Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer
Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer
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DOI：
10.1002/jbm.a.31130
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发表时间：
2007-07-01
影响因子：
4.9
通讯作者：
Hayashi, Yoshihiko
中科院分区：
文献类型：
--
作者：
Ganno, Tomoko;Yamada, Shizuka;Hayashi, Yoshihiko
壳聚糖具有多种生物活性。尽管有报道称壳聚糖促进骨病变的成骨，但对其如何在基因水平上调节硬组织形成细胞知之甚少。本研究主要研究了超低浓度壳聚糖单体培养成骨细胞的基因表达。从分离的RNA合成cDNA探针，并用荧光染料标记。将其与Human 3.8 II cDNA芯片杂交，分析荧光信号。cDNA微阵列分析显示，与对照组相比，3 d后，实验组中10个与各种信号相关分子相关的基因的表达水平>= 2.0倍。Real-time PCR分析显示，壳聚糖单体诱导了4个信号转导基因MAPK K3、MAPKKK11、Rac1和Shc1以及碱性磷酸酶基因的表达增加。这些结果表明，超低浓度的壳聚糖单体可以通过mRNA水平调节成骨细胞的活性，并直接影响细胞内的信号转导。(c) 2007 Wiley期刊公司
Chitosan has a variety of biological activities. Although it has been reported that chitosan promotes osteogenesis in bone lesions, little is known about how it modulates the hard tissue forming cells at the gene level. This study focused on gene expressions in osteoblasts cultured with a super-low concentration of chitosan monomer. cDNA probes were synthesized from isolated RNA and labeled with fluorescent dye. They were hybridized with Human 3.8 II cDNA microarray, and the fluorescent signal was analyzed. cDNA microarray analysis revealed that 10 genes concerning to various signaling-related molecules were expressed at >= 2.0-fold higher signal ratio levels in the experimental group when compared with the control group after 3 days. Real-time PCR analysis showed that chitosan monomer induced an increase in the expression of four signal transduction genes, mitogen-activated protein kinase (MAPK)K3, MAPKKK11, Rac1 and Shc1, together with the alkaline phosphatase gene. These results suggest that a super-low concentration of chitosan monomer could modulate the activity of osteoblastic cells through mRNA levels and that chitosan monomer directly affects signal transduction inside cells. (c) 2007 Wiley Periodicals, Inc.