本文選題：瘢痕疙瘩　切入點：間充質(zhì)干細胞　出處：《第三軍醫(yī)大學學報》2017年03期 　論文類型：期刊論文

【摘要】：目的探討骨母特異性因子2對瘢痕疙瘩間充質(zhì)干細胞(keloid mesenchymal stem cells,KMLSCs)體外增殖的影響。方法用瘢痕疙瘩和同體正常皮膚分離培養(yǎng)KMLSCs和皮膚干細胞(skin precursors,SKPs),傳至第3代,流式細胞儀分選目的細胞并擴增。細胞免疫化學和Western blot檢測KMLSCs和SKPs中骨母特異性因子2的表達;構(gòu)建骨母特異性因子2慢病毒干擾表達載體并感染KMLSCs(干擾組、空載體和未轉(zhuǎn)染組),利用qPCR和Western blot檢測骨母特異性因子2 m RNA和蛋白水平的表達,MTT、流式細胞術(shù)(FCM)評價細胞增殖、凋亡和細胞周期,細胞免疫熒光和Western blot檢測LRP5的表達。結(jié)果瘢痕疙瘩和皮膚中均可分離出干細胞,比例超過94%;免疫細胞化學和Western blot檢測結(jié)果顯示,KMLSCs中骨母特異性因子2的表達明顯高于SKPs,且差異具有統(tǒng)計學意義(P0.05);qPCR和Western blot檢測結(jié)果顯示,骨母特異性因子2基因表達受明顯干擾,MTT示干擾后KMLSCs增殖能力明顯下降,FCM表明凋亡增強,細胞G1期阻滯;同時,細胞免疫與Western blot檢測結(jié)果顯示抑制骨母特異性因子2使LRP5表達明顯減弱(P0.05)。結(jié)論沉默骨母特異性因子2的表達可誘導KMLSCs凋亡、阻滯細胞周期進展而使增殖能力降低,同時骨母特異性因子2引起KMLSCs增殖變化可能與LRP5改變有關(guān)。
[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.
【作者單位】： 第三軍醫(yī)大學西南醫(yī)院整形美容外科;
【基金】：國家自然科學基金面上項目(81272102)~~
【分類號】：R622

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