Selection of suitable reference genes for normalization of quantitative real-time PCR in cartilage tissue injury and repair in rabbits.

Selection of suitable reference genes for normalization of quantitative real-time PCR in cartilage tissue injury and repair in rabbits.
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在软骨组织损伤和兔子修复中选择合适的参考基因,以归一化实时PCR。

DOI:
10.3390/ijms131114344
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发表时间:
2012-11-06
影响因子:
5.6
通讯作者:
Liang DC
Liang DC
中科院分区:
生物学2区
文献类型:
--
作者:
Peng XX;Zhao RL;Song W;Chu HR;Li M;Song SY;Li GZ;Liang DC

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当通过定量实时RT-PCR(RT-qPCR)研究与软骨再生相关的基因的表达变化时,需要在软骨细胞发育的不同阶段具有高度稳定表达的参考基因,以准确地标准化基因表达。迄今为止,尚未发表评价常用内参基因在兔关节软骨中表达变化的报道。在这项研究中,在有或没有胰岛素样生长因子1(IGF-1)治疗的情况下,在兔子的关节软骨中制造缺陷,以创造不同的软骨细胞生存环境。评估候选内参基因甘油醛-3-磷酸脱氢酶(GAPDH)、18S核糖体RNA(18S rRNA)、亲环蛋白(CYP)、次黄嘌呤磷酸核糖基转移酶(HPRT1)和β-2-微球蛋白(B2M)的表达稳定性和强度。数据通过 geNorm 和 NormFinder 进行分析。 B2M 和 18S rRNA 被鉴定为适合兔软骨组织的参考基因。
When studying the altered expression of genes associated with cartilage regeneration by quantitative real-time RT-PCR (RT-qPCR), reference genes with highly stable expression during different stages of chondrocyte developmental are necessary to normalize gene expression accurately. Until now, no reports evaluating expression changes of commonly used reference genes in rabbit articular cartilage have been published. In this study, defects were made in rabbit articular cartilage, with or without insulin-like growth factor 1 (IGF-1) treatment, to create different chondrocyte living environments. The stability and intensity of the expressions of the candidate reference genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S Ribosomal RNA (18S rRNA), cyclophilin (CYP), hypoxanthine phosphoribosyl transferase (HPRT1), and β-2-microglobulin (B2M) were evaluated. The data were analyzed by geNorm and NormFinder. B2M and 18S rRNA were identified to be suitable reference genes for rabbit cartilage tissues.
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