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目的探讨卵巢激素是否影响体外培养的子宫肌瘤平滑肌细胞中(Fibrillin-1,FBN-1)mR-NA和蛋白的表达。方法10份子宫肌瘤及同源正常子宫平滑肌组织标本在术中被采集,游离的子宫肌瘤平滑肌细胞及同源正常子宫平滑肌细胞在含有10%的胎牛血清DMEM培养基中培养。在细胞达到70%融合时继续用无血清培养基培养48h,然后加入递增剂量的17β雌二醇(17-βestrodioal,E2)(0、0.1、1、10ng/ml)或E210ng/ml联合加入递增剂量的孕激素(0、1、10、100ng/ml)继续培养48h。采用实时荧光定量PCR、Western blot及荧光免疫组织化学方法探查FBN-1 mRNA和蛋白在体外培养的子宫肌瘤平滑肌细胞中的表达。结果免疫荧光组织化学染色显示FBN-1在子宫肌瘤平滑细胞及正常子宫平滑肌细胞中均有表达;E2呈剂量依赖性增加子宫肌瘤平滑细胞中FBN-1 mRNA和蛋白的表达,尤其在E2浓度增加为10ng/ml时,与对照组相比差异显著(P<0.05,P<0.01);而在E2联合不同剂量的孕激素的情况下对子宫平滑肌瘤细胞FBN-1 mRNA和蛋白表达无影响。结论雌激素增加体外培养的子宫肌瘤平滑细胞中FBN-1 mRNA和蛋白的表达。
Objective To investigate whether ovarian hormones affect the expression of mR-NA and protein in fibroblast smooth muscle cells (Fibrillin-1, FBN-1) cultured in vitro. Methods 10 specimens of uterine leiomyoma and normal uterine smooth muscle were collected intraoperatively. The free uterine myoma smooth muscle cells and normal uterine smooth muscle cells were cultured in DMEM containing 10% fetal bovine serum. When the cells reached 70% confluence, continue to culture with serum-free medium for 48h, and then add increasing doses of 17β-estrodioal (E2) (0,0.1,1,10ng / ml) or E210ng / ml The dose of progestin (0, 1, 10, 100 ng / ml) was continued for 48 h. The expression of FBN-1 mRNA and protein in cultured uterine fibroids was detected by real-time fluorescence quantitative PCR, Western blot and fluorescence immunohistochemistry. Results Immunofluorescence staining showed that FBN-1 was expressed in smooth muscle cells of uterine fibroids and normal uterine smooth muscle cells. E2 increased the expression of FBN-1 mRNA and protein in smooth muscle cells of uterine fibroids in a dose-dependent manner, especially in E2 (P <0.05, P <0.01) when the concentration was increased to 10 ng / ml, while the expression of FBN-1 mRNA and protein in uterine leiomyoma cells was increased in combination with different doses of progesterone no effect. Conclusion Estrogen increases the expression of FBN-1 mRNA and protein in smooth muscle cells of uterine fibroids in vitro.