The Expression And Anti-apoptosis Of Epo In Ovarian Carcinoma

Ovarian carcinoma is the leading cause of death from gynecological cancer. Although advances in treatment with cytoreductive surgery and chemotherapy have improved the survial rate in the last decade, in most patients, the disease can not be diagnosed at an advanced stage, and survial is still poor, with a 5-yr survial rate of only 25%~30%.Erythropoietin (EPO), which is produced by the liver in the fetus and by the kidney in the adult, is the primary stimulator of red blood cell formation. Up to now, however, EPO has been accepted as an multiple functional cytokine. Over the past decade an increasing number of studies have supported the use of recombinant human erythropoietin (epoetin) in cancer patients, suggesting that it improves haemoglobin concentrations for some, which may lead to improvement in quality of life for cancer patients. In contrast, some researchers has found that EPO mimetic peptide can promotes angiogenesis and tumour cell survival, in 2003,which was published in the jounal of Lancet and Carcinogenesis.In order to determine whether EPO can be used in the patients with ovarian carcinoma ,whose haemoglobin concentrations are lower , we studied the expression and biologic effect of EPO in the main kinds ofovarian cancer. AIM:1. Detect the expression of EPO in the main kinds of ovarian carcinoma and the normal ovary tissue.2. Study the relationship between hypoxia and the expression of EPO in 3AO cell line.3. Study the biologic effect of EPO by administrating rHuEPO into 3AO cell line.METHODS:1. Detect the protein expression of EPO in Human ovrian carcinoma Tissue Array by immunohistochemistry.2. Detect the protein and mRNA expression of EPO in 3AO cell line,which was put into anoxic enviroment, by in situ hybridization and immunohistochemistry.3. Draw the growth curve of 3AO cell line in anoxic enviroment by MTT.4. Detect the apoptosis rate of 3AO cell line in anoxic enviroment by FCM.5. Detect the expressions of caspase-3 and bcl-2 in 3AO cell line, after the administration of rHuEPO ,by fluorescent immunohistochemistry.6. Detect the apoptosis rate of 3AO cell line in anoxic enviroment, after the administration of rHuEPO ,by FCM .7. Study the Ultrastructure of 3AO cell after the administration of rHuEPO.RESULTS:1. 2(of 10) control samples ,7(of 18) benign tumor samples, and 8 (of 13) borderline tumor samples were positive in EPOprotein ,but 97(of 107) ovarian carcinoma were positive(94.2%).The positive rate of III~IV stage by FIGO standard of ovarian carcinoma was 79%, which was higher than I ~ II stage significantly.2. The expression of EPO peptide in 3AO cell line cultured in anoxic environment was higher than those cultured in normal environment(P<0.05).3. At 6h point cultured in anoxic environment, the growth ability of 3AO cell decreased, but recovered nearly at 16h point. Therefore ,we suggested that ovarian carcinoma could tolerate temporary hypoxia.4. The apoptosis rate of 3 AO cell line, which was cultured anoxic environment and adiministrated soluble EPO receptor, was higher than the control group at 6~16h, significantly (P <0.05).5. Carboplatin could induce the apoptosis in 3AO cell, which displayed the lower expression of caspase-3 but higher expression of bcl-2 than the control group.6. After administration of EPO peptide, the apoptosis rate, induced by carboplatin , decreased more markedly than the control group.( P<0.05)7. The ultrastructure of 3AO cell, which was administrated carboplatin, displayed typical apoptosis morphological changes.CONCLUSIONS:1. The expression of EPO peptide in ovarian carcinoma was higher than normal ovary tissue and benign ovary tumor, and positively correlated with the FIGO stage of carcinoma(r=0.498, P<0.01).