Micro-RNA30c negatively regulates REDD1 expression in human hematopoietic and osteoblast cells after gamma-irradiation.
Micro-RNA30c negatively regulates REDD1 expression in human hematopoietic and osteoblast cells after gamma-irradiation.
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DOI：
10.1371/journal.pone.0048700
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Xiao M
中科院分区：
文献类型：
--
作者：
Li XH;Ha CT;Fu D;Xiao M
我们最近证明了一种新的细胞应激反应基因REDD1保护人类胎儿成骨细胞系（hFOB）细胞免受γ辐射诱导的过早衰老。在这里，我们发现内源性REDD1在人造血祖细胞CD34+细胞中的水平非常低，无论辐射如何，但在分化的造血细胞（培养14天的CD34+细胞）中高度表达，以响应辐射，这可能与后者细胞的辐射耐受性有关。为了进一步了解辐射对不同细胞的损伤机制，我们利用CD34+和hFOB细胞辐照前后纯化的miRNA进行了microRNA （miRNA）阵列，并利用实时反转录(RT)-PCR验证了miRNA在辐射损伤细胞中的表达谱。结果表明，γ辐射下调了CD34+细胞中的16个mirna和hFOB细胞中的14个mirna。辐射诱导CD34+细胞中的15个mirna上调，hFOB细胞中的18个mirna上调。辐射诱导的miRNA表达谱在CD34+和hFOB细胞中完全不同。辐射上调CD34+细胞中的miRNA (miR)-30b、miR-30c和miR-30d，而抑制hFOB细胞中miR-30c的表达。由于miR-30的潜在靶点位于REDD1基因的3 '非翻译区（UTR），并且辐射调节了miR-30c在CD34+和hFOB细胞中的表达，因此我们使用miR-30c抑制剂和前体（pre-miR-30c）进一步探索了miR-30c对REDD1表达的影响。结果表明，转染pre-miR-30c可抑制培养14天的CD34+细胞和hFOB细胞中REDD1的表达，导致hFOB细胞死亡。相反，抑制miR-30c表达可显著增强CD34+细胞的克隆原性。我们的数据表明，CD34+和hFOB细胞在辐照后具有不同的miRNA表达模式，miR-30c在辐射诱导的细胞损伤中起关键作用，可能是通过调节REDD1的表达。
We recently demonstrated that a novel cell stress response gene REDD1 protects human fetal osteoblast cell line (hFOB) cells from γ-radiation-induced premature senescence. Here we show that levels of endogenous REDD1 are very low in human hematopoietic progenitor CD34+ cells regardless of radiation, but highly expressed in differentiated hematopoietic cells (14 day cultured CD34+ cells) in response to radiation, which might be associated with radiation tolerance of the latter cells. To further understand the mechanisms of radiation-induced damage in different cells, microRNA (miRNA)-arrays were performed using purified miRNAs from CD34+ and hFOB cells before and post-irradiation and real-time reverse transcription (RT)-PCR was used to validate the expression profiles of miRNAs in the radiation-damaged cells. The results indicate that γ-radiation downregulated 16 miRNAs in CD34+ cells and 14 in hFOB cells. Radiation-induced upregulation was observed for 15 miRNAs in CD34+ cells and 18 miRNAs in hFOB cells. The profiles of radiation-induced miRNA expression were completely different in CD34+ vs. hFOB cells. Radiation up-regulated miRNA (miR)-30b, miR-30c and miR-30d in CD34+ cells, whereas it inhibited miR-30c expression in hFOB cells. Since miR-30 has potential target sites located in the 3′untranslated region (UTR) of the REDD1 gene and radiation regulated miR-30c expression in both CD34+ and hFOB cells, we further explored the effects of miR-30c on REDD1 expression using miR-30c inhibitor and precursor (pre-miR-30c). The results show that pre-miR-30c transfection suppressed REDD1 expression in 14 day cultured CD34+ cells and hFOB cells and resulted in hFOB cell death. In contrast, inhibition of miR-30c expression significantly enhanced clonogenicity in CD34+ cells. Our data suggest that CD34+ and hFOB cells have different miRNA expression patterns after irradiation and miR-30c plays a key role in radiation-induced cell damage which might be through regulation of REDD1 expression.
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影响因子：
16
作者：
Leung AK;Sharp PA
通讯作者：
Sharp PA
影响因子：
3.6
作者：
Baskar R;Lee KA;Yeo R;Yeoh KW
通讯作者：
Yeoh KW
影响因子：
3.5
作者：
Pesole, G;Mignone, F;Liuni, S
通讯作者：
Liuni, S
影响因子：
5.2
作者：
Yen, Men-Luh;Chien, Chih-Cheng;Yen, B. Linju
通讯作者：
Yen, B. Linju
DOI：
10.3324/haematol.2010.026492
发表时间：
2010-12-01
期刊：
HAEMATOLOGICA-THE HEMATOLOGY JOURNAL
影响因子：
--
作者：
Li, Xiang Hong;Fu, Dadin;Xiao, Mang
通讯作者：
Xiao, Mang