The Study On Relationship Between The Chemokine Ligand 18 Gene And Ovarian Cancer

Malignant ovarian tumors has its high mortality rate,complex biological characteristics,and low 5-year survival rate due to its low rate of early detection, easy to replase,and the difficulty of treatment in advanced stage,which led ovarian cancers to the most threatern of female genitai malignant diseases.At present,CA125 is widely used in diagnosis of carcinomar with its deficiency in clinical practice which shows that apply CA125 a single indicator has low specificity and sensitivity,that means unsatisfactory on diagnosis.But other serum markers studied still can not replace CA125.Besides,imaging examinations and surgerys can't used for early dianosis because of high cost and serious trauma.Therefore,it is urgent to find a more credible tumor marker to improve the rate of early diagnosis.Chemokine C-C motif ligand 18 is an inflammatory small secretion molecular chemotaxis protein,but recent studies have shown that,CCL18 may play a partial anti-tumor role of immune-regulating,thus affecting the tumor invasion,metastasis and other biological behavior and prognosis of patients.We have analysised serum different protein express profile in malignant ovarian tumors,benign ovarian tumors and normal ovarian by SELDI-TOF-MS combined with IMAC-Cu and WCX-2 protein microarray early,screening out of 31 potential serum marker proteins.CCL18 was one of the protein indentified and found with important diagnosis affect in malignant ovarian tumors[1].This study demonstrates chemokine ligand 18's expresstion in malignant ovarian tissues and its influence in malignant ocarian tumors' occurrence and development.In this study,the influence of CCL18 in the occurrence and development of ovarian cancer through molecular biology techniques and in vitro experiments was investigated,and the acdemic progress was obtained of follows:1,CCL18 mRNAwas detected in 31 patients with ovarian cancer,14 patients with benign ovarian tumors and 14 cases of normal ovarian tissue by real-time fluorescence quantitative PCR,the results demonstrated that CCL18 was significantly higher expressed in ovarian carcinoma(p＜0.05) than in benign ovarian tumor and normal ovarian tissue.But CCL18 expression was unrelated with clinical pathyology and survival of ovarian cancer.2,PET SUMO-CCL18 prokaryotic expression plasmid was contructed and the CCL18 recombinant mature peptide was accessed after expression and purification with genetic engineering methods.LCM combined immunomagnetic beads and MALDI-TOF was used to investigate the expression of CCL18 protein in malignant ovarian tumors using the recombinant peptide for standard protein.The result showed that CCL18 protein was expressed in malignant ovarian tumors,and the proportion of CCL18 in ovarian cancer was high(9/10),benign ovarian epithelial cells had only rates of 50%, normal ovarian epithelial cells had no significant expression of CCL18 protein.3,CCL18 was detected in epithelial ovarian cancer cell lines cultured in vitro.CCL18 eukaryotic expression vector was built to study the function of CCL18 in ovarian cancer.Cell cycle in SKOV3-CCL18 proliferation group(S+ G2+M) was 32.8%,which was an obvious upward trend compared with the control group(p＜0.05);The growth curve had no significant difference between CCL18 and control group;post-invasion transfected SKOV3 cells has significantly increased adhesion and migration(p＜0.05).4,CCL18 shRNA eukaryotic expression vector was built and infected into SKOV3 cells to study the infulence in invasion and metastasis of ovarian cancer cells in vitro.The growth curve showed there was no significant difference between CCL18 and control group;cell proliferation cycle in SKOV3-CCL18 group(S+G2+M) was 17.91%,the difference has statistical significance (p＜0.05);invasion,migration,Adhesion experiments were found to have significantly decreased.In shot,epithelial ovarian cancer cells of both cultured in vitro and captured in tissue were detected the expression of CCL18,and the quantity of CCL18 expression showed that malignant ovarian tumors was significantly higher than that in ovarian benign tumor and normal ovary.Cytology experiment in vitro showed that CCL18 had significant correlation with invasion and metastasis of epithelial ovarian cancer,which ment that CCL18 has the potential to be the marker of epithelial ovarian cancer for early diagnosis and the molecular target for therapy.