Study On The Detection Of Exosome Protein Markers In Epithelial Ovarian Cancer

Objective:Ovarian cancer is one of the common Ovarian malignancies in the female reproductive system,the mortality rate of gynecological tumor is the first,its insidious,early without typical symptoms,clinical discovery is mostly in the middle and late stage,high mortality,poor prognosis,serious threat to women’s lives.Epithelial ovarian cancer(EOC)is the most common ovarian cancer pathological types,80-90% of ovarian cancer,and serous ovarian cancer is the most common.According to statistics,70% of patients with serous ovarian cancer are at advanced stage of treatment.The purpose of this study was to extract epithelial ovarian cancer exosomes,and to detect the levels of Ep CAM CD44 and CD9 the protein markers of exosomes in epithelial ovarian cancer by flow cytometry.To verify the feasibility and practicability of the experimental method in the study of exosome protein markers of epithelial ovarian cancer.It provides a basis for the establishment of the detection system of exosome protein markers in epithelial ovarian cancer.Furthermore,a multivariate logistic regression risk prediction model was established to detect and predict early ovarian cancer.It can improve the early detection rate of epithelial ovarian cancer,assist the clinical diagnosis of ovarian cancer,and improve the survival rate.Methods:In this study,thirty-one patients with epithelial ovarian cancer who were admitted to the obstetrics and gynecology department of sichuan mianyang central hospital from August 1,2016 to November 1,2018,and who had not received radiotherapy,chemotherapy or tumor immunotherapy,were selected as the study group.Postoperative FIGO staging was 9 cases in stage Ⅰ,16 cases in stage Ⅱ and 6 cases in stage Ⅲ A.Forty patients with benign ovarian tumors(serous or mucinous cystadenoma of ovary or teratoma of ovary or chocolate cyst of ovary,who had not taken estrogen and progesterone drugs for nearly 6 months before surgery).The control group included 40 patients who underwent physical examination(none of whom had taken estrogen for nearly 6months).4ml of intravenous blood was extracted from the patients in the all groups,and plasma was extracted.Extraction of plasma-derived exosomes from exosome extracts.Detection of exosome extraction by electron microscopy,and flow cytometry was used to detect the Ep CAM,CD44 and CD9 levels of exosome protein markers,and the differences between groups were compared.All the data were statistically analyzed by SPSS22.0 software,the measurement data were expressed by x ±sem,t-test and anova were used for comparison between groups,and significance analysis was performed by holm-sidak method,with alpha=5.000%,P<0.05 was considered statistically significant.Results:Electron microscopy showed that the exosomes extracted by exosome extraction solution had a complete membrane structure with uneven size and a diameter of about 30 ~100 nm.Statistical comparison of flow cytometry showed that plasma Ep CAM+ %,CD44+ % and CD9+ % in patients with epithelial ovarian cancer were higher than those in the healthy control group with benign ovarian disease,and the difference was significant(P<0.05).There was significant difference in plasma Ep CAM+ % CD44+ % between the benign ovarian disease group and the healthy control group(P<0.05).There was no significant difference in plasma CD9+ % level between the benign ovarian disease group and the healthy control group(P>0.05).Although the plasma Ep CAM+ % and CD44+ % in the benign ovarian disease group was significantly different from that in the healthy control group.However,the plasma levels of Ep CAM+% and CD44+% in patients with epithelial ovarian cancer were(95.22 ± 0.311)% and(92.62 ± 0.409)%,higher than the plasma levels of Ep CAM+% and CD44+% in patients with benign ovarian tumors(91.68 ± 0.375)% and(89.46 ± 0.428)%.Conclusion:This study was based on the extraction of epithelial ovarian cancer exosomes and the detection of protein markers.The expression level of Ep CAM CD44 and CD9 in epithelial ovarian cancer was higher than that in benign ovarian group and healthy control group.Although the expression level of Ep CAM CD44 was significantly different between the benign ovarian tumor group and the healthy control group,however,the expression level in the cancer group was significantly higher than that in the benign tumor group.Compared with the existing methods of exosome extraction and surface marker detection,the experimental samples obtained in this research system are simple and reusable,In this research system,the acquisition of simple and reusable experimental samples has obvious advantages.The obtained exosomes can be repeatedly detected by flow cytometry to detect different tumor markers and determine their contents.Therefore,this experimental method is applicable for the extraction and detection of exosomal proteins in epithelial ovarian cancer.In the later stage,multiple protein markers were detected through the experimental method of this study,and the protein marker system of epithelial ovarian cancer was established.A multivariate logistic regression risk prediction model was established for the detection and prediction of ovarian cancer.