The Detection Of Peripheral Blood Circulating FS And MUC1 MRNA In Human And Its Application

FS (Follistatin, FS) is a single polypeptide chain glycoprotein isolated from ovarian follicular fluid which directly inhibits the secretion of FSH from anterior pituitary cells. FS has a widely histological distribution especially has a high expression in ovary, pituitary, kidney and embyro which indicates that FS has a different function on different kind of cells. As an activin (ACT) binding protein, FS participates in various kinds of physiological and pathological functional regulation by blocking the biological activities of ACT. The studies on gene–knocked out mice suggest that FS is involved in the process from embryogenesis to maturity. FS and ACT co-exist and express in one organ as a balance system to maintain its normal function. The high expression or low expression of either of FS and ACT can lead to functional unbalance of the organ. Current studies suggest that FS plays an important role in diseases including hepatic fibrosis, pulmonary fibrosis, renal injury, skin injury, brain injury, atherosclerosis and so on. Mucins-1 (MUC1) is a kind of macromolecule glucoprotein which widely distribute on the surface of mucous membrane in several kinds of tissues and organs. 13 different kinds of MUC have been found (MUC1-MUC13). MUC1 mucoprotein normally expresses in epithelial cell proximal end and glandular cavosurface which is polarly distributes in various kinds of tissues, organs. Abnormity expression of MUC1 is involved in many kind of cancer so that MUC1 are regarded as a cancer marker. In recent years, numerous vaccines based on MUC1 immunogen and immune targeting vector based on MUV1 antigen has been used to cancer biotherapy research and many have been used in clinical experiment phase.1. ObjectiveTo investigate the application value of FS, MUC1 mRNA examination in clinical disease diagnosis2. Methods(1) Flurescent quantity RT-PCR was used to examine the peripheral blood circulating FS and MUC1 mRNA of several diseases, such as cancers (lung cancer, ovarian carcinoma, liver cancer, breast cancer, renal carcinoma and benign tumor), non-tumors (diabetes mellitus, hypertension, acute myocardial infarction, pneumonia, pulmonary tuberculosis) and normal pregnant women and threatened abortion patients(2) Immunohistochemistry stain was used to examine the FS and MUC1 protein expression on ovarian carcinoma, cancer of breast, lung cancer and placental villus tissue.(3) Two site ELISA was used to examine the expression of mature protein expressions of FS and MUC1 in healthy adults and pregnant women peripheral blood.3. Results(1) The results showed that the level of circulating FS mRNA in sera of the diabetic among the benign disease groups had been detected obviously increased. There's a statistical significance (p<0.01) compared with that of the healthy control group, while other benign disease groups possessed no statistical difference with the healthy adults group (p>0.05). The peripheral blood circulating FS mRNA level in the ovarian carcinoma and lung cancer patients in the cancer group had been detected obviously increased and there's a statistical significance compared with that of healthy control group (P<0.01). The peripheral blood circulating FS mRNA level in healthy old-aged group increased slightly, whereas there is no statistical difference compared with the healthy youth group (p>0.05). The peripheral blood circulating FS mRNA level in early and late pregnant group women had no obvious difference (p>0.05). The peripheral blood circulating FS mRNA level in threatened abortion patients increased evidently. There is a statistical significance (p<0.05) compared with normal early pregnant group. The peripheral blood circulating FS mRNA level examined by ELISA in late pregnant women are obviously higher than that in early pregnant group (P<0.01). The above data revealed that the peripheral blood circulating FS mRNA level was not closely related with age and pregnancy, but played much more significant role in cancer diagnosis than the examination of FS protein in peripheral blood. (2) Investigation of the peripheral blood circulating MUC1 mRNA in various diseases patients demonstrated that the MUC1 mRNA level of myocardial infarct patients in benign group increased obviously and there was a statistical significance (p<0.05) compared with the control group whereas there was no significant difference among the diabetics, hypertension, pneumonia and pulmonary tuberculosis groups. The peripheral blood circulating MUC1 mRNA level in ovarian carcinoma, lung cancer and breast cancer were obviously higher than that in control group and there were statistical significances (p<0.01, p<0.01 and p<0.05 respectively) whereas the peripheral blood circulating MUC1 mRNA level in liver cancer, stomach carcinoma, renal carcinoma and benign cancer patients had no significance changes. There was no statistic difference (p>0.05) between the peripheral blood circulating MUC1 mRNA level in healthy gerontism group and healthy adult group. Compared with early pregnant women, the peripheral blood circulating MUC1 mRNA level in late pregnant group increased obviously (p<0.01).4. Conclusion(1) The peripheral blood circulating FS mRNA level in lung cancer and ovarian cancer patients are highly expression. There is a significant difference compared with normal control group which suggests that the examination of the peripheral blood circulating FS mRNA may act as an assistant diagnosis for clinical cancer. The peripheral blood circulating FS mRNA level in adult is not closely related with age and pregnancy so that FS mRNA expression level plays a much more significant role in cancer diagnosis than the examination of peripheral blood circulating FS protein. But whether the abnormal high expression is the result of cancer genesis or cancer formation requires further investigation. The peripheral blood circulating FS mRNA expression level of threatened abortion patients increased significantly compared with normal early pregnant group which suggests that the expression of peripheral blood circulating FS mRNA may be associate with pathological pregnancy. What's more, the expression of peripheral blood circulating FS mRNA also might play certain part in diabetic diagnosis and assessment of therapy effect, but the relationship with the formation of diabetic still need further study.(2) The peripheral blood circulating MUC1 mRNA levels in ovarian carcinoma, breast cancer and lung cancer patients are obviously increased highly and there is a statistical significance which reveals that the examination of peripheral blood circulating MUC1 mRNA may give an assistant diagnosis for clinical cancer. Compared with early pregnant women, the peripheral blood circulating MUC1 mRNA levels in late pregnant group increased obviously and peripheral blood circulating MUC1 mRNA level in threatened abortion patients also increased evidently which indicates that MUC1 might have some relationship with physiological and pathological pregnancy. The research approves that peripheral blood circulating MUC1 mRNA level in myocardial infarct patients increased obviously. However, the mechanism is still unclear. Whether MUC1 could act as a kind of available marker in acute myocardial serum still or not need further investigation. (3) The experiment results show that peripheral blood circulating MUC1 mRNA level increased obviously in cancer, physiological and pathological pregnancy and myocardial infarct patients which indicates that as an assistant index for cancer diagnosis, the examination of peripheral blood circulating FS mRNA may has much more speciality than that of MUC1.