The Reverse Capture Antibody Microarray Profiling To Screen Autoantibody In Endometriosis And Epithelial Ovarian Cancer

Objective: To study if there are specific antibody biomarkers in the plasma samples of endometriosis and epithelial ovarian cancer patients.Methods: we employed an innovative'reverse capture antibody microarray'platform to profile five EMs and five OC plasma samples respectively. We used native host antigens derived from lesion tissues as'baits'for the capture of differentially labelled patient and control autoantibodies. Immunohistochemistry to determine the expression levels of target antigens.Result: Forty to fifty-seven autoantibodies that were identifid in the EMs plasma samples compared with controls. The top 20 of antibodies are orderly: APC, DLP1, PRK2, Integrinβ1, Synuclein-1, Psme3, FAS, DNA Topoisomerase I, LSP-1, TAF-172, TopoisomeraseⅡb, CA150, COL7A1, p57, JNK1, GCIP, DAP3, L-Caldesmon, TRAX and GSPT2. Eleven antibodies present simultaneously in five EMs sample. Of these, nine antibodies belong to the top 20. They are DLP1(the second), PRK2 (the third), Integrinβ1 (the forth), Synuclein-1(the fifth), Fatty Acid Synthase(the seventh), CA150(the twelfth), COL7A1(the thirteenth), p57 (the fourteenth) and L-Caldesmon(the eighteenth). Of these, six target antigen were analyzed by immunohistochemistry. Fatty Acid Synthase,PRK2,DLP1,Integrinβ1 and Synuclein-1 proteins were located in cytoplasm, while CA150 proteins were located in cellular nucleus. The protein expressions of Fatty Acid Synthase in EMs and control were 85% and 20% respectively( P<0.05) . The protein expressions of PRK2 in EMs and control were 50% and 20% respectively(P>0 .05) . The protein expressions of DLP1 in EMs and control were 45% and 10% respectively(P>0 .05) . The protein expressions of Integrinβ1 in EMs and control were 95% and 10% respectively( P<0.05) . The protein expressions of Synuclein-1 in EMs and control were 80% and 20% respectively( P<0.05) . The protein expressions of CA150 in EMs and control were 90% and 20% respectively( P<0.05) .Fifty to eighty-one autoantibodies that were identifid in the OC plasma samples compared with controls. The top 20 of antibodies are orderly: NF-ATc1, AP-180, RanBP3, IGFBP-3, NEK2, ILK, p57, DAP3, Flotillin 2, Synuclein-1, APC, Integrin b1, b-Arrestin1, HAP-1, SRP54, La Protein, p45, SCAR-1, DLP1 and COMT. Nineteen antibodies present simultaneously in five OC sample. Of these, seventeen antibodies belong to the top 20. They are NF-ATc1(the first),AP-180(the second), RanBP3(the third), IGFBP-3(the fourth), NEK2(the fifth), ILK(the sixth), p57(the seventh), DAP3(the eighth), Flotillin 2 ( the ninth), Synuclein-1(the tenth), APC(the eleventh), Integrin b1( the twelfth), b-Arrestin1(the thirteenth), SRP54(the fifteenth), p45(the seventeenth), DLP1(the nineteenth) and COMT(the twentieth). Of these, six target antigen were analyzed by immunohistochemistry. AP180, IGFBP-3, NEK2, Integrinβ1, Synuclein-1 and WAVE1 proteins were located in cytoplasm. The protein expressions of AP180 in OC , borderline caner and benign control were 85%, 80% and 70% respectively(P>0 .05 ). The protein expressions of IGFBP-3 in OC , borderline caner and benign control were 100%, 70% and 30% respectively(P<0 .05) . The protein expressions of NEK2 in OC , borderline caner and benign control were 95%, 60% and 20% respectively(P<0 .05) . The protein expressions of Integrinβ1 in OC , borderline caner and benign control were 100%, 50% and 20% respectively(P<0 .05) . The protein expressions of Synuclein-1 in OC , borderline caner and benign control were 100%, 90% and 70% respectively(P>0 .05) . The protein expressions of WAVE1 in OC , borderline caner and benign control were 100%, 80% and 30% respectively(P<0 .05) .Conclusion: The'reverse capture'antibody microarray profiling to screen autoantibody in plasma is reliable. Autoantibody profiling may have an unexpected utility in uncovering proteins which express abnormally in lesion.