雌激素上调体外培养的子宫肌瘤平滑细胞中原纤维蛋白-1的表达

Upregulated expression of Fibrillin-1 mRNA and protein by estrogen in cultured human uterine leiomyomata cells

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 the effect of estrogen on fibrillin- 1 （FBN - 1） mRNA and protein expression in cultured leiomyomata smooth muscle cells （LSMC）. Methods Isolated LSMC were cultured in DMEM supplemented with 10% FBS and 1% antibiotic solution. The monolayer cells at approximately 70% confluence will continue to he cultured in serum - free DMEM medium for another 48 h and then in serum - free medium without or with 17-β estradiol （E2） at 0. 1, 1, 10ng/ml or E2 （10 ng/ml） plus progesterone at 0, 1, 10 and 100 ng/ml respectively for 48h. FBN - 1 mRNA and protein expressions in LSMC and myometrium smooth muscle cells （MSMC） were examined by quantitative real time PCR, Western blot and lmmunofluorescence staining. Results FBN - 1 protein was detected in the cytoplasm both in LSMC and MSMC. E2 significantly increased FBN - 1 mRNA and proteins expressions in LSMC in a dose - dependent manner, hut not in MSMC. E2 combined wtih different doses of progesterone did not affect FBN - 1 expression. Conclusion Estrogen increased FBN - 1 mRNA and protein expression in LSMC in a dose - dependent manner.