miRNA基因在骨肉瘤患者外周血中的差异表达

Differential expression of microRNA in the serum of people with osteosarcoma

目的研究微小核糖核酸(microRNA,miRNA)基因在骨肉瘤患者外周血中的差异表达。方法收集郑州大学第一附属医院2013年3月至2015年4月收治的40例股骨远端和胫骨近端骨肉瘤患者,同期选择40例健康体检者。分别收集两组患者空腹静脉血,肿瘤组织及对照组相应部位组织,利用miRNA基因芯片对其进行分析,观察miRNA在外周血及骨肉瘤组织中的差异表达。结果筛查出骨肉瘤肿瘤组织中相关异常表达基因270个,上调miRNA基因15个,下调miRNA基因255个。上调及下调倍数大于2.5的miRNA 19个,上调大于2.5倍的miRNA基因8个:miR-21、miR-451、miR-218、miR-18a、miR-148a、miR-198、miR-195、miR-497。下调大于2.5倍11个:miR-29、miR-223、miR-183、miR-31、miR-654、miR-495、miR-493、miR-410、miR-409-5p、miR-409-3p、miR-382。外周血中异常表达基因178个,上调19个,下调159个。上调及下调倍数大于2.5的miRNA 21个,上调大于2.5倍的miRNA基因12个:miR-21、miR-451、miR-199a、miR-18a、miR-18b、miR-148a、miR-195、miR-497、miR-130b、miR-182、miR-374a、miR-301b。下调大于2.5倍的有9个:miR-29、miR-223、miR-410、miR-133b、miR-31、miR-206、miR-144、miR-493、miR-142-3p。两组结果有较高的重合性。其中表达特异性最明显的是miR-21(上调),miR-29(下调)。结论骨肉瘤肿瘤组织与外周血在miRNA表达上存在一定程度相关性和一致性,其中miR-21和miR-29特异性最明显,并预测其作为生物学标志物的可能性。

Objective To study the differential expression of microRNA（miRNA） gene in peripheral blood of patients with osteosarcoma,to establish the expression profile of miRNA gene in peripheral blood of osteosarcoma,and to analyze the up-regulation and down-regulation of miRNA gene. Methods Forty patients with distal femur or proximal tibia osteosarcoma of the First Affiliated Hospital of Zhengzhou University from March of 2013 to April of 2015 were enrolled in this study. There were 40 cases of venous blood and primary tumor tissue and 40 normal healthy people venous blood and corresponding normal tissue RNA. miRNAs were analyzed by gene chip. Results In the tissue of osteosarcoma,270 miRNA genes turned out to be differential（｜ Fold change ｜ ≥1. 74,P〈0. 05）. There were 15 up-regulated miRNA genes and 255 down-regulated miRNA genes,respectively. And 19 miRNA genes were expressed over 2. 5 fold ：.miR-21、miR-451、miR-218、miR-18 a、miR-148 a、miR-198、miR-195、miR-497 were expressed over 2. 5 fold higher than in healthy control subjects. miR-29、miR-223、miR-183、miR-31、miR-654、miR-495、miR-493、miR-410、miR-409-5 p、miR-409-3 p、miR-382 were expressed over 2. 5 fold lower than in healthy control subjects. In the serum of osteosarcoma,178 miRNA genes turned out to be differential（｜ Fold change ｜ ≥1. 74,P〈0. 05）. There were 19 up-regulated miRNA genes and 159 down-regulated miRNA genes,respectively. And 21 miRNA genes were expressed over 2. 5 fold. miR-21、miR-451、miR-199 a、miR-18 a、miR-18 b、miR-148 a、miR-195、miR-497、miR-130 b、miR-182、miR-374 a、miR-301 b were expressed over 2. 5 fold higher than in healthy control subjects. miR-29、miR-223、miR-410、miR-133 b、miR-31、miR-206、miR-144、miR-493、miR-142-3 p were expressed over 2. 5 fold lower than in healthy control subjects.The results of the two groups have a high coincidence. The most specific expressed genes were miR-21（up-regulated） and miR-29（down-regulated）. Conclusion The miRNA gene expression profile of peripheral blood of osteosarcoma was established,and it was proved that there was a certain degree of correlation and agreement between the tumor tissue and the peripheral blood of osteosarcoma. miR-21 and miR-29 were the most specific expressed genes,and predict the possibility of having it as a biological marker.