插头框旋转录因子C2转染BMSCs后成骨作用的研究

发布时间：2018-11-17 20:19

【摘要】：[目的]观察过表达Foxc2基因对兔骨髓间充质干细胞(BMSCs)增殖及成骨分化的影响。[方法]构建携带Foxc2和绿色荧光蛋白(GFP)基因的重组慢病毒表达载体,分离、培养兔BMSCs,分别以Lenti-GFP(GFP组)、Lenti-Foxc2(Foxc2组)转染BMSCs,另设未转染BMSCs对照组。MTT法检测过表达Foxc2对细胞增殖能力;Western blot、Real time PCR检测转染后7d各组细胞Foxc2蛋白、m RNA表达;以及转染后1、2、4周各组细胞骨钙蛋白(OCN)、骨桥蛋白(OPN)、一型胶原(COLⅠ)蛋白及m RNA表达;转染后2周,各组行碱性磷酸酶(ALP)染色和ALP活性检测。[结果]Foxc2组Foxc2 m RNA和蛋白高效表达;对照组、GFP组无Foxc2 m RNA及蛋白表达;MTT法检测示过表达Foxc2对BMSCs的增殖未产生显著影响;Foxc2组OCN、OPN、COLⅠm RNA和蛋白表达显著高于其他组(P0.01),余组比较差异无统计学意义(P0.05);Foxc2组ALP染色阳性区域大于其余两组;ALP活性显著高于其他各组(P0.01)。[结论]过表达Foxc2不影响细胞增殖,可持续表达基因产物,并可促进BMSCs向成骨细胞分化。
[Abstract]:[objective] to observe the effect of overexpression of Foxc2 gene on (BMSCs) proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs). [methods] Recombinant lentivirus expression vector carrying Foxc2 and green fluorescent protein (GFP) gene was constructed. Rabbit BMSCs, was isolated and transfected into BMSCs, with Lenti-GFP (GFP group) and Lenti-Foxc2 (Foxc2 group) respectively. In addition, untransfected BMSCs was used as control group. MTT assay was used to detect the ability of Foxc2 expression on cell proliferation. The expression of Foxc2 protein, m RNA and osteocalcin (OCN), (OCN), osteopontin type I collagen (COL 鈪，

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