骨母特异性因子2促进瘢痕疙瘩间充质干细胞体外增殖的研究

发布时间：2018-03-20 00:05

本文选题：瘢痕疙瘩　切入点：间充质干细胞　出处：《第三军医大学学报》2017年03期 　论文类型：期刊论文

【摘要】：目的探讨骨母特异性因子2对瘢痕疙瘩间充质干细胞(keloid mesenchymal stem cells,KMLSCs)体外增殖的影响。方法用瘢痕疙瘩和同体正常皮肤分离培养KMLSCs和皮肤干细胞(skin precursors,SKPs),传至第3代,流式细胞仪分选目的细胞并扩增。细胞免疫化学和Western blot检测KMLSCs和SKPs中骨母特异性因子2的表达;构建骨母特异性因子2慢病毒干扰表达载体并感染KMLSCs(干扰组、空载体和未转染组),利用qPCR和Western blot检测骨母特异性因子2 m RNA和蛋白水平的表达,MTT、流式细胞术(FCM)评价细胞增殖、凋亡和细胞周期,细胞免疫荧光和Western blot检测LRP5的表达。结果瘢痕疙瘩和皮肤中均可分离出干细胞,比例超过94%;免疫细胞化学和Western blot检测结果显示,KMLSCs中骨母特异性因子2的表达明显高于SKPs,且差异具有统计学意义(P0.05);qPCR和Western blot检测结果显示,骨母特异性因子2基因表达受明显干扰,MTT示干扰后KMLSCs增殖能力明显下降,FCM表明凋亡增强,细胞G1期阻滞;同时,细胞免疫与Western blot检测结果显示抑制骨母特异性因子2使LRP5表达明显减弱(P0.05)。结论沉默骨母特异性因子2的表达可诱导KMLSCs凋亡、阻滞细胞周期进展而使增殖能力降低,同时骨母特异性因子2引起KMLSCs增殖变化可能与LRP5改变有关。
[Abstract]:Objective to investigate the effect of osteoblastin-specific factor 2 on the proliferation of keloid mesenchymal stem cells in vitro. Methods KMLSCs and skin stem cells skin precursors were isolated from keloid and normal skin in vitro and transferred to the third generation. Cell immunocytochemistry and Western blot were used to detect the expression of osteoblast-specific factor 2 in KMLSCs and SKPs. QPCR and Western blot were used to detect the expression of bone mother specific factor 2 m RNA and protein. Flow cytometry was used to evaluate cell proliferation, apoptosis and cell cycle. Cell immunofluorescence and Western blot were used to detect the expression of LRP5. Results Stem cells were isolated from keloid and skin. The results of immunocytochemistry and Western blot showed that the expression of osteoblast specific factor 2 was significantly higher than that of SKPs2, and the difference was statistically significant (P 0.05) and Western blot. The expression of osteoblast-specific factor 2 gene was significantly interfered by MTT assay. The proliferation ability of KMLSCs was significantly decreased. FCM showed that apoptosis was enhanced and G1 phase was blocked, at the same time, the expression of osteoblast-specific factor 2 gene was inhibited. The results of cellular immunity and Western blot detection showed that inhibition of osteoblast-specific factor 2 significantly decreased the expression of LRP5. Conclusion silencing the expression of osteoblast-specific factor 2 can induce apoptosis of KMLSCs, block the progression of cell cycle and decrease the ability of proliferation. At the same time, the change of KMLSCs proliferation induced by bone mother specific factor 2 may be related to the change of LRP5.
【作者单位】：第三军医大学西南医院整形美容外科;
【基金】：国家自然科学基金面上项目(81272102)~~
【分类号】：R622

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