骨癌痛大鼠背根神经节中信使RNA表达谱的转录组学分析  

Transcriptomic analysis of mRNA expression profiles in dorsal root ganglia of rats with cancer-induced bone pain

作　　者：林铭雪 吴淑燕 郑晓春[1] 黄风怡[1] Lin Mingxue;Wu Shuyan;Zheng Xiaochun;Huang Fengyi(Department of Anesthesiology,Fujian Provincial Hospital,Fuzhou,350001,China)

机构地区：[1]福建省立医院麻醉科,福州350001

出　　处：《创伤与急诊电子杂志》2022年第3期121-128,共8页Journal of Trauma and Emergency(Electronic Version)

基　　金：福建省卫生健康科技计划医学创新项目(2020CXB002)。

摘　　要：目的基于RNA测序结果分析骨癌痛大鼠背根神经节中信使RNA(messenger RNA,mRNA)表达差异以及相关信号通路的改变。方法根据随机数表法将12只150~180g雌性SD大鼠随机分为假手术组和骨癌痛组,每组6只。骨癌痛组大鼠胫骨骨髓腔内注射MRMT-1乳腺癌细胞建立骨癌痛模型,假手术组注射等体积的D-hank's液作为对照。用Von Frey纤维丝测量两组大鼠50%机械缩足反射阈值(mechanical withdrawal threshold,MWT),采用影像学检测骨组织破坏情况,苏木精-伊红染色检测骨组织病理学改变,定量聚合酶链反应(quantitative PCR,qPCR)检测电压门控钠通道亚型1.6(voltagegated sodium channels subtype 1.6,Nav1.6)mRNA的表达量。结果①与假手术组相比,骨癌痛组大鼠50%MWT显著下降(F=20.50,P=0.006)。以差异倍数大于1.5倍且错误发现率小于0.05为条件,RNA测序筛查到149个表达显著差异的mRNA(上调124个,下调25个),其中Nav1.6与假手术组相比显著上调(2^(2.4)倍)。京都基因和基因组数据库富集分析,发现丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路是主要富集到的通路之一(富集基因数12,P=0.00059)。②骨癌痛组较假手术组,大鼠背根神经节中Nav1.6 mRNA相对表达量(1.80±0.31)明显高于假手术组(1.00±0.10),差异有统计学意义(P=0.01)。结论SD大鼠胫骨注射MRMT-1乳腺癌细胞可导致骨癌痛发生,其背根神经节中Nav1.6上调与MAPK信号通路激活可能是骨癌痛的发生机制之一。Objective To analyze the mRNA expression in the dorsal root ganglia(DRG)of rats with cancer-induced bone pain(CIBP)and the changes of related signaling pathways based on the results of transcriptome sequencing.Method Twelve Sprague-Dawley(SD)rats were randomly divided into CIBP group(n=6)and sham group(n=6).The rat model of CIBP was established by intramedullary injection of MRMT-1 cells into the tibia.Von Frey fiber was used to measure the mechanical withdrawal threshold(MWT)of rats.Bone tissue damage was detected by X-ray.Hematoxylin-eosin staining was used to detect bone histopathological changes,and quantitative polymerase chain reaction(qPCR)was used to detect the expression of voltage-gated sodium channels subtype 1.6(Nav1.6).Result Compared with the sham group,the 50%MWT of the CIBP group was significantly decreased(F=20.50,P=0.006).We ranked significant genes by fold-change with a cutoff of 1.5 and false discovery rate of 0.05,and 124 up-regulated genes and 25 down-regulated genes were detected,among which Nav1.6 was significantly up-regulated.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that differentially expressed genes were enriched in mitogen-activated protein kinase(MAPK)signaling pathway.Compared with sham group,the expression of Nav1.6 mRNA in DRG of rats in CIBP group were significantly increased(1.00±0.10 vs 1.80±0.31,P=0.01).Conclusion Injection of MRMT-1 breast cancer cells into the tibia of SD rats can lead to CIBP.The up-regulation of Nav1.6 in the DRG and the activation of MAPK signaling pathway are implicated in driving CIBP.

关 键 词：骨癌痛 背根神经节 RNA测序 电压门控钠通道 丝裂原活化蛋白激酶信号通路 

分 类 号：R738.1[医药卫生—肿瘤]