Preliminary Study On The Predictive Value And Mechanism Of GA125,HOXA10 And HOX Site LncRNA (HOXA11-AS) In Early Infertile Patients With Endometriosis

Background Endometriosis is the presence and growth of functional endometrial tissue outside the uterine cavity.It is an estrogen-dependent disease and is closely related to pelvic pain and infertility.Endometriosis is a common disease in women of reproductive age.The prevalence of endometriosis in women of reproductiv age approaches 10%.Classic studies suggested that 25%to 50%of infertile women have endometriosis and that 30%to 50%of women with endometriosis are infertile.More than 50%of the women with normal pelvic examination was found endometriosis by the examination of laparoscopy.So far,basic experiments and clinical studies on the pathogenesis of endometriosis have not confirmed the specific mechanism of infertility caused by endometriosis,which may be related to pelvic anatomy changes caused by adhesion and immune factors.Whether the mechanism of infertility is different between mild endometriosis and severe endometriosis is not clear.Because moderate and severe endometriosis may cause pelvic adhesion,resulting in tubal distortion affects tubal peristalsis,thereby affecting the operation of fertilized eggs and resulting in infertility.The problem of infertility caused by it may be early diagnosed and recognized.However,infertility caused by mild endometriosis cannot be recognized or valued as early as possible due to mild or asymptomatic symptoms,resulting in delayed diagnosis and treatment of infertility related to mild endometriosis.How to achieve early diagnosis and treatment without excessive treatment,this has become the focus of clinical research.Therefore,a highly sensitive and specific biomarker is urgently needed clinically.We found in the clinical practice of reproductive surgery that when the serum CA125 of infertility patients was between 20 and 35U/ml,it was consistent with the diagnosis of mild endometriosis under laparoscopy.Is CA125,as a marker of ovarian cancer,different from the serum detection threshold of 35U/ml in predicting endometriosis related infertility?In addition,the relationship between endometriosis and low fertility has not been determined,and many problems are confused.For example,the fallopian tube of patients with endometriosis is mostly unobstructed.Even if it is a severe pelvic adhesion,the oviduct is mostly unobstructed.Infertility can also occur in many patients without pelvic adhesions,with very mild symptoms of endometriosis,or with no symptoms and only with mild endometriosis found during laparoscopy.Changes in immune factors should occur in all patients with endometriosis,but some patients with endometriosis can naturally get pregnant.Studies have shown that implantation failure may be one of the potential causes of infertility in patients with peritoneal endometriosis.The expression of HOXA10 is necessary for embryo implantation and endometrial receptivity.Human beings have a total of 39 HOX genes,which are divided into A,B,C and D gene clusters.Among the HOXA gene clusters,HOXA10 gene has the greatest influence on embryo implantation,which is crucial to the development of embryos and uterus.It has been reported in the literature that IncRNA HOXA 11-AS at the HOX site is predicted to regulate the expression of HOXA10 mRNA.We targeted the IncRNA HOXA11-AS at HOX site from the perspective of endometrial receptivity,and explored whether HOXA 11-AS was involved in the development and progression of endometriosis related infertility or the change process of endometrial receptivity.Objective:By detecting serum CA125 level before laparoscopic surgery combined with detection in expression of HOXA10 in eutopic endometrium,through comparing the different CA125 levels in endometriosis patients and controls of the postoperative pregnancy rate,explore the early predictive value of CA125 levels in mild endometriosis-associated infertile patients.In addition,the expression of HOX site IncRNA(HOXA11-AS)in the eutopic or ectopic endometrium of endometriosis was studied to investigate the possible mechanism in endometriosis-associated infertility.Methods:A total of 196 patients who underwent laparoscopic surgery for infertility in Yuhuangding hospital in Yantai from March 2014 to August 2016 were collected.In all 196 patients,3ml of fasting venous blood was extracted in the morning and CA125 value was measured in non-menstrual period.According to the exploration during laparoscopy,97 cases were divided into endometriosis-associated infertility group and 99 cases were divided into tubal factor infertility group as control group.Meanwhile,20 patients with peritoneal endometriosis with typical lesions in laparoscopy were selected,and 15 patients with tubal factor infertility without endometriosis were selected as the control group.After hysteroscopy exclude uterine cavity lesions,we scrape two groups of patients in