Discovery of Novel Cell Proliferation-Enhancing Gene by Random siRNA Library Based Combinatorial Screening
Discovery of Novel Cell Proliferation-Enhancing Gene by Random siRNA Library Based Combinatorial Screening
复制标题
通过基于随机 siRNA 文库的组合筛选发现新的细胞增殖增强基因。
DOI：
10.2174/138620710792927420
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发表时间：
2010-11-01
影响因子：
1.8
通讯作者：
Chen, Meihong
中科院分区：
文献类型：
--
作者：
Xiong, Xiahui;Lu, Yabin;Chen, Meihong
骨重塑受成骨细胞和破骨细胞的作用控制。成骨细胞增殖和分化受损可能破坏这种平衡，导致骨质疏松等病理症状。为了帮助理解成骨细胞增殖的分子机制，我们使用随机siRNA文库进行了表型驱动的高通量筛选，以寻找能够加速小鼠成骨前细胞MC3T3-E1增殖的新基因。从文库中筛选的三个sirna能够显著增强MC3T3-E1细胞的增殖。其中一种增殖增强sirna （B7）进一步进行了表达谱分析，以确定可能在其下游起作用的基因。许多基因受到增殖增强siRNA B7的调控。在这些基因中，尚未在细胞增殖中报道的Tmed2被证实过表达时能够增强MC3T3-E1细胞的增殖。我们的随机siRNA文库筛选过程提供了除了基因特异性siRNA文库之外的另一种策略，在基因组尺度上寻找新的功能基因。
Bone remodeling is tightly controlled by the actions of osteoblast and osteoclast. Impaired osteoblast proliferation and differentiation may disrupt the balance and lead to pathological symptom such as osteoporosis. To help understand the molecular mechanism of osteoblast proliferation, we performed a phenotype-driven high throughput screening with a random siRNA library, in search of novel genes that can accelerate murine preosteoblast MC3T3-E1 cell proliferation. Three siRNAs screened from the library were able to enhance MC3T3-E1 cell proliferation significantly. One of the proliferation-enhancing siRNAs (B7) was further subjected to expression profiling to pinpoint genes that putatively act down stream of it. A number of genes were regulated in response to proliferation-enhancing siRNA B7. Among these genes, Tmed2, which has never been reported yet in cell proliferation, was verified to be able to enhance MC3T3-E1 cell proliferation when over-expressed. Our screening process with random siRNA library provided an alternative strategy in addition to gene-specific siRNA library, in search of novel functional genes in genome scale.