Study On Serum CA-125 Levels And Related Genes Of Deep Infiltrating Endometriosis

Backgrounds:The pathogenetic process of endometriosis(EMs)is still unclear,which may be influenced by genetic factors.At present,there is no accurate non-invasive method for early diagnosis of EMs.Serum cancer Antigen 125(CA-125)is a biomarker that has been widely used in clinical practice to indicate EMs,but studies on its influence factors are still insufficient.Although increasing arguments on whether various EMs subtypes differ in mechanisms of pathogenesis have shown,many previous studies have not specified EMs to each subtype,including superficial EMs,ovarian endometrioma(OMA)and deep infiltrating EMs(DIE),to filter subtyped-specific candidate genes.Objectives:To analyze serum CA-125 levels of DIE cases and evaluate some influence factors of serum CA-125 levels among EMs cases,including ages at first diagnosis,rASRM(the revised American Society for Reproductive Medicine)stages,location of endometriosis lesions.To screen both subtype-specific and two-subtypes-shared related genes and pathways of DIE and OMA.Methods:The first part is a reprospective study,including data from 63 DIE cases and 63 OMA cases,involving time of first diagnosis as EMs,complication of uterine fibroids or adenomyosis,reproductive history,serum CA-125 levels tested during non-menstrual period after discontinuing oral hormone therapy≥ 3 months,and location of EMs lesions through examination of laparoscope.Compare difference between characteristics of DIE group and OMA group,and build a binary logistic regression model to investigate the influence of factors enrolled on serum CA-125 levels,with elevated CA-125 defined as CA-125≥35 U/mL(Units/milliliter).On the second part,we sequence compared ectopic and eutopic samples from 8 DIE cases through Illumina nova pe150 and use DESeq2 to screen DEGs(Differential Expression Genes).Use RT-qPCR(Reverse Transcription Quantitative Real-Time-Polymerase Chain Reaction)to evaluate mRNA levels of C1S,C1QC,C1QB to validate results of sequenced DIE data.On the third part,we download data of GSE7305,GSE25628,GSE23339 from GEO,choose eutopic or non-EMs control samples and ectopic samples to filter genes of DIE samples and OMA samples to analyze related pathways and modules.Build PPI(Protein Protein Interaction)network and screen important genes.Upload the four DEGs lists to Metascape to do a meta-analysis of multi-gene lists.Results:1.DIE group showed a higher morbidity of both uterine myoma and uterine adenomyosis than OMA group(for uterine myoma,OMA vs DIE,0.0%vs 14.3%,P=0.003;for uterine adenomyosis,OMA vs DIE,3.2%vs 22.2%,P=0.002).2.Through a binary multi-variable logistic regression analysis,19-29yr group showed significant difference(ref=40-62yr group;OR,3.492;95%CI,1.202-10.144;Wald χ2=5.279;P=0.022),rASRM StageⅣgroup also showed significant difference(ref=rASRM Stage Ⅲ;OR,2.869;95%CI,1.284-6.410;Wald χ2=6.601;P=0.010).3.There was a tendency among DIE group,serum CA-125 levels of those patients with lesions on ligaments(71.9±58.2 U/mL)were higher than those without lesions on ligaments(50.9±39.0 U/mL).4.Complement activation-related pathways showed significant difference in DIE,and related genes includes C3,CLU,C7,C4BPA,CIS,SERPING1,C4B,PROS1,C1R,C1QC,C1QB,etc.5.The key DEGs screened by MCODE shared by the DEGs lists of four data series were C3,PNOCy MYH11,MYL9,GPC3.Related GO modules include circulatory system process(GO:0003013),leukocyte migration(GO:0050900),acute inflammatory response(GO:0002526),blood vessel development(GO:0001568),embryonic morphogenesis(GO:0048598),etc.Conclusions:1.Complication of uterine fibroids or adenomyosis might be more common in DIE subtype than in OMA subtype.2.EMs patients’ serum CA-125 levels tested during non-menstrual period might be influenced by ages and rASRM stages.3.Pathways related to complement activation show potential influence on DIE.4.C3,PNOC,MYH11,MYL9 and GPC3 might participate in the occurrence and development of both DIE and OMA.