基质成纤维细胞活化促子宫肌瘤细胞增殖作用的研究

目的:观察基质成纤维细胞活化促进子宫肌瘤细胞的增殖作用,探讨基质成纤维细胞在子宫肌瘤发病机制中的作用。方法:采用免疫磁珠法分选子宫肌瘤组织和平滑肌组织的成纤维细胞。用免疫荧光及Western blot法检测成纤维细胞活化标记物(FAP)的表达;观察肌瘤基质成纤维细胞培养上清对平滑肌基质成纤维细胞(Fib)、平滑肌细胞(SMC)、肌瘤细胞(UFC)的活化作用及增殖活力的影响。采用RNAi技术沉默肌瘤基质成纤维细胞的FAP基因,观察细胞增殖活力及ECM、生长因子的表达变化,同时观察细胞增殖通路相关信号分子的改变。结果:肌瘤组织成纤维细胞中FAP表达高于Fib;肌瘤基质成纤维细胞培养上清能增加Fib的FAP表达强度,并促进细胞增殖活力;沉默FAP基因,可抑制肌瘤基质成纤维细胞ECM(collgen I、fibronectin、laminin)、生长因子(TGF-β、IGF-1)及细胞增殖通路相关信号分子(Akt、ERK、c-Fos、MEK)的表达。结论:子宫肌瘤基质成纤维细胞的活化程度高于子宫平滑肌组织基质成纤维细胞;活化的基质成纤维细胞具有促细胞增殖效应,提示活化的基质成纤维细胞在子宫肌瘤发生过程可能具有重要作用。

The effect of stromal fibroblasts activation on uterine leiomyoma cells proliferation and its mechanism

Objective: To observe the activation of stromal fibroblasts promotes the proliferation of uterine leiomyoma cells,and to investigate the role of stromal fibroblasts cells in the pathogenesis of uterine leiomyoma. Methods: Fibroblast cells from uterine leiomyoma and myometrium tissue was sorted by immunobead method. The expression of fibroblasts activation marker( FAP) was detected by immunofluorescence and Western blot. The activation and proliferation effect of uterine leiomyoma stromal fibroblasts cell culture supernatant on myometrium stromal fibroblasts( Fib) 、myometrium cells( SMC) and uterine leiomyoma cells( UFC) were tested. Using silence the FAP gene expression of leiomyoma stromal fibroblasts by RNA interference technology,and detect the cell proliferation ability and ECM、growth factor expression as well as the changing of signal molecule of cell proliferation pathway. Result: The FAP expression level of fibroblasts from leiomyoma tissue was higher than that from myometrium tissue. The cell culture supernatant of uterine leiomyoma stromal fibroblasts can enhance the FAP expression of Fib as well as the cell proliferation ability. Silence the expression of FAP gene can inhibit the expression of ECM( collgen I、fbronectin、laminin),TGF-β、IGF-1 and signal molecule of cell proliferation pathway( Akt、ERK、c-Fos、MEK). Conclusion: The activation level of leiomyoma stromal fibroblasts is higher than myometrium stromal fibroblasts. The activated fibroblast cells can enhance cell proliferation which prompt that the activation of fibroblast cells may play an important role in the pathogenesis of uterine leiomyoma.