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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。
Objective: To observe the proliferation of uterine leiomyoma cells activated by stromal fibroblasts and to explore the role of stromal fibroblasts in the pathogenesis of uterine fibroids. Methods: Fibroblasts of uterine myoma and smooth muscle were sorted by immunomagnetic beads method. The expression of fibroblast activation marker (FAP) was detected by immunofluorescence and Western blot. The effect of fibroin-derived fibroblast culture supernatant on the expression of fibroblasts (Fib), smooth muscle cells (SMCs), fibroid cells UFC) activation and proliferation of vitality. RNAi technique was used to silence fibroblasts fibroblasts FAP gene, observed cell proliferation activity and ECM, growth factor expression changes, and observed cell proliferation pathway related signal molecules. Results: The FAP expression in fibroblasts of fibroids was higher than that in fibroblasts. The fibroblast fibroblast culture supernatant of fibroids could increase the FAP expression intensity of Fib and promote the cell proliferation activity. Silencing FAP gene could inhibit fibroblast stromal fibroblasts ECM (collgen I, fibronectin, laminin), growth factor (TGF-β, IGF-1) and cell proliferation pathway related signaling molecules (Akt, ERK, c-Fos, MEK) Conclusion: The activation of stromal fibroblasts in uterine fibroids is higher than that in uterine smooth muscle cells. The activated stromal fibroblasts have the effect of promoting cell proliferation, suggesting that activated stromal fibroblasts may play an important role in the development of uterine fibroids. effect.