Investigation Of Integrin α₅ And Integrin β₃ In Endometriosis

Objectives: To study the expression and steroidal regulation of integrinα₅ and integrinβ₃ in eutopic endometrium and ectopic endometrium ,and to investigate the effects of estradiol,progesterone,anti- estradiol and anti-progesterone on the expression of integrinα₅,β₃mRNA in cultured human endometriotic cells in vitro.All of these purposes are to seach for the effeces of integrin on endometriosis,s occurrence and development , a:nd do our efforts for endometriosis,s diagnosis and treatment .Methods:1. Cultured human endometriotic cells in vitro: (1)Ectopic endometrial tissues were digested by enzymological technics and cultured in vitro.The cultured cells were dentified by mouse-anti-human keratin antibody and analyzed its purity. (2) The effects of estradiol(E₂),progesterone (P),tamoxifen(TAM) and mifepriston(RU486) on endometriotic cells'activity was detected by MTT. (3)After adhering to each other, all cultured ectopic endometrial cells were divided into five groups: control group,E₂ group,P group,TAM group and RU486 group.They were continue cultured in the same condition. (4)Estradiol(E₂,10nmol/L),progesterone(P,100nmol/L),tamoxifen(TAM,1000nmol/L) and mifepriston(RU486,1000nmol/L) acted on the cultured ectopic endometrial cells respectively for 5-7 days. The expression of integrinα₅,β₃ mRNA of each group were determined by reverse transcript polymerase chain reaction (RT-PCR) method. 2.Clinic part: (1) RT-PCR method was utilized to detect and quantitate expression levels of integrinα₅,β₃ mRNA in eutopic and ectopic endometrial tissues from 18 patients with endometriosis (study group) and 18 non-endometriosis women (control group) . (2) Serum concentrations of estradiol (E₂),progesterone(P) and CA125 were measured by radioimmunoassay. Every AFS-r of the state of pelvic cavity was scored during operation .Results: (1)The rate of successful culturing ectopic endometrial cells in vitro achieve 44.4%(8/18).(2) Estradiol(10nmol/L) and progesterone (100nmol/L) showed significantly effects on stimulating the growth of ectopic endometrial cells .Whereas,tamoxifen (1000nmol/L) and mifepriston (1000nmol/L) showed significantly effects on inhibiting the growth of ectopic endometrial cells. There was a certain relationship between drug's density and activity of endometrial cells. (3)The expression of integrinα₅ mRNA in E₂ and P group got 1.415±0.286 and 1.304±0.170 respectively ,which were significantly higher than that of control group(1.082±0.069),(P<0.05).In TAM and RU486 group,it was 0.882±0.013 and 0.866±0.054 respectively ,which were significantly lower than that of control group (P<0.05). The expression of integrinβ₃ mRNA in E₂ and P group was 0.844±0.003 and 0.803±0.005 ,which were significantly higher than that of control group(0.703±0.009),(P<0.05).In TAM and RU486 group,it was 0.604±0.003 and 0.592±0.011 respectively ,which were significantly lower than that of control group (P<0.05). (4) The various positive degrees of expression of integrin subunitsα₅ andβ₃ were shown in both eutopic and ectopic endometrial tissues of endometriosis and non-endometriosis.The expressions of integrinα₅ mRNA in eutopic endometrium ,ectopic endometrium and endometrium of control group were 0.773±0.113,1.077±0.032 and 0.924±0.120 respectively ,there was significant difference between every two groups(P<0.05). Moreover the expression of integrinβ₃ mRNA in eutopic endometrium ,ectopic endometrium and endometrium of control group weres 0.653±0.080,0.963±0.132 and 0.763±0.090 respectively ,there was significant difference between every two groups(P<0.05).(5) Serum concentration of E₂ in study group was positive correlated with the expression integrinα₅(r=0.793,P<0.05) andβ₃(r=0.793,0.917,P<0.05). While serum concentration of P has no relevance to integrinα₅ andβ₃ .Both level of serum CA125 and score of AFS-r have positive correlation with integrinα₅ andβ₃ (P<0.05) Conclusion:Integrinα₅ andβ₃ have some effects on endometriosis,s occurrence and development. (1) Estradiol and progesterone can promote the growth of ectopic endometrial cells ,while on which tamoxifen and mifepriston act an inhibiton effect ,and there was an relationship between drug's density and activity of endometrial cells.(2) Estradiol and progesterone may regulate the expression of integrinα₅ andβ₃,Estradioland progesterone can enhance the expression of integrinα₅ andβ₃ mRNA in ectopic endometral cell, and promote the occurrence of adhesion leading to endometriosis. Tamoxifen and mifepriston decelerate the expression of integrinα₅ andβ₃ mRNA in ectopic endometral cell.(3) The high expression of integrinα₅ andβ₃ in ectopic endometrium might play an importment role in the development of endometriosis adherion .(4)Liking CA125, integrinα₅ andβ₃ may become the assistant dignosis index of endometriosis.