The Ligand Proteomics Study Of 14-3-3 Zeta In The Pathogenesis Of Ectopic Endometrial Stromal Cells In Adenomyosis

Objective: According to our previous study,we found that expression of14-3-3?is significantly higher in endometriosis ectopic tissue than that in control tissue,but whether it can really promote the occurrence and effects of adenomyosis mechanism is unclear.We plan to confirm whether high expression of 14-3-3? can promote adenomyosis occurrence by cell function experiments;and confirm the interaction of ligand protein group by TAP purification and protein mass spectrometry,select the 14-3-3 zeta differences ligand proteome of ectopic endometrial stromal cells.Using Co-IP method to confirm the interactions of 14-3-3 zeta and its ligand protein MAP3K2.Method: Culture and identify the endometrial stromal cells extracted from adenomyosis patient;Determine the expression of corresponding mRNA and protein by Western blot and RT-PCR;Construct 14-3-3? siRNA to infect the adenomyosis endometrial stromal cells,determine the apoptosis by flow cytometry,determine the cell invasion migration by Tranwell chamber analysis and confirm the effects of14-3-3? high expression for adenomyosis.Usin TAP method to analyse 14-3-3?ligand protein of endometrial stromal cells;Confirm differently high expression of14-3-3?ligand protein by mass spectrometry;Confirm the interactions between the14-3-3?and the ligand protein MAP3K2 by Co-IP.Results: 1.WB results showed that 14-3-3?protein expression in ectopic endometrial stromal cells is higher than that in eutopic cells;2.Under the interference of siRNA,the ability of proliferation,invasion and migration of ectopic endometrial stromal cells is reduced,and apoptosis is lower,the difference is statistically significant(P<0.05);3.Differential proteomes are comducted by the TAP purification combined with mass spectrometry and protein MAP3K2 is selected after data access;4.Co-IP confirmed that there exists some relationship between 14-3-3?and MAP3K2.Conclusion: 1.High expression of 14-3-3? can contribute to the adenomyosis occurrence by increasing the cell proliferation,migration,invasion,andinhibiting apoptosis;2.The 14-3-3? protein in the pathogenesis of endometrial stromal cells in adenomyosis can interact with MAP3K2,and may be associated with the occurrence of the disease.