Genistein promotion of osteogenic differentiation through BMP2/SMAD5/RUNX2 signaling.
Genistein promotion of osteogenic differentiation through BMP2/SMAD5/RUNX2 signaling.
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DOI：
10.7150/ijbs.7367
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发表时间：
2013
影响因子：
9.2
通讯作者：
Xiao Z
中科院分区：
文献类型：
--
作者：
Dai J;Li Y;Zhou H;Chen J;Chen M;Xiao Z
为探讨Genistein对人骨髓间充质干细胞（hBMSC）成骨分化过程中成骨相关基因表达谱的影响，将hBMSC在含Genistein（10-8 × 10-5 M）的成骨分化培养基中培养12 d。BrdU掺入法检测细胞增殖，碱性磷酸酶（ALP）活性检测hBMSC的成骨分化。Caspase 3/7活性检测细胞凋亡。使用GEArray Q系列人成骨基因阵列分析与对照组相比，染料木素处理的hBMSC培养物中的大规模基因表达。定量实时RT-PCR，小干扰RNA（siRNA），和蛋白质印迹分析被用来确认在五个代表性的转录的微阵列数据。Genistein（10-8 × 10-6 M）剂量和时间依赖性地增加细胞增殖和细胞ALP活性，但对细胞凋亡无明显影响。96基因芯片分析表明，22个基因表达上调超过2倍，7个基因表达下调至少1.5倍。骨形态发生蛋白（BMPs），小母亲对decapentaplegic同源物（SMADs），和Runt相关转录因子2（RUNX 2）的表达伴随着金雀异黄素治疗下增加，而胰岛素样生长因子2和抑制SMADs 6和7的表达显着下降。实时荧光定量RT-PCR结果与基因芯片分析结果具有相关性，并具有雌激素受体依赖性。特异性基因siRNA敲低进一步证实了Genistein对BMP 2、SMAD 5和RUNX 2蛋白表达的成骨作用。Genistein主要通过BMP依赖的SMADs和RUNX 2信号通路促进hBMSCs的成骨分化。
To investigate the effects of Genistein on the osteogenic related gene expression profiles during osteoblastic differentiation of human bone marrow mesenchymal stem cell (hBMSC) cultures, the hBMSCs were cultured under osteogenic differentiation medium with the addition of Genistein (10-8∼10-5 M) for 12 days. The cell proliferation was measured by BrdU incorporation, while the osteoblastic differentiation in hBMSC cultures was assessed by cellular alkaline phosphatase (ALP) activity. The cell apoptosis was determined by caspase 3/7 activation. GEArray Q series human osteogenesis gene array was used to analyze large-scale gene expression in Genistein-treated hBMSC cultures compared to the control group. Quantitative real-time RT-PCR, small interfering RNA (siRNA), and western blot analysis were used to confirm the microarray data in five representative transcripts. Genistein (10-8∼10-6 M) dose- and time-dependently increased cell proliferation and cellular ALP activity, but had no significant effect on cell apoptosis in hBMSC cultures. The 96-gene array analysis indicated that 22 genes were upregulated more than 2-fold and 7 genes were downregulated at least 1.5-fold. The expressions of bone morphogenetic proteins (BMPs), small mothers against decapentaplegic homologs (SMADs), and Runt-related transcription factor 2 (RUNX2) were concomitantly increased under Genistein treatment while insulin-like growth factor 2 and inhibitory SMADs 6 and 7 expressions were significantly decreased. The results of the real-time RT-PCR had a correlation with the results of microarray analysis and were estrogen-receptor dependent. Specific gene siRNAs knock-down further confirmed the osteogenic effects of Genistein on BMP2, SMAD5 and RUNX2 protein expression. Genistein enhanced osteogenic differentiation in cultured hBMSCs mainly through the BMP-dependent SMADs and RUNX2 signaling.
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DOI：
10.1016/j.bbrc.2004.11.059
发表时间：
2005-01-21
影响因子：
3.1
作者：
Abdallah, BM;Haack-Sorensen, M;Kassem, M
通讯作者：
Kassem, M
影响因子：
158.5
作者：
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通讯作者：
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影响因子：
4.2
作者：
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影响因子：
4.8
作者：
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通讯作者：
Bendik, I
影响因子：
15.9
作者：
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通讯作者：
Robey, PG