Expression And Significance Of T Cell Immunoglobulin Mucin Molecule 3 And Its Ligand Galectin-9 In Patients With Adenomyosis

Objective:Adenomyosis(AM)is one of the common gynecological diseases whose main clinical symptoms are increased menstrual flow,prolonged menstrual period,and progressive dysmenorrhea.It has seriously affected the physical and mental health of women in reproductive age.The clinical manifestations of the disease are diverse.Although histologically benign,it has malignant biological behaviors such as hyperplasia,invasion,and relapse.The pathogenesis of adenomyosis has not yet been fully elucidated.The theory of basal endometrial invasion has been accepted by scholars.In addition,related factors such as genetics,immunity,estrogen and progesterone,and viral infection also play an essential role.At present,more and more studies indicate that cellular immunity is involved in the occurrence and development of adenomyosis.T cell immunoglobulin mucin molecule 3(TIM-3),as a member of the T cell immunoglobulin and mucin domain family,is expressed on T cells,DC cells,and NK cells which an important negative immunoregulatory molecule on the cell surface.TIM-3 has an immunoregulatory effect on T cell-mediated cellular immune response,its excessive activation and important effects on different types of immune responses such as intracellular infection,tumor immune microenvironment,and autoimmune disease.Galectin-9(Gal-9)is the most important ligand of TIM-3.It is widely present on the surface of non-immune cells in the human body and also expressed on peripheral immune cells.Gal-9 expression was significantly increased under cytokine stimulation.The excessive activation of TIM-3/Galectin-9 will inhibit the immune activity of T cells in the immune microenvironment,which will lead to immune tolerance and immune escape.Besides related immune cells,more and more studies have found that this pathway was abnormally expressed in tissues of benign and malignant disease,such as melanoma,osteosarcoma,clear cell renal cell carcinoma,and cervical cancer.Methods:Eutopic endometrium and ectopic lesions from 15 patients with adenomyosis and endometrium from 13 women without adenomyosis were employed in the study.Immunohistochemistry and western blot analysis were used to detect TIM-3 protein and Gal-9 protein expression and semi-quantitative analysis.RT-PCR was used to detect the related mRNA expression levels.Results:(1)In immunohistochemical analysis,the expressions of TIM-3 protein and Gal-9 protein in eutopic and ectopic endometrial glands of adenomyosis were higher than those of the control group.There was almost negative expression of TIM-3 protein in the stroma,and Gal-9 protein levels in the eutopic and ectopic endometrial stroma of adenomyosis are not significantly different from those in the normal control endometrium.(2)In western blot analysis,the expression of TIM-3 in ectopic endometrium of adenomyosis is higher than that of normal endometrium.The expression of Gal-9 protein between the eutopic endometrium of endometriosis and the endometrium of the control group slightly missed the margin of significance.(3)In the RT-PCR method,the expression of TIM-3 and Gal-9 related mRNA in eutopic endometrium of adenomyosis is higher than that in the control endometrium.Conclusion:(1)The expression of TIM-3 protein and its ligand Gal-9 protein in eutopic and ectopic endometrium of adenomyosis is elevated,suggesting that TIM-3/Gal-9 signaling pathway may be involved in the development and progression of adenomyosis.(2)Providing a new orientation for immunotherapy of adenomyosis.