抗肿瘤多肽9R-P201诱导下的肝癌HepG2细胞转录组测序分析

Transcriptome Sequencing Analysis of Hepatocellular Carcinoma HepG2 Cells Induced by Antitumor Peptide 9R-P201

旨在探索多肽9R-P201处理肝癌HepG2细胞后基因融合、单核苷酸多态性(Single nucleotide polymorphism,SNP)突变、可变剪接等事件,并分析差异表达基因所参与的生物学进程与信号通路,以期解析多肽9R-P201在转录组水平对肝癌细胞的调控。通过转录组测序检测9R-P201处理肝癌HepG2细胞前后基因差异表达情况,tophat-fusion软件检测基因融合,SAMTOOLS软件检测SNP位点,r MATS软件鉴定可变剪接,使用基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析方法对差异表达基因进行功能富集分析。结果共检测到可变剪接事件276个、SNP位点5 557个、基因融合事件45个;同时共得到显著差异表达基因403个,其中上调269个而下调134个,基因的功能富集分析结果显示差异表达基因显著富集细胞生长、迁移等肿瘤相关生物进程,并参与多条与癌症相关的信号通路。研究表明在9R-P201诱导HepG2细胞后,导致表达差异基因显著与肿瘤生物学进程和通路相关,并发生了大量可变剪接、SNP突变、基因融合等事件,这暗示着该多肽有望作为后续肝癌介入治疗潜在药物分子。

This wok aims to elucidate the regulation of 9R-P201 on hepatoma cells（HepG2）at transcriptome level by investigatingthe gene fusion,SNP（Single Nucleotide Polymorphism）mutation,alternative splicing and other events after 9R-P201 treating HepG2,and analyzing the biological processes and signaling pathways involved by differentially expressed genes. The expression differences of thegenes before and after the 9R-P201 treating HepG2 cell line were detected by transcriptome sequencing. Meanwhile,gene fusion,SNPs,and alternative splicing were identified by tophat-fusion,SAMTOOLS software,and r MATS respectively. Functional enrichment analysis ofdifferentially expressed genes were performed by GO（Gene Ontology）and KEGG（Kyoto Encyclopedia of Genes and Genomes）. As results,276 alternative splicing events,5 557 SNP sites,and 45 gene fusion events were detected in the transcriptome sequencing. In addition,403 differentially expressed genes including 269 up-regulated and 134 down-regulated were detected. Gene GO and KEGG enrichment analysisshowed that differentially expressed genes were significantly involved in the biological processes of cell growth,locomotion,as well as anumber of cancer-related signaling pathways. In conclusion,the study revealed that the differentially expressed genes resulted from 9R-P201 treating HepG2 were significantly correlated with the biological processes and signaling pathways related to cancer and hepatocellular carcinomaHepG2 cell line,and a large number of alternative splicing events,SNP mutations,gene fusion events after 9R-P201 inducement occurred,suggesting this peptide may be used as a potential drug for subsequent hepatocellular carcinoma interventional therapy.