IL-6和TNF-α模拟炎性微环境促进人BMSCs向肿瘤相关成纤维细胞分化

摘要

Objective To observe whether BMSCs differentiate into TAFs in inflammatory microenvironment simula-ted by IL-6 and TNF-α.Methods The experiment was divided into 9 groups:the BMSCs group,the 50 ng/mL, 100 ng/mL IL-6 intervention group, the 50 ng/mL,100 ng/mL TNF-α intervention group, the 50 ng/mL IL-6+50 ng/mL TNF-α intervention group,the 50 ng/mL IL-6+100 ng/mL TNF-α intervention group,the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group and the 100 ng/mL IL-6+100 ng/mL TNF-α intervention group; After continuous induction for 42 days, BMSCs cell morphology and cycle, TAFs-tagged α-SMA and FAP protein were examined by phase-contrast microscope,flow cytometry and Western blot method. Results Compared with the normal control group, the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group BMSC cell proliferation was significantly promoted;G1phase decreased in proportion and S phase increased;α-SMA and FAP protein expression was signifi-cantly increased (P＜0.05).Conclusions Microenvironment simulated by the 100 ng/mL IL-6+50 ng/mL TNF-α may induce the abnormal change of the BMSCs morphological and proliferative characteristics. TAFs reference mole-cule α-SMA and FAP expression is increased.%目的 观察在IL-6和TNF-α模拟的炎性微环境中骨髓间充质干细胞( BMSCs)是否向肿瘤相关成纤维细胞(TAFs)分化.方法 将实验分为9组:对照组、IL-6 50 ng/mL及100 ng/mL干预组、TNF-α 50 ng/mL及100 ng/mL干预组、IL-6 50 ng/mL+TNF-α 50 ng/mL干预组、IL-6 50 ng/mL+TNF-α 100 ng/mL干预组、IL-6 100 ng/mL+TNF-α 100 ng/mL干预组、IL-6 100 ng/mL+TNF-α 50 ng/mL干预组;连续诱导42 d后,通过倒置相差显微镜、流式细胞计量术及Western blot分别检测BMSCs细胞形态、细胞周期及( TAFs)标记分子α-平滑肌肌动蛋白( α-SMA)、成纤维细胞活化蛋白(FAP)蛋白的表达.结果 与对照组相比,IL-6 100 ng/mL+TNF-α 50 ng/mL组可明显促进BMSCs细胞增殖;G1期比例降低,S 期比例升高,且 α-SMA,FAP 蛋白表达显著增加(P＜0.05).结论 IL-6 100 ng/mL+TNF-α 50 ng/mL所模拟的炎性微环境可诱导BMSCs细胞形态和增殖特性发生异常改变,TAFs标记分子α-SMA和FAP表达量升高.