文摘
纤连蛋白(Fn)是一种细胞外基质糖蛋白,参与伤口修复,包括受伤的气道上皮的修复。支气管上皮细胞(bec)已知生产Fn活动相比,血清Fn具有增强的趋化作用。可变剪接的Fn基因是一个重要的细胞机制调节Fn的生产。产生Fn EIIIA地区包含人类bec,但IIICS地区的表达变异此前还没有报道。我们的目的是为了更好地定义人类的分子特征BEC Fn的表达通过确定替代拼接的变体IIICS地区Fn的BEC体外。人类支气管上皮细胞从支气管镜检查是有文化的。检查存在IIICS信使核糖核酸(mRNA)变异,我们合成的寡核苷酸引物互补发表人类纤连蛋白互补脱氧核糖核酸(互补)IIICS域的序列用于聚合酶链反应(PCR)总核糖核酸(RNA)提取培养人类bec。检查IIICS mRNA表达的调制,428碱基对(bp)产生的DNA片段PCR与32 oligo-labelled [P]脱氧胞苷三磷酸(dCTP)作为探针对北方污点分析。培养在人类bec的存在和缺乏转化生长因子(及)和代理影响环腺苷酸(cAMP)包括异丙肾上腺素和dibutryl营(db-cAMP)。从文化中提取总RNA,进行电泳,北部的屁股了。 Blots were hybridized with IIICS probe, total Fn cDNA, and tubulin cDNA. It was found that human BECs in culture expressed the five known human IIICS variants. TGF-beta enhanced the expression of IIICS mRNA in a concentration- and time-dependent fashion. Isoproterenol and db-cAMP both reduced the expression of IIICS mRNA and attenuated the TGF-beta induction. Changes in IIICS mRNA paralleled changes in total Fn mRNA, suggesting that these agents do not selectively modulate only the IIICS domain of Fn. We conclude that human airway epithelial cell Fn in vitro does contain mRNA for five IIICS variants, and that IIICS mRNA can be modulated by TGF-beta and agents which influence cAMP. It is unknown whether alterations in IIICS variants contribute to the functional differences previously observed between airway epithelial cell Fn and plasma-derived Fn.