素mRNA水平对肺癌的临床诊断、分型有重要的辅助诊断价值。更多还原

【Abstract】 Follistatin is a single polypeptide chain,which was isolated fromovarian follicular fluid and directly inhibit the secretion of FSH fromanterior pituitary cells. Follistatin has a widely histologicaldistribution, follistatin mRNA with highest expression in adult ovary、pituitary、kidney、in fetal heart and liver. The character of FSexpression indicates that it may have a different function on differentkind of cells. FS can specifically binds activin, directly inhibitactivin`s biological activity and take part in many adjusting ofphysical function. Mouse gene knock indicate: FS plays importmentrole from former of embory to maturity of organism, FS often coexistand express with activin to make a balance system for maintainingnormal growth of organization and organ. One is over expression oranother decrease will result in imbalance of FS-ACT system, and withphysiological defect and the course of pathology of some organizationand organ. In the present study, FS was involved in forming of hepaticfibrosis, lung fibrosis, renal injure, injure of skin, injure of brain,atherosclerosis, PCOS, disease of reproduction amd gonal.ACTH can be examined in the extractive of organization of lungcancer (small cell lung cancer, squamocarcinima, adeno and squamocancer, adenocarcinoma). Cytoplasm of small cell lung cancercontains neuron secretive granulae, can secrete ACTH、5-HT、kinin、histamine, at the same time, 5-HT, kinin and histamine can adjust andraise ACTH level in the serum.Cultivated cell lines of small cell lungcancer can secrete ACTH. Activin A inhibin ACTH secretion fromsmall cell lung cancer, if FS was joined into the fluid, the action willdisappered completely. All these indicated that FS-ACT havecorrelation with lung cancer, especially small cell lung cancer.1. Research objectiveTo investigate FS`s expression and application in lung cancer,possible origin of serum FS about patient with cancer, secretivepathway, influential factor and signal trsnsduction mechanism.2. Research methods(1) Circular FSmRNA of different patients such as-lung tumor(small cell lung cancer, squamocarcinima, adeno and squamo cancer,adenocarcinoma), ovarian cancer, hepatic carcinoma, gastriccarcinoma, cancer of breast, renal carcinoma and non-tumor diseasefor example diabetes、 hyperthyroidism and the normal health groupwas tested by using fluorescent-quantitative RT-PCR.(2) Lung tumor (small cell lung cancer, squamocarcinima, adenoand squamo cancer, adenocarcinoma and lung benign disease wereanazyled by immunohistochemistry.(3) To observe the origin of FS in tumor patients, we selectedorganization of tumor that obtained from operative patients with highFS, separated tumor cell and cultivate it, FS level was examined incultivated tumor cell by ELISA.(4) Using activator TPA and Forskolin that general trsnsductionpathway of signal to stimulate cultivated tumor cell.3. Research results(1) Circular FS mRNA of lung cancer and ovarian cancer is greatlyhigher than that of the normal health group, Difference is significantlyby statistics. P<0.01 and P<0.05, other cancer have no obviouschange.in lung cancer group, Circular FS mRNA of small cell lungcancer is the highest,then the adeno and squamo cancer,squamocarcinima, adenocarcinoma.(2) Immunohistochemistry staining showed that positive granulaewas distributed in cytoplasm. FS was not expressed in cytoplasm ofnormal cylindrical epithelial cell, differential tumor indicated differentFS level, positive rate is in turn: adenocarcinoma>adeno and squamocancer> squamocarcinima>small cell lung cancer>lung benign disease.(3) Serum FS level of patients with tumor is probable directlysecreted by tumor cell. Secretion of FS in tumor cells has character ofserum dose-dependent, high serum can increase FS level.(4) TPA, activator of PKC, can increase secretion of FS and hasdose-dependent. Forskolin, activator of PKA, has no obviousinfluence with FS.4. Research conclusionCircular FS mRNA levels of parients with lung cancer and ovarycancer are high expression, and difference significantly compared withnormal control group. In lung cancer, FS mRNA levels of small celllung cancer is the highest. Circular FS of patients with cancer candirectly derive from secretion of tumor cell. Secretion of FS by tumorcell has related to signaling transduction pathway of PKC.Immunohistochemistry staining showed that distribution andexpression level of FS is different in the diverse tumor, circular FSmRNA levels of patients is also markedly changing in the varioustumor. These findings suggest that FS expression of tumor cells andCircular FS mRNA levels have important diagnosis volue anddetermine types of tumor in the different lung cancer. Yet, It deservesfurther to study that abnormal high levels of FS mRNA was cause oftumorgenesis or the result of tumorgenesis.更多还原