Big endothelin changes the cellular miRNA environment in TMOb osteoblasts and increases mineralization.
Big endothelin changes the cellular miRNA environment in TMOb osteoblasts and increases mineralization.
复制标题
DOI：
10.3109/03008207.2014.923866
复制
发表时间：
2014-08
影响因子：
2.9
通讯作者：
Blank R
中科院分区：
文献类型：
--
作者：
Johnson MG;Kristianto J;Yuan B;Konicke K;Blank R
内皮素1（ET 1）促进成骨细胞乳腺癌和前列腺癌转移的生长。大ET 1转化为成熟ET 1，主要由内皮素转化酶1（ECE 1）催化，是ET 1生物活性所必需的。我们以前确定的Ece 1，基因座作为一个位置候选基因的多效性数量性状基因座影响股骨的大小，形状，矿化，和生物力学性能。我们将TMOb成骨细胞连续暴露于25 ng/ml大ET 1。细胞在生长培养基中生长6天，然后转换到含有或不含大ET 1的矿化培养基中，再生长15天，此时TMOb细胞形成矿化结节。我们通过茜素红染色定量矿化，并分析已知影响骨生成的miRNAs水平。微RNA（miR）126- 3 p通过搜索被鉴定为硬化蛋白（SOST）翻译的潜在调节因子。暴露于大ET 1的TMOb细胞显示出比对照细胞更大的矿化。大ET 1抑制靶向成骨蛋白转录物的miRNA。大ET 1增加靶向抑制骨生成的蛋白质转录物的miRNA的表达。与对照相比，大ET 1使126- 3 p的表达增加121倍。为了开始评估大ET 1对SOST产生的影响，我们分析了存在和不存在大ET 1的SOST转录和蛋白质产生，证明转录和翻译是解偶联的。我们的数据表明，大ET 1信号促进矿化。此外，结果表明，大ET 1的成骨作用可能是通过miR表达的变化介导的，这是一种以前未被认识到的大ET 1成骨机制。
Endothelin 1 (ET1) promotes the growth of osteoblastic breast and prostate cancer metastases. Conversion of big ET1 to mature ET1, catalyzed primarily by endothelin converting enzyme 1 (ECE1), is necessary for ET1’s biological activity. We previously identified the Ece1, locus as a positional candidate gene for a pleiotropic quantitative trait locus affecting femoral size, shape, mineralization, and biomechanical performance. We exposed TMOb osteoblasts continuously to 25 ng/ml big ET1. Cells were grown for 6 days in growth medium and then switched to mineralization medium with or without big ET1, for an additional 15 days by which time the TMOb cells form mineralized nodules. We quantified mineralization by alizarin red staining and analyzed levels of miRNAs known to affect osteogenesis. Micro RNA (miR) 126-3p was identified by search as a potential regulator of sclerostin (SOST) translation. TMOb cells exposed to big ET1 showed greater mineralization than control cells. Big ET1 repressed miRNAs targeting transcripts of osteogenic proteins. Big ET1 increased expression of miRNAs that target transcripts of proteins that inhibit osteogenesis. Big ET1 increased expression of 126-3p 121-fold versus control. To begin to assess the effect of big ET1 on SOST production we analyzed both SOST transcription and protein production with and without the presence of big ET1, demonstrating that transcription and translation were uncoupled. Our data show that big ET1 signaling promotes mineralization. Moreover, the results suggest that big ET1’s osteogenic effects are potentially mediated through changes in miR expression, a previously unrecognized big ET1 osteogenic mechanism.