摘要
背景与目的:外泌体是包含了复杂RNA和蛋白质的小膜泡,其中肿瘤细胞分泌的外泌体中的mRNA携带了大量肿瘤细胞遗传信息,因此对外泌体中特异性mRNA的分析,有望找到肿瘤诊断的新型分子标志物和治疗靶点。本研究通过高通量筛选与生物信息学方法探讨肝细胞癌(HCC)患者血清外泌体mRNA的表达特征及其潜在功能。方法:采集3例HCC患者及3例正常人的静脉血,用外泌体提取试剂盒提取血清外泌体,用Magen试剂盒提取外泌体RNA,对血清外泌体mRNA进行纯化、行反转录形成cDNA、PCR扩增、测序,最后将对所得数据质量进行评价后与参考数据进行比对,分析得到差异表达外泌体mRNA,并采用GO和KEGG通路富集分析对差异基因进行功能与通路注释。结果:与正常人比较,HCC患者有397个外泌体mRNA表达上调,192个外泌体mRNA表达下调;其中,NRGN、PF4、RGS18等17个基因表达明显上调,CXCL8、MORF4L2、SYCP1等14个基因显明显下调。GO富集分析显示,表达上调的外泌体mRNA的靶基因与蛋白质结合、蛋白质异二聚化活性、免疫系统过程的调节、胞外囊泡、细胞外细胞器、应激反应等有关,表达下调的外泌体mRNA的靶基因与嗅觉受体活性、细胞因子活性、CXCR趋化因子受体结合、中间丝、中间丝状细胞骨架等有关;KEGG通路分析显示,在上调的外泌体mRNA中,30条通路被显著富集,在下调的外泌体mRNA中9条通路被显著富集,其中,在上调的外泌体mRNA中,血小板激活、Rap 1信号通路、吞噬体、病毒致癌、肌动蛋白细胞骨架的调节与抗原处理和呈递等是最丰富和最有意义的通路;在下调的外泌体mRNA中,基础转录因子和细胞因子-细胞因子受体相互作用分别是最丰富和最有意义的通路。结论:HCC患者与正常人的血清外泌体mRNA表达特征存在较大差异,其与HCC发生、发展、转移可能密切相关,这为寻找新的诊断标志物及治疗靶点提
Background and Aims:Exosomes are small vesicles containing diverse RNAs and proteins,and those secreted from tumor cells carry considerable genetic information of the tumor cells. So, investigations of the specific mRNAs in the exosomes may provide chances for finding new molecular markers and therapeutic targets of tumors. This study was undertaken to investigate the expression profiles of the exosomal RNAs in hepatocellular carcinoma (HCC) patients and their potential functions by high-throughput screening and bioinformatics method. Methods: The venous blood samples were collected from 3 HCC patients and 3 healthy subjects, the serum exosomes were extracted by using exosome isolation kit, and exosomal RNAs were extracted by Magen kit. Then, the serum exosomal mRNAs were purified, reverse-transcribed into cDNAs, amplified by PCR, and identified by sequencing. Finally, the obtained data were compared with the reference data after quality assessment (BAM files) and the differentially expressed exosomal mRNAs were identified and evaluated, and the GO and KEGG Pathway enrichment analysis were used to annotate the function and pathway of the differentially expressed genes.Results: In HCC patients compared with healthy subjects, there were 397 up-regulated exosomal mRNAs and 192 down-regulated was up-regulated exosomal mRNAs, in which 17 genes such as NRGN, PF4 and RGS18 were significantly up-regulated, and 14 genes such as CXCL8, MORF4L2 and SYCP1 were significantly downregulated. GO enrichment analysis showed that the target genes of the up-regulated exosomal mRNAs were related to the protein binding, protein heterodimerization activity, regulation of the immune system process of adjusting and second extracellular vesicles, outside the cell organelles, stress reaction and so on, while the target genes of down-regulated exosomal mRNAs were related to olfactory receptor activity, cytokine activity, CXCR chemokine receptor binding, intermediate filaments, intermediate filamentous cytoskeleton. KEGG pathway analysis sh
作者
杨佳兴
徐红伟
王迎
王亮
王建国
李世朋
YANG Jiaxing;XU Hongwei;WANG Ying;WANG Liang;WANG Jianguo;LI Shipeng(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang,Henan 453100,China)
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2020年第1期53-60,共8页
China Journal of General Surgery
基金
国家自然科学基金资助项目(81700556)
河南省医学科技攻关基金资助项目(2018020358)
关键词
癌
肝细胞
外泌体
RNA
信使
生物标记
肿瘤
Carcinoma
Hepatocellular
Exosomes
RNA
Messenger
Biomarkers
Tumor