endometrial tissue by liquid nitrogen quick frozen storage in refrigerator spare-80 ?,and test the HOXA10 mRNA to the two groups of patients at the same time also has carried on the serum CA125,in order to understand the influence of the receptivity of endometrium in mild endometriosis patients.We also analyzed the postoperative clinical pregnancy rate of the different CA125 levels group according to the level of CA125 and compared the postoperative clinical pregnancy rate with the control group.In addition,We choose between August 2014 and May 2015 in Yantai Yuhuangding hospital patients undergoing laparoscopic evaluation of infertility,laparoscopic peritoneal endometriosis lesions 30 cases in laparoscopic surgery,15 patients with ovarian endometriosis,the control for the 15 patients with tubal factor infertility without endometriosis.The pelvic peritoneum in typical endometriosis patients were collected separately.Patients with endometriosis of the ovary were selected from the wall of the cyst.At the same time,the uterine cavity tissue samples of the same patients were collected by scraping the uterus after hysteroscopy.In addition,the uterine endometrial tissue of infertile women with tubal factors was collected,and the endometrial tissue was collected by curettage after hysteroscopy was performed to eliminate the uterine cavity lesions.Samples collected after liquid nitrogen quick frozen,stored in the refrigerator-80 ? for later use.On the first day after surgery,5ml of fasting blood was reserved for the enrolled patients.The differential expression of HOXA11-AS,HOXA9,HOXA10,HOXA11 and HOXA13 in peritoneal,ovarian endometriosis and the control in the eutopic endometrium,ectopic endometrium and peripheral blood lymphocytes was detected by real-time quantitative polymerase chain reaction(qRT-PCR).Results(1)The comparison of preoperative CA125 between the endometriosis group and the tubal factor infertility group showed that the difference was higher in the endometriosis group than in the tubal infertility group(19.46 ± 8.47 vs 12.75 ±4.53),and the difference between the two groups was statistically significant(p<0.001).The diagnostic coincidence rate of endometriosis diagnosed by laparoscopy was increased with the increase of serum CA125.The sensitivity and specificity of the diagnosis of mild endometriosis were 56.2%and 92.0%when serum CA125 was 17.49u/ml by ROC curve.(2)Subgroup analysis found that preoperative serum CA125 level of patients with peritoneal endometriosis was significantly higher than that of the tubal factor infertility(19.58 ± 5.84 vs 11.31 ± 2.55),and the difference was statistically significant(P<0.05),While HOXA10 expression of endometrium in patients with peritoneal endometriosis was lower than that of the control group(0.1067± 0.0413 vs 0.1809 ± 0.0514),and the difference was statistically significant(P<0.05).(3)The postoperative clinical pregnancy rate of patients in endometriosis infertility with serum CA125 ?18 U/ml was lower than that of those with CA125<18U/ml(41.2%vs 65.8%)and those with CA125? 18U/ml in endometriosis infertility were lower than that of those with tubal factor infertility(41.2%vs 76.9%).The difference between the two groups was statistically significant(P<0.05).(4)The expression levels of HOXA11-AS and HOXA9,HOXA10,HOXA11,HOXA13 in the ectopic endometrium with peritoneal and ovarian endometriosis were higher than those of the eutopic endometrium,and there were significant differences between two groups(P<0.05).Two types of endometriosis were analyzed together,the results were consistent with the above.(5)HOXA10 and HOXA11 of peritoneal and ovarian endometriosis were all low expressed in the eutopic endometrium,and the expression level was significantly lower than that of the tubal infertility control group(P<0.05),while the expression levels of HOXA11-AS,HOXA9 and HOXA13 were not significantly different between the two groups(P>0.05).(6)IncRNA(HOXA11-AS)and HOXA10 mRNA are expressed in eutopic endometrium,ectopic endometrium and peripheral blood lymphocytes of patients with peritoneal and ovarian endometriosis.LncRNA(HOXA11-AS)and HOXA10 mRNA expression was highest in the ectopic endometrium of the three tissues(P<0.05).Conclusion(1)Serum CA125 greater than or equal to 18(17.49)U/ml of can be used as an early clinical predictor for infertility patients with mild endometriosis,but infertility caused by other factors should be excluded.(2)Mild endometriosis may lead to decreased fertility by affecting endometrial receptivity.Therefore,according to the perspective of endometrial receptivity and postoperative pregnancy rate,patients with mild endometriosis combined with infertility are worthy of our early detection and attention.(3)LncRNA(HOXA11-AS)may play a role in the pathogenesis of endometriosis.However,the effect of IncRNA(HOXA11-AS)in endometriosis-associated infertile patients' endometrial receptivity is very limited and needs to be further studied.