Go To The Influence Of The Methylation Of Drugs On The Adenomyosis Clinical Biological Behavior And Progesterone Receptor B

Adenomyosis is a common gynecologic disorder with a poorly understood pathogenesis.Although adenomyosis differs somewhat from endometriosis in its risk factor profile,the two nonetheless share an uncanny similarity in definition,estrogen dependence,symptomatology,and many documented molecular aberrations.These similarities may imply an identical pathogenesis.Treatment of adenomyosis has been a challenge,with hysterectomy being the treatment of choice and progestogenic agents are ineffective.Progesterone plays an important role in the regulation of normal endometrium function by binding to progesterone receptor(PR),especially PR-B.In endometriosis,the critical role of PR-B and its methylation in the pathogenesis has been more or less well-established.The delineation of PR-B and its methylation in adenomyosis may be pivotal in understanding its pathogenesis and developing better therapeutics.In this study,we investigated the relationship between the aberrant methylation of PR-B gene and clinic pathological factors of adenomyosis.PartⅠThe expression and methylation of PR-B in adenomyosis.Objectives:To investigate the expression and methylation of progesterone receptor B(PR-B) gene in eutopic and ectopic endometriun and and its effect in the genesis of adenomyosis.Methods:Ectopic and homologous eutopic endometrium from 50 women with adenomyosis and endometrium from 18 age-and menstrual phase-matched women without adenomyosis were used for immunohistochemical and methylation-specific PCR(MSP) analysis of PR-B.Results:Compared with normal endometrium,PR-B immunoreactivity were significantly reduced in ectopic as well as eutopic endometrium from women with adenomyosis(p＜0.01),methylation of PR-B in ectopic endometrium were significantly higher than normal endometrium(p＜0.05),and the methylation of PR-B in ectopic as well as eutopic endometrium from women with adenomyosis were negatively associated with the immunoreactivity(p＜0.05). Conclusions:These findings suggest that,lower expression and higher methylation of PR-B may be involved in the genesis of adenomosis.PartⅡThe effect of ADC and TSA on primary adenomyotie stromal cells derived form women with adenomyosis.Objectives:To evaluate the effect of 5-aza-2'-deoxycitidine(ADC) and Trichostatin A(TSA) in the expression of PR-B and the biological behaviour of ectopic endometrial stromal cells from women with adenomyosis in vitro.Methods:The ectopic and normal endometrial stromal cells were isolated and cultured in vitro,then exposed to different concentration of ADC and TSA.The growth and proliferation of ectopic and normal endometrial stromal cells were measured by MTT assay,the cell cycle distribution was measured by flow cytometry, the methylation of PR-B was analyzed by MSP,and the expression of PR-B was ananlyzed by Realtime PCR and Western Blot.Results:ADC and TSA could inhibit the proliferation and block the cell cycle of ectopic and normal endometrial stromal cells in a dose dependent manner,and this effect was absolutely higher than that of ectopic cells.Compared with normal endometrial stromal cells,the expression of PR-B were significantly reduced in ectopic endometrial stromal cells,and the methylation of PR-B were significantly higher(p＜0.01)。The expression of PR-B in ectopic cells were significantly increased after the treatment of ADC,and a combination of ADC and TSA resulted in synergistic effects.Conclusions:In ectopic endometrial stromal cells,there were aberrant methylation in the Promoter and exon region of PR-B,and the methylation closely related with PR-B expression.ADC and TSA can significantly inhibit the proliferation and block the cell cycle of ectopic endometrial stromal cells from women with adenomyoisis,and synergistically induce the expression of PR-B.