Expression And Significances Of Oxytocin Receptors?PKC? And CPI-17 In The Uterine Junctional Zone In Women With Adenomyosis

Adenomyosis(AM)is a common gynecological diseases for childbearing age women.The disease develops in multipara 30 to 50 years old,about 50% with uterine fibroids,within about 15% with endometriosis.it is a myometrial lesion characterized by the presence of ectopic endometrial glands and stroma located deep within the surrounding myometrium(invagination of endometrium in the myometrium at a depth of at least 2.5 mm below the basal layer of the endometrium)with adjacent myometrial hyperplasia and hypertrophy.It can be either focal or diffuse(numerous foci spread throughout the myometrium)and it is often asymmetric.Nearly two-thirds of patients with clinical manifestations,Symptoms are commonly reported as menorrhagia,dysmenorrhea,the uterus increased gradually,pelvic pain,infertility and bulk-related symptoms.which has a severe impact on life quality and psychological health.The pathogenesis of AM remains unclear,Although it may be related to hormone disorder,disorder of immune factors and the imbalance of genetic material.With MRI examination in the application of disease of department of gynaecology,the pathogenesis of AM recently focused on the uterine junctional zone(JZ),it was introduced in this manner to describe this interface observed on MRI: a distinct low signal on T2-weighted sequences separating the endometrium in high signal intensity from the outer myometrium in intermediate signal.In the normal period of women of childbearing age,a JZ thickness with MRI is about 2 to 8 mm.Generally,a JZ thickness of greater than 12 mm is the most widely accepted criterion in establishing the presence of AM.The rhythmic contraction originating from JZ maintain the normal structure and function of the uterine.Current studies suggest that AM may be related to the abnormal contraction of JZ.But the specific mechanism is still unkonown.The oxytocin receptor(OTR)is a member of the class I family of G protein-coupled receptors,regulating uterine smooth muscle contraction.Protein kinase C(PKC beta)participate in the regulation of cell growth,proliferation,differentiation,apoptosis and so on a variety of signaling pathways.Protein kinase C-potentiated phosphatase inhibitor of 17 kDa(CPI-17)is main effect the substrate of PKC?,which is also the important molecular regulating calcium sensitivity.This study attempts to explore the expression of OTR,PKC??CPI-17 in the JZ in women with AM and its relationship with the onset of AM.ObjectiveUsing the immunohistochemical method and real-time fluorescent quantitative PCR detection respectively OTR,PKC beta,CPI-17 protein and mRNA expression level in uterine junctional zone of the two groups and its correlation,in order to provide new reference basis for innovative treatment research of AM.Materials and Methods1.From May.2015 to Dec.2015.45 patients with adenomyosis as study group(from 34 to 50 years old,an average of 45.6 years old)matched with 45 women with cervical intraepithelial neoplasia(CIN)III Treated by hysterectomy as control group(from 35 to 55 years old,an average of 42.2 years old)were enrolled in this study in the Third Affiliated Hospital of Zhengzhou University.There was no statistically significant difference ages.All the samples based ethics committee approval and patients' informed consent,and the pathological results should be strictly confirmed by pathology doctors.2.Methods:Immunohistochemistry and fluorescence quantitative PCR methods were used to detect the expressions of OTR,PKC beta,CPI-17 protein and mRNA in uterine junctional zone of the two groups and its correlation,we operated strictly according to the kit instructions.3.Statistics:Using SPSS21.0 statistical software to analysis datas,quantitative datas were presented in the form of x�s,different groups were compared by independent-sample t test and Wilcoxon rank sum test,Immunohistochemical qualitative datas were analyzed by Chi-Square Tests and Fisher's exact probabilities,Pearson correlation method was used to analyze the quantitative variable date correlations.The association of classified variable data was analyzed by Pearson chi-square independence test,Inspection standard ?=0.05.Results1.The Immunohistochemical results :The positive expression rates of OTR protein in JZ of AM group and control group were 77.8% and 17.8%,the difference had statistical significance(P<0.05);The positive expression rates of PKC? protein in JZ of AM group and control group were 84.4% and 22.2%,the difference had statistical significance(P<0.05);The positive expression rates of CPI-17 protein in JZ of AM group and control group were 71.1% and 60.0%,which didn't show statistical difference(P>0.05);The positive expression rates of phosphorylated CPI-17 protein in JZ of AM group and control group were 82.2% and 20.0%,which reached statistical difference(P<0.05).2.The results of fluorescence quantitative PCR showed that the relative expressions of OTR mRNA in JZ of AM group and control group were 1.437�0.605 and 0.879�0.293,which reached statistical difference(P<0.05);The expression of PKC?mRNA in JZ of AM group and control group were 1.775�0.874 and 0.943�0.289,which reached statistical difference(P<0.05);The expression of CPI-17 mRNA in JZ of AM group and control group were 1.166�0.876 and 0.994�0.382,which didn't show statistical difference(P>0.05).3.The correlation between OTR ?PKC?and phosphorylated CPI-17 in JZ of AM groupThe Immunohistochemical results showed that the correlation between OTR and PKC?was positive(r=0.453,P<0.05)in JZ of AM group,The expression of PKC? protein had positive correlation with phosphorylated CPI-17 protein expression in JZ of AM group(r=0.516,P<0.05).The fluorescence quantitative PCR results showed that the correlation between PKC?and phosphorylated CPI-17 was positive in JZ of AM group(r=0.617,P<0.05).Conclusion1.OTR?PKC?and phosphorylated CPI-17 were highly expressed in in JZ of AM group,suggesting that it might be involved in the development of adenomyosis.2.OTR and PKC? were positively correlated,PKC? and phosphorylated CPI-17 were positively correlated in JZ of AM,indicating that OTR activate PKC?/CPI-17 signaling pathway to enhance Ca2+ sensitization,it might be involved in the development of adenomyosis.